Inhibition of colorectal cancer tumorigenesis by ursolic acid and doxorubicin is mediated by targeting the Akt signaling pathway and activating the Hippo signaling pathway

Hu,Dan; Meng,Ruo Yu; Nguyen,Thi Van; Chai,Ok Hee; Park,Byung Hyun; Lee,Ju-Seog; Kim,Soo Mi

doi:10.3892/mmr.2022.12898

Inhibition of colorectal cancer tumorigenesis by ursolic acid and doxorubicin is mediated by targeting the Akt signaling pathway and activating the Hippo signaling pathway

Authors:
- Dan Hu
- Ruo Yu Meng
- Thi Van Nguyen
- Ok Hee Chai
- Byung Hyun Park
- Ju-Seog Lee
- Soo Mi Kim
View Affiliations

Affiliations: Department of Physiology, Jeonbuk National University Medical School, Jeonju, Jeollabuk‑do 54907, Republic of Korea, Department of Physiology, Jeonbuk National University Medical School, Jeonju, Jeollabuk‑do 54907, Republic of Korea, Department of Anatomy, Jeonbuk National University Medical School, Jeonju, Jeollabuk‑do 54907, Republic of Korea, Department of Anatomy, Jeonbuk National University Medical School, Jeonju, Jeollabuk‑do 54907, Republic of Korea, Department of Biochemistry, Jeonbuk National University Medical School, Jeonju, Jeollabuk‑do 54907, Republic of Korea, Department of Systems Biology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
Published online on: November 15, 2022 https://doi.org/10.3892/mmr.2022.12898
Article Number: 11
Copyright: © Hu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Colorectal cancer (CRC) is one of the deadliest malignant tumors worldwide and its prevalence is increasing in South Korea. The efficacy of combined treatment with natural product‑derived and chemotherapy agents including curcumin combined with 5‑fluorouracil, resveratrol combined with cisplatin and epigallocatechin‑3‑gallate (EGCG) combined with cisplatin in preventing cancer progression and killing cancer cells has emerged. The Akt and Hippo signaling pathways serve a key role in colorectal tumor growth; however, the exact role of the crosstalk between Akt and Hippo signaling pathways in CRC remains poorly elucidated. The combined effect of UA and DOX on the cell proliferation, apoptosis, migration and cell cycle of CRC cells were investigated by performing Cell proliferation assay, a soft agar colony formation assay, flow cytometry, wound healing assay and western blotting assay. Subsequently, the expression of AKT and Hippo signaling pathway‑associated proteins were also assessed by western blot assay. Moreover, a xenograft nude mouse model was constructed to verify the effects of UA and DOX on the tumorigenesis of HCT116 cell in vivo. The present study reported that ursolic acid (UA) strongly enhanced the antitumor action of doxorubicin (DOX) via blocking the Akt/glycogen synthase kinase‑3β (Gsk3β) signaling pathway and activating tumor‑suppressive Hippo signaling (mammalian Ste20‑like kinase 1 and 2, salvador family WW domain containing protein 1 and MOB kinase activator 1), thereby downregulating downstream effector yes‑associated protein 1 (Yap) and connective tissue growth factor (CTGF) protein expression levels in CRC cells. Furthermore, The PI3K inhibitor LY294002 further suppressed Akt activity and enhance the function of Hippo pathway‑associated proteins in DOX + UA treated cells; this effect led to subsequent oncogenic Yap and CTGF inhibition following combined treatment, whereas Akt activator SC79 exerted an opposite effect in CTGF expression. In vivo, treatment with UA combined with DOX markedly suppressed the progression of CRC without any toxic effects on a xenograft mouse model by disrupting Akt signaling and activating the Hippo signaling pathway. These results demonstrated that UA and DOX treatment successfully induced Akt/Gsk3β inactivation via Hippo signaling pathway activation to promote Yap degradation, resulting in the inhibition of colorectal tumorigenesis. In conclusion, these findings suggested that combination therapy with UA and DOX may be more effective than DOX alone. UA may be a novel anticancer strategy and could be considered for investigation as a complementary chemotherapy agent in the future.

View Figures

View References

1	Arnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A and Bray F: Global patterns and trends in colorectal cancer incidence and mortality. Gut. 66:683–691. 2017. View Article : Google Scholar : PubMed/NCBI
2	Shin DW, Chang D, Jung JH, Han K, Kim SY, Choi KS, Lee WC and Park JH and Park JH: Disparities in the participation rate of colorectal cancer screening by fecal occult blood test among people with disabilities: A national database study in South Korea. Cancer Res Treat. 52:60–73. 2020. View Article : Google Scholar : PubMed/NCBI
3	Yang SY, Cho MS and Kim NK: Difference between right-sided and left-sided colorectal cancers: From embryology to molecular subtype. Expert Rev Anticancer Ther. 18:351–358. 2018. View Article : Google Scholar : PubMed/NCBI
