Shikonin potentiates paclitaxel antitumor efficacy in esophageal cancer cells via the apoptotic pathway

Du,Wenzhen; Hao,Xiaohong; Yuan,Zhili; Wang,Ying; Zhang,Xueguang; Liu,Jie

doi:10.3892/ol.2019.10662

Shikonin potentiates paclitaxel antitumor efficacy in esophageal cancer cells via the apoptotic pathway

Authors:
- Wenzhen Du
- Xiaohong Hao
- Zhili Yuan
- Ying Wang
- Xueguang Zhang
- Jie Liu
View Affiliations

Affiliations: Department of Gastroenterology, Yantai Yeda Hospital, Yantai, Shandong 264000, P.R. China, Department of Hematology and Oncology, Yantai Yeda Hospital, Yantai, Shandong 264000, P.R. China, Department of Otolaryngology, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China, Department of Neurosurgery, Liaocheng People's Hospital, Liaocheng, Shandong 252000, P.R. China
Published online on: July 25, 2019 https://doi.org/10.3892/ol.2019.10662
Pages: 3195-3201
Copyright: © Du 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

Shikonin is a natural naphthoquinone pigment that can suppress the growth of a number of cancer cell types. Paclitaxel is an antineoplastic chemotherapy drug, which is used for the treatment of various types of solid tumor cancer. However, acquired paclitaxel resistance results in the failure of therapy, and consequent metastasis and relapse. The aim of the present study was to investigate whether shikonin can sensitize esophageal cancer cells to paclitaxel‑treatment and to elucidate the underlying mechanisms. The biological effects of these two agents on esophageal cancer cell lines KYSE270 and KYSE150 were investigated by MTT assay, cell cycle analysis, Annexin‑V apoptosis assay, western blotting and reverse transcription‑quantitative polymerase chain reaction. The results demonstrated that shikonin could significantly increase the cell growth inhibition effect induced by paclitaxel in the examined cell lines (P<0.001). The addition of shikonin to paclitaxel promoted cancer cell mitotic arrest and induced significantly higher levels of cell apoptosis. Notably, the mRNA and protein levels of Bcl‑2 were downregulated, while p53 was upregulated in KYSE270 and KYSE150 cells following combined treatment. In summary, shikonin can sensitize esophageal cancer cells to paclitaxel‑treatment by promoting cell mitotic arrest and reinforcing the susceptibility of esophageal cancer cells to apoptosis induced by paclitaxel, which is potentially associated with altered levels of Bcl‑2 and p53.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2019. CA Cancer J Clin. 69:7–34. 2019. View Article : Google Scholar : PubMed/NCBI
2	Zhang Y: Epidemiology of esophageal cancer. World J Gastroenterol. 19:5598–5606. 2013. View Article : Google Scholar : PubMed/NCBI
3	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
4	Law S and Wong J: Changing disease burden and management issues for esophageal cancer in the Asia-Pacific region. J Gastroenterol Hepatol. 17:374–381. 2002. View Article : Google Scholar : PubMed/NCBI
5	Holmes RS and Vaughan TL: Epidemiology and pathogenesis of esophageal cancer. Semin Radiat Oncol. 17:2–9. 2007. View Article : Google Scholar : PubMed/NCBI
6	Stanton RA, Gernert KM, Nettles JH and Aneja R: Drugs that target dynamic microtubules: A new molecular perspective. Med Res Rev. 31:443–481. 2011. View Article : Google Scholar : PubMed/NCBI
7	Yvon AM, Wadsworth P and Jordan MA: Taxol suppresses dynamics of individual microtubules in living human tumor cells. Mol Biol Cell. 10:947–959. 1999. View Article : Google Scholar : PubMed/NCBI
8	Blagosklonny MV: Prolonged mitosis versus tetraploid checkpoint: How p53 measures the duration of mitosis. Cell Cycle. 5:971–975. 2006. View Article : Google Scholar : PubMed/NCBI
9	Dumontet C and Jordan MA: Microtubule-binding agents: A dynamic field of cancer therapeutics. Nat Rev Drug Discov. 9:790–803. 2010. View Article : Google Scholar : PubMed/NCBI
