曲古抑菌素A下调STAT3增强卵巢癌细胞顺铂化疗敏感性的体外研究
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摘要
目的探讨组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)在体外对卵巢癌顺铂(CDDP)耐药细胞C13~*增殖的抑制效果及其机制。方法 200 nmol/L TSA和20μmol/L CDDP分别单独和联合作用于人卵巢癌细胞铂耐药细胞株C13~*及其母本细胞OV2008,MTT法检测细胞生长增殖并计算存活率,流式细胞术及Western bolt检测上述药物作用时肿瘤细胞的凋亡情况;Western bolt检测C13~*及OV2008中STAT3的表达水平,siRNA下调STAT3表达后,流式细胞术检测CDDP作用时C13~*细胞的凋亡率改变。结果 MTT结果显示经TSA、CDDP或联合处理48、72 h后,与TSA和CDDP组比较,联合组能够显著抑制C13~*细胞的增殖,差异有统计学意义;流式细胞术及Western blot检测显示TSA和CDDP联合作用时,对C13~*细胞的凋亡诱导作用显著增强;铂耐药细胞C13~*中STAT3的表达水平要显著高于其母本细胞OV2008,下调STAT3的表达水平,可显著增强CDDP诱导的C13~*细胞的凋亡。结论 TSA可通过下调铂耐药细胞C13~*中STAT3蛋白的表达水平发挥对顺铂化疗的增敏作用。
Objective Our study aimed to investigate the effect of histone deacetylase inhibitor trichostatin A(TSA)on the cisplatin(CDDP) resistance of ovarian cancer cell lines and its molecular mechanism.Methods Cisplatin-resistant ovarian cancer cell line C13~* and its parental line OV2008 were incubated with TSA(200 nmol/L) or/and CDDP(20 μmol/L),the inhibitory rate of tumor cells was determined by MTT assay.Flow cytometry and Western blotting were used to detect apoptosis of tumor cells.Western blotting was also used to detect STAT3 expression in C13~* and OV2008 cells.After down-regulation of STAT3 by transfection of STAT3 siRNA in C13~* cells,cisplatin-induced apoptosis was evaluated by flow cytometry.Results MTT assay showed that the proliferation inhibitory rates of the combination group after 48 or 72 h treatment were significantly higher than those of TSA and CDDP groups.The level of STAT3 protein was much higher in C13~* cells than in OV2008 cells.Flow cytometry and Western blotting showed that TSA combined with CDDP significantly enhanced the apoptotic rate of C13~* cells.STAT3 expression level was significantly higher in C13~* cells than in OV2008.Downregulation of STAT3 can significantly improve CDDP-induced apoptosis of C13~* cells.Conclusion Down-regulation of STAT3 by TSA endows cisplatin-resistant cells C13~* with increased sensitivity to cisplatin.
Objective Our study aimed to investigate the effect of histone deacetylase inhibitor trichostatin A(TSA)on the cisplatin(CDDP) resistance of ovarian cancer cell lines and its molecular mechanism.Methods Cisplatin-resistant ovarian cancer cell line C13~* and its parental line OV2008 were incubated with TSA(200 nmol/L) or/and CDDP(20 μmol/L),the inhibitory rate of tumor cells was determined by MTT assay.Flow cytometry and Western blotting were used to detect apoptosis of tumor cells.Western blotting was also used to detect STAT3 expression in C13~* and OV2008 cells.After down-regulation of STAT3 by transfection of STAT3 siRNA in C13~* cells,cisplatin-induced apoptosis was evaluated by flow cytometry.Results MTT assay showed that the proliferation inhibitory rates of the combination group after 48 or 72 h treatment were significantly higher than those of TSA and CDDP groups.The level of STAT3 protein was much higher in C13~* cells than in OV2008 cells.Flow cytometry and Western blotting showed that TSA combined with CDDP significantly enhanced the apoptotic rate of C13~* cells.STAT3 expression level was significantly higher in C13~* cells than in OV2008.Downregulation of STAT3 can significantly improve CDDP-induced apoptosis of C13~* cells.Conclusion Down-regulation of STAT3 by TSA endows cisplatin-resistant cells C13~* with increased sensitivity to cisplatin.
引文
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[19] 杨琰,严兆华,叶金艳,等.叶酸受体单抗Farletuzumab对卵巢癌细胞多药耐药逆转的机制[J].华中科技大学学报:医学版,2018,47(2):183-187.
[20] 邹道养,赵爱月,林建光,等.体外诱导扩增γδT细胞对卵巢癌SKOV3细胞的影响[J].华中科技大学学报:医学版,2018,47(1):67-68,90.