4	McQuade RM, Stojanovska V, Bornstein JC and Nurgali K: Colorectal cancer chemotherapy: The evolution of treatment and new approaches. Curr Med Chem. 24:1537–1557. 2017. View Article : Google Scholar : PubMed/NCBI
5	Cargnin ST and Gnoatto SB: Ursolic acid from apple pomace and traditional plants: A valuable triterpenoid with functional properties. Food Chem. 220:477–489. 2017. View Article : Google Scholar : PubMed/NCBI
6	Xu C, Liao Y, Fang C, Tsunoda M, Zhang Y, Song Y and Deng S: Simultaneous analysis of ursolic acid and oleanolic acid in guava leaves using QuEChERS-based extraction followed by high-performance liquid chromatography. J Anal Methods Chem. 2017:29845622017. View Article : Google Scholar : PubMed/NCBI
7	Zheng JL, Wang SS, Shen KP, Huang XW, Li M, Chen L, Peng X, An HM and Hu B: Ursolic acid potentiated oxaliplatin to induce apoptosis in colorectal cancer RKO cells. Pharmazie. 75:246–249. 2020.PubMed/NCBI
8	Wang X, Wang T, Yi F, Duan C, Wang Q, He N, Zhu L, Li Q and Deng W: Ursolic acid inhibits tumor growth via epithelial-to-mesenchymal transition in colorectal cancer cells. Biol Pharm Bull. 42:685–691. 2019. View Article : Google Scholar : PubMed/NCBI
9	Cai Q, Lin J, Zhang L, Lin J, Wang L, Chen D and Peng J: Comparative proteomics-network analysis of proteins responsible for ursolic acid-induced cytotoxicity in colorectal cancer cells. Tumour Biol. 39:10104283176950152017. View Article : Google Scholar : PubMed/NCBI
10	Wang C, Shu L, Zhang C, Li W, Wu R, Guo Y, Yang Y and Kong AN: Histone methyltransferase Setd7 regulates Nrf2 signaling pathway by phenethyl isothiocyanate and ursolic acid in human prostate cancer cells. Mol Nutr Food Res. 62:e17008402018. View Article : Google Scholar : PubMed/NCBI
11	Yang K, Chen Y, Zhou J, Ma L, Shan Y, Cheng X, Wang Y, Zhang Z, Ji X, Chen L, et al: Ursolic acid promotes apoptosis and mediates transcriptional suppression of CT45A2 gene expression in non-small-cell lung carcinoma harbouring EGFR T790M mutations. Br J Pharmacol. 176:4609–4624. 2019. View Article : Google Scholar : PubMed/NCBI
12	Mendes VIS, Bartholomeusz GA, Ayres M, Gandhi V and Salvador JAR: Synthesis and cytotoxic activity of novel A-ring cleaved ursolic acid derivatives in human non-small cell lung cancer cells. Eur J Med Chem. 123:317–331. 2016. View Article : Google Scholar : PubMed/NCBI
13	Chan EWC, Soon CY, Tan JBL, Wong SK and Hui YW: Ursolic acid: An overview on its cytotoxic activities against breast and colorectal cancer cells. J Integr Med. 17:155–160. 2019. View Article : Google Scholar : PubMed/NCBI
14	Kim K, Shin EA, Jung JH, Park JE, Kim DS, Shim BS and Kim SH: Ursolic acid induces apoptosis in colorectal cancer cells partially via upregulation of MicroRNA-4500 and inhibition of JAK2/STAT3 phosphorylation. Int J Mol Sci. 20:1142018. View Article : Google Scholar : PubMed/NCBI
15	Prasad S, Yadav VR, Sung B, Reuter S, Kannappan R, Deorukhkar A, Diagaradjane P, Wei C, Baladandayuthapani V, Krishnan S, et al: Ursolic acid inhibits growth and metastasis of human colorectal cancer in an orthotopic nude mouse model by targeting multiple cell signaling pathways: Chemosensitization with capecitabine. Clin Cancer Res. 18:4942–4953. 2012. View Article : Google Scholar : PubMed/NCBI
16	Liu P, Du R and Yu X: Ursolic acid exhibits potent anticancer effects in human metastatic melanoma cancer cells (SK-MEL-24) via apoptosis induction, inhibition of cell migration and invasion, cell cycle arrest, and inhibition of mitogen-activated protein kinase (MAPK)/ERK signaling pathway. Med Sci Monit. 25:1283–1290. 2019. View Article : Google Scholar : PubMed/NCBI
17	Liu T, Ma H, Shi W, Duan J, Wang Y, Zhang C, Li C, Lin J, Li S, Lv J and Lin L: Inhibition of STAT3 signaling pathway by ursolic acid suppresses growth of hepatocellular carcinoma. Int J Oncol. 51:555–562. 2017. View Article : Google Scholar : PubMed/NCBI
18	Zhang L, Cai QY, Liu J, Peng J, Chen YQ, Sferra TJ and Lin JM: Ursolic acid suppresses the invasive potential of colorectal cancer cells by regulating the TGF-β1/ZEB1/miR-200c signaling pathway. Oncol Lett. 18:3274–3282. 2019.PubMed/NCBI
19	Cheng J, Liu Y, Liu Y, Liu D, Liu Y, Guo Y, Wu Z, Li H and Wang H: Ursolic acid alleviates lipid accumulation by activating the AMPK signaling pathway in vivo and in vitro. J Food Sci. 85:3998–4008. 2020. View Article : Google Scholar : PubMed/NCBI
20	Kim GH, Kan SY, Kang H, Lee S, Ko HM, Kim JH and Lim JH: Ursolic acid suppresses cholesterol biosynthesis and exerts anti-cancer effects in hepatocellular carcinoma cells. Int J Mol Sci. 20:47672019. View Article : Google Scholar : PubMed/NCBI
21	Lin CW, Chin HK, Lee SL, Chiu CF, Chung JG, Lin ZY, Wu CY, Liu YC, Hsiao YT, Feng CH, et al: Ursolic acid induces apoptosis and autophagy in oral cancer cells. Environ Toxicol. 34:983–991. 2019. View Article : Google Scholar : PubMed/NCBI
