Actein induces apoptosis in leukemia cells through suppressing RhoA/ROCK1 signaling pathway

Zhou,Wen-Di; Wang,Xiang; Sun,Xing-Zhen; Hu,Jian; Zhang,Rong-Rong; Hong,Ze

doi:10.3892/ijo.2017.4150

Actein induces apoptosis in leukemia cells through suppressing RhoA/ROCK1 signaling pathway

Authors:
- Wen-Di Zhou
- Xiang Wang
- Xing-Zhen Sun
- Jian Hu
- Rong-Rong Zhang
- Ze Hong
View Affiliations

Affiliations: Department of Pediatrics, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China
Published online on: October 10, 2017 https://doi.org/10.3892/ijo.2017.4150
Pages: 1831-1841

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

Actein is a tetracyclic triterpenoid compound, extracted from the rhizome of Cimicifuga foetida, exhibiting anticancer activities as previously reported. However, the effects of actein on human leukemia have not been explored before. In this study, the role of actein in regulating apoptosis induction in human leukemia cells was investigated. Actein administration significantly enhanced apoptosis, especially in human leukemia cell line of U937 and the primary human leukemia cells. The promotion was accompanied by caspase-9, caspase-3 and poly(ADP-ribose) polymerase (PARP) cleavage, and cytochrome c (Cyto-c) release. Additionally, translocation of Bax into mitochondria was increased by actein, while anti-apoptotic signals of myeloid cell leukemia-1 (Mcl-1) and B cell CLL/lymphoma 2 (Bcl-2) were decreased, accompanied by reduced phosphorylated Bcl-2-associated death promoter (Bad). Furthermore, protein kinase B (AKT) activation was downregulated by actein treatment in U937 cells. RhoA, but not caspase-3, regulated Rho kinase 1 (ROCK1) expression induced by actein. Suppression of RhoA and ROCK1 reduced ROCK1 expression, caspase-9, caspase-3 and PARP cleavage. In contrast, AKT inactivity enhanced apoptosis levels, as well as caspase signaling pathway expression. The anticancer role of actein was potentiated by inactivating AKT. In vivo, U937-bearing tumor growth was suppressed by actein, which was related to ROCK1 suppression, AKT dephosphorylation and apoptosis induction. These results indicated that actein has a suppressive role in human leukemia progression through inactivating RhoA/ROCK1 and inducing caspases.

