吉非替尼通过调控hPTTG1表达抑制卵巢癌细胞增殖并诱导凋亡的机制研究

Mechanism of gefitinib inhibiting proliferation and inducing apoptosis of ovarian cancer cells by regulating of hPTTG1

目的:吉非替尼和人垂体瘤转化基因1(hPTTG1)对卵巢癌细胞增殖和凋亡的影响及作用机制。方法:以不同浓度吉非替尼干预A2780细胞,MTT法检测细胞增殖。流式细胞仪、qRT-PCR和Western blot实验检测20μmol·L-1吉非替尼对A2780细胞凋亡及细胞中hPTTG1表达的影响。敲减hPTTG1或过表达hPTTG1联合20μmol·L-1吉非替尼处理,qRT-PCR和Western blot、MTT和流式细胞术检测细胞中hPTTG1表达及细胞增殖、凋亡情况变化。结果:吉非替尼能够有效抑制A2780细胞增殖(P<0.05),呈浓度依赖性。以20μmol·L-1吉非替尼干预A2780细胞,细胞凋亡率升高(P<0.05),细胞中hPTTG1在mRNA和蛋白水平的表达显著下调(P<0.05)。将hPTTG1 siRNA转染至A2780细胞后,细胞hPTTG1表达水平降低(P<0.05),细胞增殖抑制率和凋亡率增大(P<0.05);过表达hPTTG1可减弱吉非替尼对A2780细胞增殖的抑制及凋亡的促进作用。结论:吉非替尼可抑制A2780细胞增殖并诱导凋亡,这一结果可能是通过抑制hPTTG1表达来完成。

OBJECTIVE To investigate the effect and mechanism of gefitinib and human pituitary tumor transforming gene 1(hPTTG1)on proliferation and apoptosis of ovarian cancer cells.METHODS Cell proliferation of A2780 cells after treatment with gefitinib at different concentrations was detected by MTT assay.The effects of gefitinib on apoptosis and expression of hPTTG1 in A2780 cells were detected by flow cytometry,qRT-PCR and Western blot,respectively.The expression of hPTTG1,cell proliferation and apoptosis of A2780 cells after transfection with hPTTG1 siRNA or overexpression of hPTTG1 combined with 20μmol·L-1 gefitinib were detected by qRT-PCR,Western blot,MTT and flow cytometry,respectively.RESULTS Gefitinib could effectively inhibit proliferation of A2780 cells in a concentration-dependent manner(P<0.05).Apoptotic rate increased(P<0.05).Treatment of A2780 cells with 20μmol·L-1 gefitinib increased the apoptosis rate(P<0.05),and the expression of hPTTG1 in the cells at the mRNA and protein levels was significantly downregulated(P<0.05).After hPTTG1 siRNA was transfected into A2780 cells,the expression level of hPTTG1 was decreased(P<0.05),and the inhibition rate of cell proliferation and apoptosis rate were increased(P<0.05);overexpression of hPTTG1 could attenuate the inhibitory effect of gefitinib on the proliferation and promotion of apoptosis of A2780 cells.CONCLUSION Gefitinib could inhibit proliferation and induce apoptosis of A2780 cells,which may be achieved by inhibiting the expression of hPTTG1.