22	Lin JH, Chen SY, Lu CC, Lin JA and Yen GC: Ursolic acid promotes apoptosis, autophagy, and chemosensitivity in gemcitabine-resistant human pancreatic cancer cells. Phytother Res. 34:2053–2066. 2020. View Article : Google Scholar : PubMed/NCBI
23	Lin W and Ye H: Anticancer activity of ursolic acid on human ovarian cancer cells via ROS and MMP mediated apoptosis, cell cycle arrest and downregulation of PI3K/AKT pathway. J BUON. 25:750–756. 2020.PubMed/NCBI
24	Li W, Zhang H, Nie M, Tian Y, Chen X, Chen C, Chen H and Liu R: Ursolic acid derivative FZU-03,010 inhibits STAT3 and induces cell cycle arrest and apoptosis in renal and breast cancer cells. Acta Biochim Biophys Sin (Shanghai). 49:367–373. 2017. View Article : Google Scholar : PubMed/NCBI
25	Ruan JS, Zhou H, Yang L, Wang L, Jiang ZS, Sun H and Wang SM: Ursolic acid attenuates TGF-β1-induced epithelial-mesenchymal transition in NSCLC by targeting integrin αVβ5/MMPs signaling. Oncol Res. 27:593–600. 2019. View Article : Google Scholar : PubMed/NCBI
26	Sohn EJ, Won G, Lee J, Yoon SW, Lee I, Kim HJ and Kim SH: Blockage of epithelial to mesenchymal transition and upregulation of let 7b are critically involved in ursolic acid induced apoptosis in malignant mesothelioma cell. Int J Biol Sci. 12:1279–1288. 2016. View Article : Google Scholar : PubMed/NCBI
27	Lee NR, Meng RY, Rah SY, Jin H, Ray N, Kim SH, Park BH and Kim SM: Reactive oxygen species-mediated autophagy by ursolic acid inhibits growth and metastasis of esophageal cancer cells. Int J Mol Sci. 21:94092020. View Article : Google Scholar : PubMed/NCBI
28	Park HJ, Jo DS, Choi DS, Bae JE, Park NY, Kim JB, Chang JH, Shin JJ and Cho DH: Ursolic acid inhibits pigmentation by increasing melanosomal autophagy in B16F1 cells. Biochem Biophys Res Commun. 531:209–214. 2020. View Article : Google Scholar : PubMed/NCBI
29	Arcamone F, Cassinelli G, Fantini G, Grein A, Orezzi P, Pol C and Spalla C: Adriamycin, 14-hydroxydaunomycin, a new antitumor antibiotic from S. peucetius var. caesius. Reprinted from biotechnology and bioengineering, Vol. XI, Issue 6, Pages 1101–1110 (1969). Biotechnol Bioeng. 67:704–713. 2000. View Article : Google Scholar : PubMed/NCBI
30	Cortés-Funes H and Coronado C: Role of anthracyclines in the era of targeted therapy. Cardiovasc Toxicol. 7:56–60. 2007. View Article : Google Scholar : PubMed/NCBI
31	Weiss RB: The anthracyclines: Will we ever find a better doxorubicin? Semin Oncol. 19:670–686. 1992.PubMed/NCBI
32	Sarmento-Ribeiro AB, Scorilas A, Goncalves AC, Efferth T and Trougakos IP: The emergence of drug resistance to targeted cancer therapies: Clinical evidence. Drug Resist Updat. 47:1006462019. View Article : Google Scholar : PubMed/NCBI
33	Rui M, Xin Y, Li R, Ge Y, Feng C and Xu X: Targeted biomimetic nanoparticles for synergistic combination chemotherapy of paclitaxel and doxorubicin. Mol Pharm. 14:107–123. 2017. View Article : Google Scholar : PubMed/NCBI
34	Fan YP, Liao JZ, Lu YQ, Tian DA, Ye F, Zhao PX, Xiang GY, Tang WX and He XX: MiR-375 and doxorubicin co-delivered by liposomes for combination therapy of hepatocellular carcinoma. Mol Ther Nucleic Acids. 7:181–189. 2017. View Article : Google Scholar : PubMed/NCBI
35	Minotti G, Menna P, Salvatorelli E, Cairo G and Gianni L: Anthracyclines: Molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev. 56:185–229. 2004. View Article : Google Scholar : PubMed/NCBI
36	Guo NF, Cao YJ, Chen X, Zhang Y, Fan YP, Liu J and Chen XL: Lixisenatide protects doxorubicin-induced renal fibrosis by activating wNF-κB/TNF-α and TGF-β/Smad pathways. Eur Rev Med Pharmacol Sci. 23:4017–4026. 2019.PubMed/NCBI
37	Saleh D, Abdelbaset M, Hassan A, Sharaf O, Mahmoud S and Hegazy R: Omega-3 fatty acids ameliorate doxorubicin-induced cardiorenal toxicity: In-vivo regulation of oxidative stress, apoptosis and renal Nox4, and in-vitro preservation of the cytotoxic efficacy. PLoS One. 15:e02421752020. View Article : Google Scholar : PubMed/NCBI
38	Prasanna PL, Renu K and Valsala Gopalakrishnan A: New molecular and biochemical insights of doxorubicin-induced hepatotoxicity. Life Sci. 250:1175992020. View Article : Google Scholar : PubMed/NCBI
39	Zhou X, Xu P, Dang R, Guo Y, Li G, Qiao Y, Xie R, Liu Y and Jiang P: The involvement of autophagic flux in the development and recovery of doxorubicin-induced neurotoxicity. Free Radic Biol Med. 129:440–445. 2018. View Article : Google Scholar : PubMed/NCBI
40	Yu FX, Zhao B and Guan KL: Hippo pathway in organ size control, tissue homeostasis, and cancer. Cell. 163:811–828. 2015. View Article : Google Scholar : PubMed/NCBI
41	Kim CL, Choi SH and Mo JS: Role of the Hippo pathway in fibrosis and cancer. Cells. 8:4682019. View Article : Google Scholar : PubMed/NCBI
42	Dong J, Feldmann G, Huang J, Wu S, Zhang N, Comerford SA, Gayyed MF, Anders RA, Maitra A and Pan D: Elucidation of a universal size-control mechanism in Drosophila and mammals. Cell. 130:1120–1233. 2007. View Article : Google Scholar : PubMed/NCBI
