siRNA抑制S100A4表达对卵巢癌细胞CP70生物学特性的影响

Effects of S100A4 Gene Silienced by S100A4 SiRNA on Cisplatin Sensitivity, Apoptosis and Migration of Ovarian Cancer Cell Line CP70

目的:探讨人卵巢癌顺铂耐药细胞株CP70沉默S100A4基因后,CP70细胞对顺铂敏感性、凋亡及细胞迁移的影响。方法:设计并合成S100A4基因特异性的siRNA并转染入卵巢癌细胞CP70,48 h后应用RT-PCR和Western Blot检测在mRNA和蛋白水平siRNA对S100A4的影响,MTT法检测转染siRNA后卵巢癌细胞CP70对顺铂敏感性的变化。用流式细胞术检测顺铂(40μM)对转染S100A4 siRNA后对卵巢癌细胞CP70凋亡的影响,Transwell法观察siRNA抑制S100A4后对卵巢癌CP70迁移能力的影响。结果:与空白对照组、阴性对照组相比,S100A4siRNA转染组CP70细胞的S100A4基因和蛋白表达降低(P<0.01)。MTT法检测顺铂敏感性发现S100A4 siRNA转染组CP70细胞顺铂敏感性增强。在顺铂刺激下,siRNA转染组细胞凋亡率高于其他各组,差异具有统计学意义(P<0.05)。Transwell发现CP70细胞迁移能力明显下降(P<0.05)。结论:S100A4 siRNA能够明显抑制CP70细胞S100A4的表达,从而增强细胞对顺铂的敏感性,促进细胞凋亡,减弱细胞的迁移能力。S100A4有望成为逆转卵巢癌铂类耐药的治疗靶点。

Objective： To observe the effect of silencing the expression of S100A4 on the cisplatin sensitivity, apoptosis and migration in cisplatin resistance cell line CP70. Methods： The S100A4 siRNA was constructed and then transfected into CP70. After 48 h, RT-PCR and Western blot were used to detect the inhibition effect of S100A4. MTT assay was performed to evaluate the effect of S100A4 gene silence on cisplatin sensitivity of CP70. The apoptosis rates were detected by using flow cytometry. Results： Compared with blank control group and negative control group,the expressions of S100A4 mRNA and protein were inhibited significantly in CP70 cells（P〈0.01） after transfecting with S100A siRNA for 48 h.MTT assay found that down-regulation of S100A4 enhanced cisplatin sensitivity in CP70. The apoptosis rate of siRNA transfection group induced by cisplatin was higher than other groups, respectively（P〈0. 05）. Cell migration ability was reduced in siRNA transfection group（P〈0.01）. Conclusion： S100A4 siRNA can significantly enhanced the cisplatin sensitivity and inhibited migration ability of CP70.S100A4 can be the target of reversing the resistance to chemotherapy in ovarian cancer.