10	Ikui AE, Yang CP, Matsumoto T and Horwitz SB: Low concentrations of taxol cause mitotic delay followed by premature dissociation of p55CDC from Mad2 and BubR1 and abrogation of the spindle checkpoint, leading to aneuploidy. Cell Cycle. 4:1385–1388. 2005. View Article : Google Scholar : PubMed/NCBI
11	Orr GA, Verdier-Pinard P, McDaid H and Horwitz SB: Mechanisms of Taxol resistance related to microtubules. Oncogene. 22:7280–7295. 2003. View Article : Google Scholar : PubMed/NCBI
12	Hou Y, Guo T, Wu C, He X and Zhao M: Effect of shikonin on human breast cancer cells proliferation and apoptosis in vitro. Yakugaku Zasshi. 126:1383–1386. 2006. View Article : Google Scholar : PubMed/NCBI
13	Han W, Li L, Qiu S, Lu Q, Pan Q, Gu Y, Luo J and Hu X: Shikonin circumvents cancer drug resistance by induction of a necroptotic death. Mol Cancer Ther. 6:1641–1649. 2007. View Article : Google Scholar : PubMed/NCBI
14	Yang H, Zhou P, Huang H, Chen D, Ma N, Cui QC, Shen S, Dong W, Zhang X, Lian W, et al: Shikonin exerts antitumor activity via proteasome inhibition and cell death induction in vitro and in vivo. Int J Cancer. 124:2450–2459. 2009. View Article : Google Scholar : PubMed/NCBI
15	Chang IC, Huang YJ, Chiang TI, Yeh CW and Hsu LS: Shikonin induces apoptosis through reactive oxygen species/extracellular signal-regulated kinase pathway in osteosarcoma cells. Biol Pharm Bull. 33:816–824. 2010. View Article : Google Scholar : PubMed/NCBI
16	Long S, GuangZhi Y, BaoJie G, Wei X, YanYong H, YingLi W, Yang Z and LiHua L: Shikonin derivatives protect immune organs from damage and promote immune responses in vivo in tumour-bearing mice. Phytother Res. 26:26–33. 2012. View Article : Google Scholar : PubMed/NCBI
17	Zhao Q, Assimopoulou AN, Klauck SM, Damianakos H, Chinou I, Kretschmer N, Rios JL, Papageorgiou VP, Bauer R and Efferth T: Inhibition of c-MYC with involvement of ERK/JNK/MAPK and AKT pathways as a novel mechanism for shikonin and its derivatives in killing leukemia cells. Oncotarget. 6:38934–38951. 2015. View Article : Google Scholar : PubMed/NCBI
18	Liu X and Sun G: Shikonin enhances Adriamycin antitumor effects by inhibiting efflux pumps in A549 cells. Oncol Lett. 14:4270–4276. 2017. View Article : Google Scholar : PubMed/NCBI
19	Song J, Zhao Z, Fan X, Chen M, Cheng X, Zhang D, Wu F, Ying X and Ji J: Shikonin potentiates the effect of arsenic trioxide against human hepatocellular carcinoma in vitro and in vivo. Oncotarget. 7:70504–70515. 2016. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Mukhtar E, Adhami VM and Mukhtar H: Targeting microtubules by natural agents for cancer therapy. Mol Cancer Ther. 13:275–284. 2014. View Article : Google Scholar : PubMed/NCBI
22	Elmore S: Apoptosis: A review of programmed cell death. Toxicol Pathol. 35:495–516. 2007. View Article : Google Scholar : PubMed/NCBI
23	Henley D, Isbill M, Fernando R, Foster JS and Wimalasena J: Paclitaxel induced apoptosis in breast cancer cells requires cell cycle transit but not Cdc2 activity. Cancer Chemother Pharmacol. 59:235–249. 2007. View Article : Google Scholar : PubMed/NCBI
24	Januchowski R, Zawierucha P, Andrzejewska M, Ruciński M and Zabel M: Microarray-based detection and expression analysis of ABC and SLC transporters in drug-resistant ovarian cancer cell lines. Biomed Pharmacother. 67:240–245. 2013. View Article : Google Scholar : PubMed/NCBI
25	Sun QL, Sha HF, Yang XH, Bao GL, Lu J and Xie YY: Comparative proteomic analysis of paclitaxel sensitive A549 lung adenocarcinoma cell line and its resistant counterpart A549-Taxol. J Cancer Res Clin Oncol. 137:521–532. 2011. View Article : Google Scholar : PubMed/NCBI