43	Avruch J, Zhou D and Bardeesy N: YAP oncogene overexpression supercharges colon cancer proliferation. Cell Cycle. 11:1090–1096. 2012. View Article : Google Scholar : PubMed/NCBI
44	Liu XF, Han Q, Rong XZ, Yang M, Han YC, Yu JH and Lin XY: ANKHD1 promotes proliferation and invasion of non-small-cell lung cancer cells via regulating YAP oncoprotein expression and inactivating the Hippo pathway. Int J Oncol. 56:1175–1185. 2020.PubMed/NCBI
45	Niu K, Liu Y, Zhou Z, Wu X, Wang H and Yan J: Antitumor effects of paeoniflorin on Hippo signaling pathway in gastric cancer cells. J Oncol. 2021:47249382021. View Article : Google Scholar : PubMed/NCBI
46	Hou L, Chen L and Fang L: Scutellarin inhibits proliferation, invasion, and tumorigenicity in human breast cancer cells by regulating HIPPO-YAP signaling pathway. Med Sci Monit. 23:5130–5138. 2017. View Article : Google Scholar : PubMed/NCBI
47	Driskill JH and Pan D: The Hippo pathway in liver homeostasis and pathophysiology. Annu Rev Pathol. 16:299–322. 2021. View Article : Google Scholar : PubMed/NCBI
48	Masliantsev K, Karayan-Tapon L and Guichet PO: Hippo signaling pathway in gliomas. Cells. 10:1842021. View Article : Google Scholar : PubMed/NCBI
49	Ansari D, Ohlsson H, Althini C, Bauden M, Zhou Q, Hu D and Andersson R: The Hippo signaling pathway in pancreatic cancer. Anticancer Res. 39:3317–3321. 2019. View Article : Google Scholar : PubMed/NCBI
50	Llado V, Nakanishi Y, Duran A, Reina-Campos M, Shelton PM, Linares JF, Yajima T, Campos A, Aza-Blanc P, Leitges M, et al: Repression of intestinal stem cell function and tumorigenesis through direct phosphorylation of β-catenin and Yap by PKCζ. Cell Rep. 10:740–754. 2015. View Article : Google Scholar : PubMed/NCBI
51	Chen L, Qin F, Deng X, Avruch J and Zhou D: Hippo pathway in intestinal homeostasis and tumorigenesis. Protein Cell. 3:305–310. 2012. View Article : Google Scholar : PubMed/NCBI
52	Gu Y, Zhang L and Yu FX: Functions and regulations of the Hippo signaling pathway in intestinal homeostasis, regeneration and tumorigenesis. Yi Chuan. 39:588–596. 2017.PubMed/NCBI
53	Zhou D, Zhang Y, Wu H, Barry E, Yin Y, Lawrence E, Dawson D, Willis JE, Markowitz SD, Camargo FD and Avruch J: Mst1 and Mst2 protein kinases restrain intestinal stem cell proliferation and colonic tumorigenesis by inhibition of Yes-associated protein (Yap) overabundance. Proc Natl Acad Sci USA. 108:E1312–E1320. 2011. View Article : Google Scholar : PubMed/NCBI
54	Xiao Y, Liu Q, Peng N, Li Y, Qiu D, Yang T, Kang R, Usmani A, Amadasu E, Borlongan CV and Yu G: Lovastatin inhibits RhoA to suppress canonical Wnt/β-catenin signaling and alternative Wnt-YAP/TAZ signaling in colon cancer. Cell Transplant. 31:96368972210757492022. View Article : Google Scholar : PubMed/NCBI
55	Touil Y, Igoudjil W, Corvaisier M, Dessein AF, Vandomme J, Monté D, Stechly L, Skrypek N, Langlois C, Grard G, et al: Colon cancer cells escape 5FU chemotherapy-induced cell death by entering stemness and quiescence associated with the c-Yes/YAP axis. Clin Cancer Res. 20:837–846. 2014. View Article : Google Scholar : PubMed/NCBI
56	Shamekhi S, Abdolalizadeh J, Ostadrahimi A, Mohammadi SA, Barzegari A, Lotfi H, Bonabi E and Zarghami N: Apoptotic effect of saccharomyces cerevisiae on human colon cancer SW480 cells by regulation of Akt/NF-ĸB signaling pathway. Probiotics Antimicrob Proteins. 12:311–319. 2020. View Article : Google Scholar : PubMed/NCBI
57	Goel S, Huang J and Klampfer L: K-Ras, intestinal homeostasis and colon cancer. Curr Clin Pharmacol. 10:73–81. 2015. View Article : Google Scholar : PubMed/NCBI
58	Tumaneng K, Schlegelmilch K, Russell RC, Yimlamai D, Basnet H, Mahadevan N, Fitamant J, Bardeesy N, Camargo FD and Guan KL: YAP mediates crosstalk between the Hippo and PI(3)K-TOR pathways by suppressing PTEN via miR-29. Nat Cell Biol. 14:1322–1329. 2012. View Article : Google Scholar : PubMed/NCBI
59	Yu FX, Zhao B, Panupinthu N, Jewell JL, Lian I, Wang LH, Zhao J, Yuan H, Tumaneng K, Li H, et al: Regulation of the Hippo-YAP pathway by G-protein-coupled receptor signaling. Cell. 150:780–791. 2012. View Article : Google Scholar : PubMed/NCBI
60	Kawai K, Viars C, Arden K, Tarin D, Urquidi V and Goodison S: Comprehensive karyotyping of the HT-29 colon adenocarcinoma cell line. Genes Chromosomes Cancer. 34:1–8. 2002. View Article : Google Scholar : PubMed/NCBI
61	Grada A, Otero-Vinas M, Prieto-Castrillo F, Obagi Z and Falanga V: Research techniques made simple: Analysis of collective cell migration using the wound healing assay. J Invest Dermatol. 137:e11–e16. 2017. View Article : Google Scholar : PubMed/NCBI
62	Vang Mouritzen M and Jenssen H: Optimized scratch assay for in vitro testing of cell migration with an automated optical camera. J Vis Exp. 576912018.PubMed/NCBI
63	Martinotti S and Ranzato E: Scratch wound healing assay. Methods Mol Biol. 2109:225–229. 2020. View Article : Google Scholar : PubMed/NCBI
