曲古抑菌素A对卵巢癌细胞分化、增殖的影响及其机制

Effect of TSA on differentiation and proliferation of ovarian cancer cells and its mechanism

目的观察曲古抑菌素A(TSA)对卵巢癌细胞分化、增殖及细胞周期的影响,并探讨其机制。方法培养卵巢癌上皮细胞系HO8910,将细胞分为A、B、C、D组,分别加入0、100、200、400 nmol/L TSA。分别于培养24、48、72 h观察A、C组细胞形态变化;于培养24 h后采用Western blotting法检测四组细胞中的Y染色体上的性别决定区盒2(SOX-2)、八聚体结合转录因子4(OCT-4)和叉头样转录因子A2(FOXA-2)。分别采用MTT法及Ed U染色法检测细胞增殖能力,流式细胞术检测不同周期细胞。采用Western blotting法检测各组细胞中的Cyclin D1和p21Cip1蛋白。结果 A组细胞多为类圆形或椭圆形,外表规整,成簇生长。C组细胞发生形态转变,细胞密度有所下降。与A组相比,B、C、D组细胞中SOX-2、OCT-4表达下调,FOXA-2表达升高(P均<0.05)。与A组相比,B、C、D组细胞增殖率降低,G_0/G_1期细胞比例增高,S期细胞比例减小,Cyclin D1蛋白表达下调,p21Cip1蛋白表达上调(P均<0.05)。结论 TSA可诱导卵巢癌HO8910细胞分化,抑制细胞增殖,其机制可能与调节细胞分化相关蛋白及细胞周期相关蛋白表达有关。

Objective To observe the effect of trichostatin A （ TSA） on the differentiation , proliferation and cell cycle of ovarian cancer cells and to investigate its mechanism .Methods Epithelial ovarian cancer cell line HO 8910 was cul-tured and divided into groups A , B, C and D, respectively, which were added with 0, 100, 200 and 400 nmol/L TSA. Cell morphology of groups A and C were observed at 24 h, 48 h and 72 h after culture.The expression of sex-determining region Y chromosome cassette-2 （SOX-2）, octamer binding transcription factor 4 （OCT-4） and fork-like transcription fac-tor A2 （ FOXA2） in the four groups was detected by using Western blotting 24 hours after culture .The cell proliferation was detected by MTT and EdU staining , distribution of cell cycle was detected by flow cytometry and the expression of Cyc-lin D1 and p21Cip1 protein was detected by Western blotting .Results Cells in the group A were mostly round or oval , looked neat and grew in clusters .While cell morphology in the group C changed and cell density decreased .Compared with group A, the expression of SOX-2 and OCT-4 in the groups B, C and D were decreased, FOXA-2 expression was increased （all P〈0.05）.Compared with group A, the cell proliferation rates were decreased , the proportion of cells in G0/G1 phase was increased , the proportion of cells in S phase was decreased , the expression of Cyclin D 1 was decreased and the expres-sion of p21Cip1 protein in the groups B, C and D was increased （all P〈0.05）.Conclusion TSA can induce ovarian cancer cell differentiation and inhibit the cell proliferation , which may be related to the regulation of cell differentiation-re-lated and cell cycle-related proteins .