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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 investigate the effect of cisplatin-resistant human ovarian cancer cell line CP70 on the sensitivity, apoptosis and cell migration of cisplatin (CP70) after silencing S100A4 gene. Methods: S100A4 gene-specific siRNA was designed and synthesized and transfected into ovarian cancer cell CP70 for 48 h. The mRNA and protein levels of S100A4 were detected by RT-PCR and Western Blot. The expression of S100A4 was detected by MTT assay. Change of Cancer Cell CP70 to Cisplatin Sensitivity. The effect of cisplatin (40μM) on the apoptosis of ovarian cancer cell line CP70 after transfected with S100A4 siRNA was detected by flow cytometry. The effect of S100A4 siRNA on the migration of ovarian cancer CP70 cells was observed by Transwell method. Results: Compared with the blank control group and the negative control group, S100A4 gene and protein expression in the S100A4siRNA transfected group decreased (P <0.01). The sensitivity of cisplatin to MTT assay showed that the sensitivity of cisplatin to S100A4 siRNA transfection group was enhanced. The apoptosis rate of siRNA transfection group was higher than that of other groups under the stimulation of cisplatin, the difference was statistically significant (P <0.05). Transwell found that CP70 cell migration was significantly decreased (P <0.05). Conclusion: S100A4 siRNA can significantly inhibit the expression of S100A4 in CP70 cells, thereby enhancing the sensitivity of cells to cisplatin, promoting cell apoptosis and attenuating cell migration. S100A4 is expected to become a potential therapeutic target for the reversal of platinum resistance in ovarian cancer.