View Figures

View References

1	Shivarov V and Bullinger L: Expression profiling of leukemia patients: Key lessons and future directions. Exp Hematol. 42:651–660. 2014. View Article : Google Scholar : PubMed/NCBI
2	Ross K, Gillespie-Twardy AL, Agha M, Raptis A, Hou JZ, Farah R, Redner RL, Im A, Duggal S, Ding F, et al: Intensive chemotherapy in patients aged 70 years or older newly diagnosed with acute myeloid leukemia. Oncol Res. 22:85–92. 2015. View Article : Google Scholar : PubMed/NCBI
3	Wang ES: Treating acute myeloid leukemia in older adults. Hematology (Am Soc Hematol Educ Program). 2014:14–20. 2014.
4	Mayer J, Arthur C, Delaunay J, Mazur G, Thomas XG, Wierzbowska A, Ravandi F, Berrak E, Jones M, Li Y, et al: Multivariate and subgroup analyses of a randomized, multinational, phase 3 trial of decitabine vs treatment choice of supportive care or cytarabine in older patients with newly diagnosed acute myeloid leukemia and poor- or intermediate-risk cytogenetics. BMC Cancer. 14:692014. View Article : Google Scholar : PubMed/NCBI
5	Kantarjian HM, Thomas XG, Dmoszynska A, Wierzbowska A, Mazur G, Mayer J, Gau JP, Chou WC, Buckstein R, Cermak J, et al: Multicenter, randomized, open-label, phase III trial of decitabine versus patient choice, with physician advice, of either supportive care or low-dose cytarabine for the treatment of older patients with newly diagnosed acute myeloid leukemia. J Clin Oncol. 30:2670–2677. 2012. View Article : Google Scholar : PubMed/NCBI
6	Dombret H, Seymour JF, Butrym A, Wierzbowska A, Selleslag D, Jang JH, Kumar R, Cavenagh J, Schuh AC, Candoni A, et al: International phase 3 study of azacitidine vs conventional care regimens in older patients with newly diagnosed AML with >30% blasts. Blood. 126:291–299. 2015. View Article : Google Scholar : PubMed/NCBI
7	Zhou T, Hasty P, Walter CA, Bishop AJ, Scott LM and Rebel VI: Myelodysplastic syndrome: An inability to appropriately respond to damaged DNA? Exp Hematol. 41:665–674. 2013. View Article : Google Scholar : PubMed/NCBI
8	Einbond LS, Shimizu M, Nuntanakorn P, Seter C, Cheng R, Jiang B, Kronenberg F, Kennelly EJ and Weinstein IB: Actein and a fraction of black cohosh potentiate antiproliferative effects of chemotherapy agents on human breast cancer cells. Planta Med. 72:1200–1206. 2006. View Article : Google Scholar : PubMed/NCBI
9	Einbond LS, Mighty J, Redenti S and Wu HA: Actein induces calcium release in human breast cancer cells. Fitoterapia. 91:28–38. 2013. View Article : Google Scholar : PubMed/NCBI
10	Yue GGL, Xie S, Lee JKM, Kwok HF, Gao S, Nian Y, Wu XX, Wong CK, Qiu MH and Lau CB: New potential beneficial effects of actein, a triterpene glycoside isolated from Cimicifuga species, in breast cancer treatment. Sci Rep. 6:352632016. View Article : Google Scholar : PubMed/NCBI
11	Yang ZC and Ma J: Actein enhances TRAIL effects on suppressing gastric cancer progression by activating p53/caspase-3 signaling. Biochem Biophys Res Commun. Nov 30–2016.Epub ahead of print. View Article : Google Scholar
12	Einbond LS, Shimizu M, Ma H, Wu HA, Goldsberry S, Sicular S, Panjikaran M, Genovese G and Cruz E: Actein inhibits the Na⁺-K⁺-ATPase and enhances the growth inhibitory effect of digitoxin on human breast cancer cells. Biochem Biophys Res Commun. 375:608–613. 2008. View Article : Google Scholar : PubMed/NCBI
13	Harris SL and Levine AJ: The p53 pathway: Positive and negative feedback loops. Oncogene. 24:2899–2908. 2005. View Article : Google Scholar : PubMed/NCBI
14	Lin MW, Wu CT, Shih JY, Chang YL and Yang PC: Clinicopathologic characteristics and prognostic significance of EGFR and p53 mutations in surgically resected lung adenocarcinomas ≤2 cm in maximal dimension. J Surg Oncol. 110:99–106. 2014. View Article : Google Scholar : PubMed/NCBI