64	Meng RY, Jin H, Nguyen TV, Chai OH, Park BH and Kim SM: Ursolic acid accelerates paclitaxel-induced cell death in esophageal cancer cells by suppressing Akt/FOXM1 signaling cascade. Int J Mol Sci. 22:114862021. View Article : Google Scholar : PubMed/NCBI
65	Stephenson W: Deficiencies in the national institute of health's guidelines for the care and protection of laboratory animals. J Med Philos. 18:375–88. 1993. View Article : Google Scholar : PubMed/NCBI
66	Kilkenny C, Browne WJ, Cuthill IC, Emerson M and Altman DG: Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. Osteoarthritis Cartilage. 20:256–260. 2012. View Article : Google Scholar : PubMed/NCBI
67	Spangenberg EM and Keeling LJ: Assessing the welfare of laboratory mice in their home environment using animal-based measures-a benchmarking tool. Lab Anim. 50:30–38. 2016. View Article : Google Scholar : PubMed/NCBI
68	Clayden EC: Practical section cutting and staining. 5th edition. Edinburgh: (15 Teviot Place, Edinburgh 1). Churchill Livingstone; 7. pp. pp2701971
69	Kim DH, Kang DY, Sp N, Jo ES, Rugamba A, Jang KJ and Yang YM: Methylsulfonylmethane induces cell cycle arrest and apoptosis, and suppresses the stemness potential of HT-29 cells. Anticancer Res. 40:5191–5200. 2020. View Article : Google Scholar : PubMed/NCBI
70	Ombrato L and Malanchi I: The EMT universe: Space between cancer cell dissemination and metastasis initiation. Crit Rev Oncog. 19:349–361. 2014. View Article : Google Scholar : PubMed/NCBI
71	Wang C, Li Z, Lu Y, Du R, Katiyar S, Yang J, Fu M, Leader JE, Quong A, Novikoff PM and Pestell RG: Cyclin D1 repression of nuclear respiratory factor 1 integrates nuclear DNA synthesis and mitochondrial function. Proc Natl Acad Sci USA. 103:11567–11572. 2006. View Article : Google Scholar : PubMed/NCBI
72	Bishnupuri KS, Alvarado DM, Khouri AN, Shabsovich M, Chen B, Dieckgraefe BK and Ciorba MA: IDO1 and kynurenine pathway metabolites activate PI3K-Akt signaling in the neoplastic colon epithelium to promote cancer cell proliferation and inhibit apoptosis. Cancer Res. 79:1138–1150. 2019. View Article : Google Scholar : PubMed/NCBI
73	Ji J, Wang Z, Sun W, Li Z, Cai H, Zhao E and Cui H: Effects of cynaroside on cell proliferation, apoptosis, migration and invasion though the MET/AKT/mTOR axis in gastric cancer. Int J Mol Sci. 22:121252021. View Article : Google Scholar : PubMed/NCBI
74	Zhang P, Yuan X, Yu T, Huang H, Yang C, Zhang L, Yang S, Luo X and Luo J: Lycorine inhibits cell proliferation, migration and invasion, and primarily exerts in vitro cytostatic effects in human colorectal cancer via activating the ROS/p38 and AKT signaling pathways. Oncol Rep. 45:192021. View Article : Google Scholar : PubMed/NCBI
75	Yang L, Liu Y, Wang M, Qian Y, Dong X, Gu H, Wang H, Guo S and Hisamitsu T: Quercetin-induced apoptosis of HT-29 colon cancer cells via inhibition of the Akt-CSN6-Myc signaling axis. Mol Med Rep. 14:4559–4566. 2016. View Article : Google Scholar : PubMed/NCBI
76	Guo C, Zhang X and Pfeifer GP: The tumor suppressor RASSF1A prevents dephosphorylation of the mammalian STE20-like kinases MST1 and MST2. J Biol Chem. 286:6253–6261. 2011. View Article : Google Scholar : PubMed/NCBI
77	Kim M, Kim M, Lee MS, Kim CH and Lim DS: The MST1/2-SAV1 complex of the Hippo pathway promotes ciliogenesis. Nat Commun. 5:53702014. View Article : Google Scholar : PubMed/NCBI
78	Shome D, von Woedtke T, Riedel K and Masur K: The HIPPO transducer YAP and its targets CTGF and Cyr61 drive a paracrine signalling in cold atmospheric plasma-mediated wound healing. Oxid Med Cell Longev. 2020:49102802020. View Article : Google Scholar : PubMed/NCBI
79	Wang Y, Kuramitsu Y, Baron B, Kitagawa T, Tokuda K, Akada J, Maehara SI, Maehara Y and Nakamura K: PI3K inhibitor LY294002, as opposed to wortmannin, enhances AKT phosphorylation in gemcitabine-resistant pancreatic cancer cells. Int J Oncol. 50:606–612. 2017. View Article : Google Scholar : PubMed/NCBI
80	Lin J, Chen Y, Wei L, Hong Z, Sferra TJ and Peng J: Ursolic acid inhibits colorectal cancer angiogenesis through suppression of multiple signaling pathways. Int J Oncol. 43:1666–1674. 2013. View Article : Google Scholar : PubMed/NCBI
81	Zhang Y, Huang L, Shi H, Chen H, Tao J, Shen R and Wang T: Ursolic acid enhances the therapeutic effects of oxaliplatin in colorectal cancer by inhibition of drug resistance. Cancer Sci. 109:94–102. 2018. View Article : Google Scholar : PubMed/NCBI
82	Zeng Q, Che Y, Zhang Y, Chen M, Guo Q and Zhang W: Thymol isolated from thymus vulgaris L. inhibits colorectal cancer cell growth and metastasis by suppressing the Wnt/β-catenin pathway. Drug Des Devel Ther. 14:2535–2547. 2020. View Article : Google Scholar : PubMed/NCBI
83	Yao M, Ma X, Zhang X, Shi L, Liu T, Liang X, Zhao H, Li X, Li L, Gao H, et al: Lectin-mediated pH-sensitive doxorubicin prodrug for pre-targeted chemotherapy of colorectal cancer with enhanced efficacy and reduced side effects. Theranostics. 9:747–760. 2019. View Article : Google Scholar : PubMed/NCBI