26	Zhao Q, Kretschmer N, Bauer R and Efferth T: Shikonin and its derivatives inhibit the epidermal growth factor receptor signaling and synergistically kill glioblastoma cells in combination with erlotinib. Int J Cancer. 137:1446–1456. 2015. View Article : Google Scholar : PubMed/NCBI
27	Li W, Liu J, Jackson K, Shi R and Zhao Y: Sensitizing the therapeutic efficacy of taxol with shikonin in human breast cancer cells. PLoS One. 9:e940792014. View Article : Google Scholar : PubMed/NCBI
28	Wang Z, Yin J, Li M, Shen J, Xiao Z, Zhao Y, Huang C, Zhang H, Zhang Z, Cho CH and Wu X: Combination of shikonin with paclitaxel overcomes multidrug resistance in human ovarian carcinoma cells in a P-gp-independent manner through enhanced ROS generation. Chin Med. 14:72019. View Article : Google Scholar : PubMed/NCBI
29	Yang Q, Li S, Fu Z, Lin B, Zhou Z, Wang Z, Hua Y and Cai Z: Shikonin promotes adriamycininduced apoptosis by upregulating caspase-3 and caspase-8 in osteosarcoma. Mol Med Rep. 16:1347–1352. 2017. View Article : Google Scholar : PubMed/NCBI
30	Danial NN and Korsmeyer SJ: Cell death: Critical control points. Cell. 116:205–219. 2004. View Article : Google Scholar : PubMed/NCBI
31	Aravind L, Dixit VM and Koonin EV: Apoptotic molecular machinery: Vastly increased complexity in vertebrates revealed by genome comparisons. Science. 291:1279–1284. 2001. View Article : Google Scholar : PubMed/NCBI
32	Ogura T, Tanaka Y, Tamaki H and Harada M: Docetaxel induces Bcl-2- and pro-apoptotic caspase-independent death of human prostate cancer DU145 cells. Int J Oncol. 48:2330–2338. 2016. View Article : Google Scholar : PubMed/NCBI
33	Rong YP, Barr P, Yee VC and Distelhorst CW: Targeting Bcl-2 based on the interaction of its BH4 domain with the inositol 1,4,5-trisphosphate receptor. Biochim Biophys Acta. 1793:971–978. 2009. View Article : Google Scholar : PubMed/NCBI
34	Srivastava RK, Sasaki CY, Hardwick JM and Longo DL: Bcl-2-mediated drug resistance: Inhibition of apoptosis by blocking nuclear factor of activated T lymphocytes (NFAT)-induced Fas ligand transcription. J Exp Med. 190:253–265. 1999. View Article : Google Scholar : PubMed/NCBI
35	Yang T, Xu F, Sheng Y, Zhang W and Chen Y: A targeted proteomics approach to the quantitative analysis of ERK/Bcl-2-mediated anti-apoptosis and multi-drug resistance in breast cancer. Anal Bioanal Chem. 408:7491–7503. 2016. View Article : Google Scholar : PubMed/NCBI
36	Samuel S, Beljanski V, Van Grevenynghe J, Richards S, Ben Yebdri F, He Z, Nichols C, Belgnaoui SM, Steel C, Goulet ML, et al: BCL-2 inhibitors sensitize therapy-resistant chronic lymphocytic leukemia cells to VSV oncolysis. Mol Ther. 21:1413–1423. 2013. View Article : Google Scholar : PubMed/NCBI
37	Smith ND, Rubenstein JN, Eggener SE and Kozlowski JM: The p53 tumor suppressor gene and nuclear protein: Basic science review and relevance in the management of bladder cancer. J Urol. 169:1219–1228. 2003. View Article : Google Scholar : PubMed/NCBI
38	Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R and Beach D: p21 is a universal inhibitor of cyclin kinases. Nature. 366:701–704. 1993. View Article : Google Scholar : PubMed/NCBI
39	Yew PR and Berk AJ: Inhibition of p53 transactivation required for transformation by adenovirus early 1B protein. Nature. 357:82–85. 1992. View Article : Google Scholar : PubMed/NCBI
40	Yonish-Rouach E, Deguin V, Zaitchouk T, Breugnot C, Mishal Z, Jenkins JR and May E: Transcriptional activation plays a role in the induction of apoptosis by transiently transfected wild-type p53. Oncogene. 11:2197–2205. 1995.PubMed/NCBI
41	Wu Y, Mehew JW, Heckman CA, Arcinas M and Boxer LM: Negative regulation of bcl-2 expression by p53 in hematopoietic cells. Oncogene. 20:240–251. 2001. View Article : Google Scholar : PubMed/NCBI
42	May P and May E: Twenty years of p53 research: structural and functional aspects of the p53 protein. Oncogene. 18:7621–7636. 1999. View Article : Google Scholar : PubMed/NCBI