15	Xu Y, Wang L, Zheng X, Liu G, Wang Y, Lai X and Li J: Positive expression of p53, c-erbB2 and MRP proteins is correlated with survival rates of NSCLC patients. Mol Clin Oncol. 1:487–492. 2013. View Article : Google Scholar
16	Lee JH, Katakai T, Hara T, Gonda H, Sugai M and Shimizu A: Roles of p-ERM and Rho-ROCK signaling in lymphocyte polarity and uropod formation. J Cell Biol. 167:327–337. 2004. View Article : Google Scholar : PubMed/NCBI
17	Osiak AE, Zenner G and Linder S: Subconfluent endothelial cells form podosomes downstream of cytokine and RhoGTPase signaling. Exp Cell Res. 307:342–353. 2005. View Article : Google Scholar : PubMed/NCBI
18	Benitah SA, Valerón PF and Lacal JC: ROCK and nuclear factor-kappaB-dependent activation of cyclooxygenase-2 by Rho GTPases: Effects on tumor growth and therapeutic consequences. Mol Biol Cell. 14:3041–3054. 2003. View Article : Google Scholar : PubMed/NCBI
19	Cai L, Threadgill MD, Wang Y and Li M: Effect of poly (ADP-ribose) polymerase-1 inhibition on the proliferation of murine colon carcinoma CT26 cells. Pathol Oncol Res. 15:323–328. 2009. View Article : Google Scholar
20	Ratnam K and Low JA: Current development of clinical inhibitors of poly(ADP-ribose) polymerase in oncology. Clin Cancer Res. 13:1383–1388. 2007. View Article : Google Scholar : PubMed/NCBI
21	Takeba Y, Matsumoto N, Watanabe M, Takenoshita-Nakaya S, Ohta Y, Kumai T, Takagi M, Koizumi S, Asakura T and Otsubo T: The Rho kinase inhibitor fasudil is involved in p53-mediated apoptosis in human hepatocellular carcinoma cells. Cancer Chemother Pharmacol. 69:1545–1555. 2012. View Article : Google Scholar : PubMed/NCBI
22	Tortora G, Caputo R, Damiano V, Caputo R, Troiani T, Veneziani BM, De Placido S, Bianco AR, Zangemeister-Wittke U and Ciardiello F: Combined targeted inhibition of bcl-2, bcl-XL, epidermal growth factor receptor, and protein kinase A type I causes potent antitumor, apoptotic, and antiangiogenic activity. Clin Cancer Res. 9:866–871. 2003.PubMed/NCBI
23	Riedl SJ and Shi Y: Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol. 5:897–907. 2004. View Article : Google Scholar : PubMed/NCBI
24	Yang JY, Della-Fera MA, Rayalam S and Baile CA: Enhanced effects of xanthohumol plus honokiol on apoptosis in 3T3-L1 adipocytes. Obesity (Silver Spring). 16:1232–1238. 2008. View Article : Google Scholar
25	Roy S, Yu Y, Padhye SB, Sarkar FH and Majumdar AP: Difluorinated-curcumin (CDF) restores PTEN expression in colon cancer cells by down-regulating miR-21. PLoS One. 8:e685432013. View Article : Google Scholar : PubMed/NCBI
26	Mueller S, Phillips J, Onar-Thomas A, Romero E, Zheng S, Wiencke JK, McBride SM, Cowdrey C, Prados MD, Weiss WA, et al: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome. Neurooncol. 14:1146–1152. 2012.
27	Liu Y, Minze LJ, Mumma L, Li XC, Ghobrial RM and Kloc M: Mouse macrophage polarity and ROCK1 activity depend on RhoA and non-apoptotic caspase 3. Exp Cell Res. 341:225–236. 2016. View Article : Google Scholar : PubMed/NCBI
28	Gilkes DM, Xiang L, Lee SJ, Chaturvedi P, Hubbi ME, Wirtz D and Semenza GL: Hypoxia-inducible factors mediate coordinated RhoA-ROCK1 expression and signaling in breast cancer cells. Proc Natl Acad Sci USA. 111:E384–E393. 2014. View Article : Google Scholar :
29	Wang Y, Wang D and Guo D: miR-124 promote neurogenic transdifferentiation of adipose derived mesenchymal stromal cells partly through RhoA/ROCK1, but not ROCK2 signaling pathway. PLoS One. 11:e01466462016. View Article : Google Scholar : PubMed/NCBI