84	O'Bryan RM, Baker LH, Gottlieb JE, Rivkin SE, Balcerzak SP, Grumet GN, Salmon SE, Moon TE and Hoogstraten B: Dose response evaluation of adriamycin in human neoplasia. Cancer. 39:1940–1948. 1977. View Article : Google Scholar : PubMed/NCBI
85	Gabizon A, Shmeeda H and Barenholz Y: Pharmacokinetics of pegylated liposomal doxorubicin: Review of animal and human studies. Clin Pharmacokinet. 42:419–436. 2003. View Article : Google Scholar : PubMed/NCBI
86	Marina NM, Cochrane D, Harney E, Zomorodi K, Blaney S, Winick N, Bernstein M and Link MP: Dose escalation and pharmacokinetics of pegylated liposomal doxorubicin (Doxil) in children with solid tumors: A pediatric oncology group study. Clin Cancer Res. 8:413–418. 2002.PubMed/NCBI
87	Nair AB and Jacob S: A simple practice guide for dose conversion between animals and human. J Basic Clin Pharm. 7:27–31. 2016. View Article : Google Scholar : PubMed/NCBI
88	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2019. CA Cancer J Clin. 69:7–34. 2019. View Article : Google Scholar : PubMed/NCBI
89	Xie YH, Chen YX and Fang JY: Comprehensive review of targeted therapy for colorectal cancer. Signal Transduct Target Ther. 5:222020. View Article : Google Scholar : PubMed/NCBI
90	Li J, Ma X, Chakravarti D, Shalapour S and DePinho RA: Genetic and biological hallmarks of colorectal cancer. Genes Dev. 35:787–820. 2021. View Article : Google Scholar : PubMed/NCBI
91	Tiwari A, Saraf S, Verma A, Panda PK and Jain SK: Novel targeting approaches and signaling pathways of colorectal cancer: An insight. World J Gastroenterol. 24:4428–4435. 2018. View Article : Google Scholar : PubMed/NCBI
92	Krishnamurthy N and Kurzrock R: Targeting the Wnt/beta-catenin pathway in cancer: Update on effectors and inhibitors. Cancer Treat Rev. 62:50–60. 2018. View Article : Google Scholar : PubMed/NCBI
93	Chen Z, Oh D, Dubey AK, Yao M, Yang B, Groves JT and Sheetz M: EGFR family and Src family kinase interactions: Mechanics matters? Curr Opin Cell Biol. 51:97–102. 2018. View Article : Google Scholar : PubMed/NCBI
94	Lopez A, Harada K, Vasilakopoulou M, Shanbhag N and Ajani JA: Targeting angiogenesis in colorectal carcinoma. Drugs. 79:63–74. 2019. View Article : Google Scholar : PubMed/NCBI
95	Ahmad R, Singh JK, Wunnava A, Al-Obeed O, Abdulla M and Srivastava SK: Emerging trends in colorectal cancer: Dysregulated signaling pathways (Review). Int J Mol Med. 47:142021. View Article : Google Scholar : PubMed/NCBI
96	Kassi E, Sourlingas TG, Spiliotaki M, Papoutsi Z, Pratsinis H, Aligiannis N and Moutsatsou P: Ursolic acid triggers apoptosis and Bcl-2 downregulation in MCF-7 breast cancer cells. Cancer Invest. 27:723–733. 2009. View Article : Google Scholar : PubMed/NCBI
97	Yim EK, Lee KH, Namkoong SE, Um SJ and Park JS: Proteomic analysis of ursolic acid-induced apoptosis in cervical carcinoma cells. Cancer Lett. 235:209–220. 2006. View Article : Google Scholar : PubMed/NCBI
98	Argenziano M, Gigliotti CL, Clemente N, Boggio E, Ferrara B, Trotta F, Pizzimenti S, Barrera G, Boldorini R, Bessone F, et al: Improvement in the anti-tumor efficacy of doxorubicin nanosponges in in vitro and in mice bearing breast tumor models. Cancers (Basel). 12:1622020. View Article : Google Scholar : PubMed/NCBI
99	Brattain MG, Brattain DE, Fine WD, Khaled FM, Marks ME, Kimball PM, Arcolano LA and Danbury BH: Initiation and characterization of cultures of human colonic carcinoma with different biological characteristics utilizing feeder layers of confluent fibroblasts. Oncodev Biol Med. 2:355–366. 1981.PubMed/NCBI
100	Brattain MG, Fine WD, Khaled FM, Thompson J and Brattain DE: Heterogeneity of malignant cells from a human colonic carcinoma. Cancer Res. 41:1751–1756. 1981.PubMed/NCBI
101	Ahmed D, Eide PW, Eilertsen IA, Danielsen SA, Eknaes M, Hektoen M, Lind GE and Lothe RA: Epigenetic and genetic features of 24 colon cancer cell lines. Oncogenesis. 2:e712013. View Article : Google Scholar : PubMed/NCBI
102	Bazan V, Migliavacca M, Zanna I, Tubiolo C, Grassi N, Latteri MA, La Farina M, Albanese I, Dardanoni G, Salerno S, et al: Specific codon 13 K-ras mutations are predictive of clinical outcome in colorectal cancer patients, whereas codon 12 K-ras mutations are associated with mucinous histotype. Ann Oncol. 13:1438–1446. 2002. View Article : Google Scholar : PubMed/NCBI
103	Lupertz R, Watjen W, Kahl R and Chovolou Y: Dose- and time-dependent effects of doxorubicin on cytotoxicity, cell cycle and apoptotic cell death in human colon cancer cells. Toxicology. 271:115–121. 2010. View Article : Google Scholar : PubMed/NCBI
104	Nie W, Zan X, Yu T, Ran M, Hong Z, He Y, Yang T, Ju Y and Gao X: Synergetic therapy of glioma mediated by a dual delivery system loading α-mangostin and doxorubicin through cell cycle arrest and apoptotic pathways. Cell Death Dis. 11:9282020. View Article : Google Scholar : PubMed/NCBI
