靶向抑制 miR -19a 提高人宫颈癌 HeLa 细胞对顺铂敏感性的机制研究

摘要

Objective:To investigate the effect of miR - 19a antisense oligonucleotide(miR - 19a - ASO)on cis-platin chemosensitivity in human HeLa cells and its underlying mechanism. Methods:miR - 19a - ASO was transfect-ed to HeLa cells using Lipofectamine 2000. Real - time PCR was used to evaluate the miR - 19a expression. Cell via-bility was measured by methylthiazolyltetrazolium( MTT) assay. The expression of PTEN,AKT,and p - AKT (Ser473)protein was detected by Western blot. Results:miR - 19a - ASO inhibited the expression of miR - 19a ef-fectively. HeLa cells transfected with miR - 19a - ASO was sensitized to cisplatin(P < 0. 05)by MTT assay. Targe-ting to miR - 19a could up - regulate PTEN,and down - regulate p - AKT(Ser473)expression. Conclusion:Silence of miR - 19a increase cell sensitivity to cisplatin probably by suppressing AKT signaling pathway,providing a new tar-get for cervical cancer therapy.%目的：研究 miR -19a 反义寡核苷酸（antisense oligonucleotide，ASO）对人宫颈癌 HeLa 细胞对顺铂耐药性的影响，并进一步探讨其机制。方法：利用 Lipofectamine 2000将 miR -19a - ASO 瞬时转染 HeLa 细胞，实时定量 PCR 法检测 miR -19a 表达水平。MTT 法检测顺铂对 HeLa 细胞存活率的影响。Western blot 检测PTEN、AKT 和 p - AKT（Ser473）蛋白表达的变化。结果：miR -19a - ASO 明显抑制 HeLa 细胞 miR -19a 的表达。MTT 结果表明靶向抑制 miR -19a 能明显促进 HeLa 细胞对顺铂的敏感性。Western blot 结果提示，与对照组比较，干扰 miR -19a 的 HeLa 细胞，PTEN 表达水平升高，p - AKT（Ser473）表达水平降低。结论：沉默miR -19a 可能通过抑制 AKT 通路提高宫颈癌细胞对顺铂的敏感性，从而为宫颈癌的治疗提供新靶点。