欢迎访问《解剖学报》官方网站！今天是
English
RhoA/ROCKI信号通路在紫杉醇诱导宫颈癌细胞周期阻滞中的作用
Effects of RhoA/ROCKI signal pathway on the taxol-induced cell-cycle arrest of cervical cancer cells
目的 探讨RhoA/ROCKI信号通路在紫杉醇诱导宫颈癌细胞周期阻滞中的作用及其分子机制。方法 MTT法确定能引起细胞生长显著性抑制最小浓度 (1 μmol/L) 的紫杉醇。流式细胞仪检测细胞周期和RhoA蛋白表达，Western blotting检测ROCKI、细胞周期蛋白 (cyclins) 及周期依赖性激酶 (CDKs) 的表达。结果 紫杉醇处理C33A细胞后，G2/M细胞数和RhoA/ROCKI蛋白表达显著性增加，并呈时间依赖性；而cyclin B1、A、D1和CDK1蛋白表达显著性降低。上述现象被RhoA抑制剂C3转移酶全部反转；却被ROCKI蛋白抑制剂Y-27632部分反转，但与对照相比差异仍显著。紫杉醇或RhoA/ROCKI抑制剂对CDK2的表达无显著性影响。结论 紫杉醇诱导了宫颈鳞癌C33A细胞的G2/M期阻滞，其机制与RhoA活性增强后抑制特异的cyclins和CDKs有关，而下游通路中ROCKI活性仅起到部分作用。
Objective To explore the effects of RhoA/ROCKI signal pathway in taxol-induced cell cycle arrest of cervical cancer cells. Methods The MTT method was used to determine the minimal inhibition concentration of taxol in C33A cells. Cell cycle and expression of RhoA protein were assayed by flow cytometry. Expressions of ROCKI, cyclins and cyclin-dependent kinases (CDKs)were determined by Western blotting. Results After cells were treated with 1μmol/L of taxol, the cell number of G2/M phase and the expression of RhoA/ROCKI protein increased dramatically in a time-dependent manner. However, the expressions of CDK1,cyclin B1, A and D1 proteins were decreased markedly. Moreover, these effects were totally reversed when cells pretreated with the inhibitor of RhoA (C3 transferase), and partly reversed by the treatment of ROCKI inhibitor (Y-27632), although the significance still existed as compared with the control. No marked effects were observed on the expression of CDK2 in cells treated with taxol or RhoA/ROCKI inhibitors. Conclusion Taxolinduced G2/M arrest in C33A cells was related to the decreases of specific cyclins and CDK, which is tightly regulated by the increased RhoA activity, but ROCKI may only play the role partly in this process.
宫颈癌 / RhoA/ROCKI信号通路 / 细胞周期 / 周期蛋白 / 周期蛋白依赖性激酶 / 流式细胞术 / 免疫印迹法 / 人
Cervical cancer / RhoA/ROCKI signal pathway / Cell cycle / Cyclin / Cyclin-dependent kinases / Flow cytometry / Western blotting / Human
［1］Faried LS, Faried A, Kanuma T, et al. Inhibition of the mammalian target of rapamycin (mTOR) by rapamycin increases chemosensitivity of CaSki cells to paclitaxel［J］. Eur J Cancer, 2006, 42 (7):934-947.
［2］Yim EK, Tong SY, Ho EM, et al. Anticancer effects on TACC3 by treatment of paclitaxel in HPV-18 positive cervical carcinoma cells［J］. Oncol Rep, 2009, 21 (2):549-557.
［3］Bava SV, Puliappadamba VT, Deepti A, et al. Sensitization of taxol-induced apoptosis by curcumin involves down-regulation of nuclear factor-kappaB and the serine/threonine kinase Akt and is independent of tubulin polymerization［J］. J Biol Chem, 2005, 280 (8):6301-6308.
［4］Tian RQ, Wang XH, Hou LJ, et al. MicroRNA-372 is down-regulated and targets cyclin-dependent kinase 2 (CDK2) and cyclinA1 in human cervical cancer, which may contribute to tumorigenesis［J］. J Biol Chem, 2011, 286 (29):25556-25563.
［5］Rosman DS, Phukan S, Huang CC, et al. TGFBR1*6A enhances the migration and invasion of MCF-7 breast cancer cells through RhoA activation［J］. Cancer Res, 2008, 68 (5):1319-1328.
［6］Forti FL, Armelin HA. Vasopressin triggers senescence in K-ras transformed cells via RhoA-dependent downregulation of cyclin D1［J］. Endocr Relat Cancer, 2007, 14 (4):1117-1125.
［7］Bhowmick NA, Ghiassi M, Aakre M, et al. TGF-beta-induced RhoA and p160ROCK activation is involved in the inhibition of Cdc25A with resultant cell-cycle arrest［J］. Proc Natl Acad Sci USA, 2003, 100 (26):15548-15553.
［8］Wang P, Wang WL, Zhao K, et al. Effects of gap junctional channel on the Ca 2+ -mediated migration and invasion of breast cancer cells［J］. Acta Anatomica Sinica, 2012, 43 (5):624-628. (in Chinese)
王萍, 王维莉, 赵凯, 等. 缝隙连接蛋白在钙离子介导的乳腺癌细胞转移和侵袭中的作用［J］. 解剖学报, 2012, 43 (5):647-653.