30	Zhang JG, Li XY, Wang YZ, Zhang QD, Gu SY, Wu X, Zhu GH, Li Q and Liu GL: ROCK is involved in vasculogenic mimicry formation in hepatocellular carcinoma cell line. PLoS One. 9:e1076612014. View Article : Google Scholar : PubMed/NCBI
31	Peng H, Luo P, Li Y, Wang C, Liu X, Ye Z, Li C and Lou T: Simvastatin alleviates hyperpermeability of glomerular endothelial cells in early-stage diabetic nephropathy by inhibition of RhoA/ROCK1. PLoS One. 8:e800092013. View Article : Google Scholar : PubMed/NCBI
32	Lin SC, Gou GH, Hsia CW, Ho CW, Huang KL, Wu YF, Lee SY and Chen YH: Simulated microgravity disrupts cytoskeleton organization and increases apoptosis of rat neural crest stem cells via upregulating CXCR4 expression and RhoA-ROCK1-p38 MAPK-p53 signaling. Stem Cells Dev. 25:1172–1193. 2016. View Article : Google Scholar : PubMed/NCBI
33	Altieri DC: Survivin and IAP proteins in cell-death mechanisms. Biochem J. 430:199–205. 2010. View Article : Google Scholar : PubMed/NCBI
34	Mohan S, Abdul AB, Abdelwahab SI, Al-Zubairi AS, Sukari MA, Abdullah R, Elhassan Taha MM, Ibrahim MY and Syam S: Typhonium flagelliforme induces apoptosis in CEMss cells via activation of caspase-9, PARP cleavage and cytochrome c release: Its activation coupled with G0/G1 phase cell cycle arrest. J Ethnopharmacol. 131:592–600. 2010. View Article : Google Scholar : PubMed/NCBI
35	Liu X, Zou H, Slaughter C and Wang X: DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell. 89:175–184. 1997. View Article : Google Scholar : PubMed/NCBI
36	Grütter MG: caspases: Key players in programmed cell death. Curr Opin Struct Biol. 10:649–655. 2000. View Article : Google Scholar : PubMed/NCBI
37	Wang Y, Xu J and Gu Y: 37 1, 25 (OH) 37 1,25(OH)2D3 suppresses oxidative stress-induced microparticle release by placental trophoblasts: Placenta and decidua. Pregnancy Hyperens. 6:1542016. View Article : Google Scholar
38	Shen K, Wang Y, Zhang Y, Zhou H, Song Y, Cao Z, Kou J and Yu B: Cocktail of four active components derived from Sheng Mai San inhibits hydrogen peroxide-induced PC12 cell apoptosis linked with the caspase-3/ROCK1/MLC pathway. Rejuvenation Res. 18:517–527. 2015. View Article : Google Scholar : PubMed/NCBI
39	Li G, Zhou T, Liu L, Chen J, Zhao Z, Peng Y, Li P and Gao N: Ezrin dephosphorylation/downregulation contributes to ursolic acid-mediated cell death in human leukemia cells. Blood Cancer J. 3:e1082013. View Article : Google Scholar : PubMed/NCBI
40	Peng H, Cao J, Yu R, Danesh F, Wang Y, Mitch WE, Xu J and Hu Z: CKD stimulates muscle protein loss via rho-associated protein kinase 1 activation. J Am Soc Nephrol. 27:509–519. 2016. View Article : Google Scholar :
41	Heath-Engel HM, Chang NC and Shore GC: The endoplasmic reticulum in apoptosis and autophagy: Role of the BCL-2 protein family. Oncogene. 27:6419–6433. 2008. View Article : Google Scholar : PubMed/NCBI
42	Martinou JC and Youle RJ: Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell. 21:92–101. 2011. View Article : Google Scholar : PubMed/NCBI
43	Levine B, Sinha S and Kroemer G: Bcl-2 family members: Dual regulators of apoptosis and autophagy. Autophagy. 4:600–606. 2008. View Article : Google Scholar : PubMed/NCBI
44	Arnoult D, Parone P, Martinou JC, Antonsson B, Estaquier J and Ameisen JC: Mitochondrial release of apoptosis-inducing factor occurs downstream of cytochrome c release in response to several proapoptotic stimuli. J Cell Biol. 159:923–929. 2002. View Article : Google Scholar : PubMed/NCBI
45	Sheridan C, Delivani P, Cullen SP and Martin SJ: Bax- or Bak-induced mitochondrial fission can be uncoupled from cytochrome c release. Mol Cell. 31:570–585. 2008. View Article : Google Scholar : PubMed/NCBI