105	Tilija Pun N, Jang WJ and Jeong CH: Role of autophagy in regulation of cancer cell death/apoptosis during anti-cancer therapy: Focus on autophagy flux blockade. Arch Pharm Res. 43:475–488. 2020. View Article : Google Scholar : PubMed/NCBI
106	Lin YJ, Liang WM, Chen CJ, Tsang H, Chiou JS, Liu X, Cheng CF, Lin TH, Liao CC, Huang SM, et al: Network analysis and mechanisms of action of Chinese herb-related natural compounds in lung cancer cells. Phytomedicine. 58:1528932019. View Article : Google Scholar : PubMed/NCBI
107	Doğan Şiğva ZÖ, Balci Okcanoğlu T, Biray Avci Ç, Yilmaz Süslüer S, Kayabaşi Ç, Turna B, Dodurga Y, Nazli O and Gündüz C: Investigation of the synergistic effects of paclitaxel and herbal substances and endemic plant extracts on cell cycle and apoptosis signal pathways in prostate cancer cell lines. Gene. 687:261–271. 2019. View Article : Google Scholar : PubMed/NCBI
108	Aiello P, Sharghi M, Mansourkhani SM, Ardekan AP, Jouybari L, Daraei N, Peiro K, Mohamadian S, Rezaei M, Heidari M, et al: Medicinal plants in the prevention and treatment of colon cancer. Oxid Med Cell Longev. 2019:20756142019. View Article : Google Scholar : PubMed/NCBI
109	Phan T, Nguyen VH, A'Lincourt Salazar M, Wong P, Diamond DJ, Yim JH and Melstrom LG: Inhibition of autophagy amplifies baicalein-induced apoptosis in human colorectal cancer. Mol Ther Oncolytics. 19:1–7. 2020. View Article : Google Scholar : PubMed/NCBI
110	Mandal S, Gamit N, Varier L, Dharmarajan A and Warrier S: Inhibition of breast cancer stem-like cells by a triterpenoid, ursolic acid, via activation of Wnt antagonist, sFRP4 and suppression of miRNA-499a-5p. Life Sci. 265:1188542021. View Article : Google Scholar : PubMed/NCBI
111	Zheng JL, Wang SS, Shen KP, Chen L, Peng X, Chen JF, An HM and Hu B: Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells. BMC Complement Med Ther. 21:522021. View Article : Google Scholar : PubMed/NCBI
112	Yang S, Zhang X, Qu H, Qu B, Yin X and Zhao H: Cabozantinib induces PUMA-dependent apoptosis in colon cancer cells via AKT/GSK-3β/NF-κB signaling pathway. Cancer Gene Ther. 27:368–377. 2020. View Article : Google Scholar : PubMed/NCBI
113	Qin J, Fu M, Wang J, Huang F, Liu H, Huangfu M, Yu D, Liu H, Li X, Guan X and Chen X: PTEN/AKT/mTOR signaling mediates anticancer effects of epigallocatechin-3-gallate in ovarian cancer. Oncol Rep. 43:1885–1896. 2020.PubMed/NCBI
114	Zhu ML, Zhang PM, Jiang M, Yu SW and Wang L: Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells. BMC Complement Med Ther. 20:2092020. View Article : Google Scholar : PubMed/NCBI
115	Li W, Li C, Ma L and Jin F: Resveratrol inhibits viability and induces apoptosis in the small-cell lung cancer H446 cell line via the PI3K/Akt/c-Myc pathway. Oncol Rep. 44:1821–1830. 2020.PubMed/NCBI
116	Tian J, Zhang H, Mu L, Wang M, Li X, Zhang X, Xie E, Ma M, Wu D and Du Y: The miR-218/GAB2 axis regulates proliferation, invasion and EMT via the PI3K/AKT/GSK-3β pathway in prostate cancer. Exp Cell Res. 394:1121282020. View Article : Google Scholar : PubMed/NCBI
117	Qi X, Sun L, Wan J, Xu R, He S and Zhu X: Tensin4 promotes invasion and migration of gastric cancer cells via regulating AKT/GSK-3β/snail signaling pathway. Pathol Res Pract. 216:1530012020. View Article : Google Scholar : PubMed/NCBI
118	Chang YX, Lin YF, Chen CL, Huang MS, Hsiao M and Liang PH: Chaperonin-containing TCP-1 promotes cancer chemoresistance and metastasis through the AKT-GSK3β-β-catenin and XIAP-survivin pathways. Cancers (Basel). 12:38652020. View Article : Google Scholar : PubMed/NCBI
119	Ding L, Cao J, Lin W, Chen H, Xiong X, Ao H, Yu M, Lin J and Cui Q: The roles of cyclin-dependent kinases in cell-cycle progression and therapeutic strategies in human breast cancer. Int J Mol Sci. 21:19602020. View Article : Google Scholar : PubMed/NCBI
120	Xu S, Zhang H, Liu T, Yang W, Lv W, He D, Guo P and Li L: 6-Gingerol induces cell-cycle G1-phase arrest through AKT-GSK 3β-cyclin D1 pathway in renal-cell carcinoma. Cancer Chemother Pharmacol. 85:379–390. 2020. View Article : Google Scholar : PubMed/NCBI
121	Zhou C, Du J, Zhao L, Liu W, Zhao T, Liang H, Fang P, Zhang K and Zeng H: GLI1 reduces drug sensitivity by regulating cell cycle through PI3K/AKT/GSK3/CDK pathway in acute myeloid leukemia. Cell Death Dis. 12:2312021. View Article : Google Scholar : PubMed/NCBI
122	Zhang S, Liu X, Bawa-Khalfe T, Lu LS, Lyu YL, Liu LF and Yeh ET: Identification of the molecular basis of doxorubicin-induced cardiotoxicity. Nat Med. 18:1639–1642. 2012. View Article : Google Scholar : PubMed/NCBI
123	Desai VG, Lee T, Moland CL, Vijay V, Han T, Lewis SM, Herman EH and Fuscoe JC: Candidate early predictive plasma protein markers of doxorubicin-induced chronic cardiotoxicity in B6C3F₁ mice. Toxicol Appl Pharmacol. 363:164–173. 2019. View Article : Google Scholar : PubMed/NCBI