［9］Zhu PX, Liu ZL, Miu J, et al. RNAi-mediated Bip knockdown improves secretion of FVIII activity by protein splicing-based two-chain gene transduced cells［J］. Acta Anatomica Sinica, 2012, 43 (5):624-628. (in Chinese)
朱普祥, 刘泽隆, 缪静, 等. RNAi介导的Bip表达下调促进基于蛋白质剪接的双链公转基因细胞分泌FVⅢ凝血活性［J］. 解剖学报, 2012, 43 (5):624-628.
［10］Wei JY, Wang Y, Hang XM, et al. Bisphosphonates preliminary study of tumor suppressor mechanism［J］. Chinese Journal of Cell Biology, 2010, 32 (4):606-612. (in Chinese)
韦金英, 王烨, 哈雪梅, 等. 双膦酸盐抑癌机制的初步研究［J］. 中国细胞生物学学报, 2010, 32 (4):606-612.
［11］Cheng HL, Su SJ, Huang LW, et al. Arecoline induces HA22T/VGH hepatoma cells to undergo anoikis-involvement of STAT3 and RhoA activation［J］. Mol Cancer, 2010, 9:126.
［12］Wang P, Chen LL, Yan H, et al. Trichosanthin suppresses HeLa cell proliferation through inhibition of the PKC/MAPK signaling pathway［J］. Cell Biol Toxicol, 2009, 25 (5):479-488.
［13］Janardhanan R, Butler JT, Banik NL, et al. N-(4-Hydroxyphenyl) retinamide potentiated paclitaxel for cell cycle arrest and apoptosis in glioblastoma C6 and RG2 cells［J］. Brain Res, 2009, 1268:142-153.
［14］Kamai T, Tsujii T, Arai K, et al. Significant association of Rho/ROCK pathway with invasion and metastasis of bladder cancer［J］. Clin Cancer Res, 2003, 9 (7):2632-2641.
［15］Guo H, Ray RM, Johnson LR. RhoA stimulates IEC-6 cell proliferation by increasing polyamine-dependent Cdk2 activity［J］. Am J Physiol Gastrointest Liver Physiol, 2003, 285 (4):G704-713.
［16］Cook DR, Solski PA, Bultman SJ, et al. The ect2 rho Guanine nucleotide exchange factor is essential for early mouse development and normal cell cytokinesis and migration［J］. Genes Cancer, 2011, 2 (10):932-942.
［17］Zhang S, Tang Q, Xu F, et al. RhoA regulates G1-S progression of gastric cancer cells by modulation of multiple INK4 family tumor suppressors［J］. Mol Cancer Res, 2009, 7 (4):570-580.
［18］Jia G, Zhang W, Zhou J, et al. Effects of paclitaxel and gefitinib on the proliferation and cell cycle of human lung adenocarcinoma cell SPC-A1［J］. Zhongguo Fei Ai Za Zhi, 2008, 11 (3):339-344.
［19］Mollah ML, Park DK, Park HJ. Cordyceps militaris Grown on Germinated Soybean Induces G2/M Cell Cycle Arrest through Downregulation of Cyclin B1 and Cdc25c in Human Colon Cancer HT-29 Cells［J］. Evid Based Complement Alternat Med, 2012, 2012:249217.
［20］Winsel S, Sommer A, Eschenbrenner J, et al. Molecular mode of action and role of TP53 in the sensitivity to the novel epothilone sagopilone (ZK-EPO) in A549 non-small cell lung cancer cells［J］. PLoS One, 2011, 6 (4):e19273.
［21］Cho HJ, Oh YJ, Han SH, et al. Cdk1-mediated phosphorylation of receptor associated protein 80 (RAP80) serine 677 residues modulate DNA damage-induced G2/M checkpoint and cell survival［J］. J Biol Chem, 2013, 288(6):3768-3776.
［22］Mac Fhearraigh S, Mc Gee MM. Cyclin B1 interacts with the BH3-only protein Bim and mediates its phosphorylation by Cdk1 during mitosis［J］. Cell Cycle, 2011, 10 (22):3886-3896.
［23］Farràs R, Baldin V, Gallach S, et al. JunB breakdown in mid-/late G2 is required for down-regulation of cyclin A2 levels and proper mitosis［J］. Mol Cell Biol, 2008, 28 (12):4173-4187.
［24］Horiuchi K, Umetani M, Minami T, et al. Wilms’tumor 1-associating protein regulates G2/M transition through stabilization of cyclin A2 mRNA［J］. Proc Natl Acad Sci USA, 2006, 103 (46):17278-17283.
［25］Khandelwal P, Padala MK, Cox J, et al. The N-terminal domain of y-box binding protein-1 induces cell cycle arrest in g2/m phase by binding to cyclin d1［J］. Int J Cell Biol, 2009, 2009:243532.
［26］Kim YM, Geiger TR, Egan DI, et al. The HTLV-1 tax protein cooperates with phosphorylated CREB, TORC2 and p300 to activate CRE-dependent cyclin D1 transcription［J］. Oncogene, 2007, 29 (14):2142-2152.
［27］Kamiya K, Sakakibara K, Ryer EJ, et al. Phosphorylation of the cyclic AMP response element binding protein mediates transforming growth factor beta-induced downregulation of cyclin A in vascular smooth muscle cells［J］. Mol Cell Biol, 2007, 27(9):3489-3498.
国家自然科学基金资助项目;市自然科学基金资助项目;自然科学基金资助项目
/
| 〈 |
|
〉 |