[2] Dmitri P,Albandri A,Keith S et al.Histone deacetylases as new therapy targets for platinum-resistant epithelial ovarian cancer[J].J Cancer Res Clin Oncol,2016,142(8):1659-1671.
[3] Yu H,Lee H,Herrmann A,et al.Revisiting STAT3 signalling in cancer:new and unexpected biological functions[J].Nat Rev Cancer,2014,14(11):736-746.
[4] Kevin B,Nicolas G,Eric A,et al.Chemoresistance and targeted therapies in ovarian and endometrial cancers[J].Oncotarget,2017,8(3):4008-4042.
[5] Mary E,Zahra F,Aiste M,et al.DNA damage repair in ovarian cancer:unlocking the heterogeneity[J].J Ovarian Res,2018,11(1):50-62
[6] Yang Q,Yang Y,Zhou N,et al.Epigenetics in ovarian cancer:premise,properties,and perspectives[J].Mol Cancer,2018,17(1):109-130.
[7] Zou J,Yin F,Wang Q,et al:Analysis of microarray-identified genes and microRNAs associated with drug resistance in ovarian cancer[J].Int J Clin Exp Pathol,2015,8(6):6847-6858.
[8] Johnstone R W.Histone-deacetylase inhibitors:novel drugs for the treatment of cancer[J].Nat Rev Drug Discov,2002,1(4):287-299.
[9] Meng J,Zhang H,Zhou C,et al.The histone deacetylase inhibitor trichostatin A induces cell cycle arrest and apoptosis in colorectal cancer cells via p53-dependent and -independent pathways[J].Oncol Rep,2012,28(1):384-388.
[10] Wang J,Lee E,Ji M,et al.HDAC inhibitors,trichostatin A and valproic acid,increase E-cadherin and vimentin expression but inhibit migration and invasion of cholangiocarcinoma cells[J].Oncol Rep,2018,40(1):346-354.
[11] Lin W,Hsu F,Kuo K,et al.Trichostatin A,a histone de-acetylase inhibitor,induces synergistic cytotoxicity with chemotherapy via suppression of Raf/MEK/ERK pathway in urothelial carcinoma[J].J Mol Med(Berl),2018,96(12):1307-1318.
[12] Ponnusamy L,Mahalingaiah P,Chang Y,et al.Reversal of epigenetic aberrations associated with the acquisition of doxorubicin resistance restores drug sensitivity in breast cancer cells[J].Eur J Pharm Sci,2018,123:56-69.
[13] Sobue S,Mizutani N,Aoyama Y,et al.Mechanism of paclitaxel resistance in a human prostate cancer cell line,PC3-PR,and its sensitization by cabazitaxel[J].Biochem Biophys Res Commun,2016,479(4):808-813.
[14] Jin X,Fang Y,Hu Y,et al.Synergistic activity of the histone deacetylase inhibitor trichostatin A and the proteasome inhibitor PS-341 against taxane-resistant ovarian cancer cell lines[J].Oncol Lett,2017,13(6):4619-4626.
[15] Wang Z,Wang C,Zuo D,et al.Attenuation of STAT3 phosphorylation promotes apoptosis and chemosensitivity in human osteosarcoma induced by raddeanin A[J].Int J Biol Sci,2019,15(3):668-679.
[16] Wang L,Zhan X,Shen X,et al.P16 promotes the growth and mobility potential of breast cancer both in vitro and in vivo:the key role of the activation of IL-6/JAK2/STAT3 signaling[J].Mol Cell Biochem,2018,446(1/2):137-148.
[17] Onodera K,Sakurada A,Notsuda H,et al.Growth inhibition of KRAS- and EGFR-mutant lung adenocarcinoma by cosuppression of STAT3 and the SRC/ARHGAP35 axis[J].Oncol Rep,2018,40(3):1761-1768.
[18] Ta N,Chakrabandhu K,Huault S,et al.The tyrosine phosphorylated pro-survival form of Fas intensifies the EGF-induced signal in colorectal cancer cells through the nuclear EGFR/STAT3-mediated pathway[J].Sci Rep,2018,8(1):12424-12439.
[19] 杨琰,严兆华,叶金艳,等.叶酸受体单抗Farletuzumab对卵巢癌细胞多药耐药逆转的机制[J].华中科技大学学报:医学版,2018,47(2):183-187.
[20] 邹道养,赵爱月,林建光,等.体外诱导扩增γδT细胞对卵巢癌SKOV3细胞的影响[J].华中科技大学学报:医学版,2018,47(1):67-68,90.