124	Kuenzi BM and Ideker T: Author correction: A census of pathway maps in cancer systems biology. Nat Rev Cancer. 21:2122021. View Article : Google Scholar : PubMed/NCBI
125	Kuenzi BM and Ideker T: A census of pathway maps in cancer systems biology. Nat Rev Cancer. 20:233–246. 2020. View Article : Google Scholar : PubMed/NCBI
126	Chang YC, Wu JW, Wang CW and Jang AC: Hippo signaling-mediated mechanotransduction in cell movement and cancer metastasis. Front Mol Biosci. 6:1572020. View Article : Google Scholar : PubMed/NCBI
127	Kennedy MB: Origin of PDZ (DHR, GLGF) domains. Trends Biochem Sci. 20:3501995. View Article : Google Scholar : PubMed/NCBI
128	Zheng Y and Pan D: The Hippo signaling pathway in development and disease. Dev Cell. 50:264–282. 2019. View Article : Google Scholar : PubMed/NCBI
129	Furth N and Aylon Y: The LATS1 and LATS2 tumor suppressors: Beyond the Hippo pathway. Cell Death Differ. 24:1488–1501. 2017. View Article : Google Scholar : PubMed/NCBI
130	Kim SH, Jin H, Meng RY, Kim DY, Liu YC, Chai OH, Park BH and Kim SM: Activating Hippo pathway via Rassf1 by ursolic acid suppresses the tumorigenesis of gastric cancer. Int J Mol Sci. 20:47092019. View Article : Google Scholar : PubMed/NCBI
131	Jeong SH, Kim HB, Kim MC, Lee JM, Lee JH, Kim JH, Kim JW, Park WY, Kim SY, Kim JB, et al: Hippo-mediated suppression of IRS2/AKT signaling prevents hepatic steatosis and liver cancer. J Clin Invest. 128:1010–1025. 2018. View Article : Google Scholar : PubMed/NCBI
132	Zhang S, Chen Q, Liu Q, Li Y, Sun X, Hong L, Ji S, Liu C, Geng J, Zhang W, et al: Hippo signaling suppresses cell ploidy and tumorigenesis through Skp2. Cancer Cell. 31:669–684.e7. 2017. View Article : Google Scholar : PubMed/NCBI
133	Ahmed AA, Abedalthagafi M, Anwar AE and Bui MM: Akt and Hippo pathways in Ewing's sarcoma tumors and their prognostic significance. J Cancer. 6:1005–1010. 2015. View Article : Google Scholar : PubMed/NCBI
134	Berthold R, Isfort I, Erkut C, Heinst L, Grunewald I, Wardelmann E, Kindler T, Åman P, Grünewald TGP, Cidre-Aranaz F, et al: Fusion protein-driven IGF-IR/PI3K/AKT signals deregulate Hippo pathway promoting oncogenic cooperation of YAP1 and FUS-DDIT3 in myxoid liposarcoma. Oncogenesis. 11:202022. View Article : Google Scholar : PubMed/NCBI
135	Ma W, Han C, Zhang J, Song K, Chen W, Kwon H and Wu T: The histone methyltransferase G9a promotes cholangiocarcinogenesis through regulation of the Hippo pathway kinase LATS2 and YAP signaling pathway. Hepatology. 72:1283–1297. 2020. View Article : Google Scholar : PubMed/NCBI
136	Xu W, Yang Z, Xie C, Zhu Y, Shu X, Zhang Z, Li N, Chai N, Zhang S, Wu K, et al: PTEN lipid phosphatase inactivation links the hippo and PI3K/Akt pathways to induce gastric tumorigenesis. J Exp Clin Cancer Res. 37:1982018. View Article : Google Scholar : PubMed/NCBI
137	Jang SW, Yang SJ, Srinivasan S and Ye K: Akt phosphorylates MstI and prevents its proteolytic activation, blocking FOXO3 phosphorylation and nuclear translocation. J Biol Chem. 282:30836–30844. 2007. View Article : Google Scholar : PubMed/NCBI
138	Romano D, Matallanas D, Weitsman G, Preisinger C, Ng T and Kolch W: Proapoptotic kinase MST2 coordinates signaling crosstalk between RASSF1A, Raf-1, and Akt. Cancer Res. 70:1195–1203. 2010. View Article : Google Scholar : PubMed/NCBI
139	Kim D, Shu S, Coppola MD, Kaneko S, Yuan ZQ and Cheng JQ: Regulation of proapoptotic mammalian ste20-like kinase MST2 by the IGF1-Akt pathway. PLoS One. 5:e96162010. View Article : Google Scholar : PubMed/NCBI
140	Kim SM, Ye S, Rah SY, Park BH, Wang H, Kim JR, Kim SH, Jang KY and Lee KB: RhBMP-2 activates Hippo signaling through RASSF1 in esophageal cancer cells. Sci Rep. 6:268212016. View Article : Google Scholar : PubMed/NCBI
141	Pankova D, Jiang Y, Chatzifrangkeskou M, Vendrell I, Buzzelli J, Ryan A, Brown C and O'Neill E: RASSF1A controls tissue stiffness and cancer stem-like cells in lung adenocarcinoma. EMBO J. 38:e1005322019. View Article : Google Scholar : PubMed/NCBI
142	Gupta V, Agarwal P and Deshpande P: Impact of RASSF1A gene methylation on clinico-pathological features of tumor and non-tumor tissue of breast cancer. Ann Diagn Pathol. 52:1517222021. View Article : Google Scholar : PubMed/NCBI
143	Lee NH, Kim SJ and Hyun J: MicroRNAs regulating Hippo-YAP signaling in liver cancer. Biomedicines. 9:3472021. View Article : Google Scholar : PubMed/NCBI
144	Agarwal S, Amin KS, Jagadeesh S, Baishay G, Rao PG, Barua NC, Bhattacharya S and Banerjee PP: Mahanine restores RASSF1A expression by down-regulating DNMT1 and DNMT3B in prostate cancer cells. Mol Cancer. 12:992013. View Article : Google Scholar : PubMed/NCBI
145	Blanchard TG, Lapidus R, Banerjee V, Bafford AC, Czinn SJ, Ahmed H and Banerjee A: Upregulation of RASSF1A in colon cancer by suppression of angiogenesis signaling and Akt activation. Cell Physiol Biochem. 48:1259–1273. 2018. View Article : Google Scholar : PubMed/NCBI