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

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。

Targeted suppression of miR - 19a enhanced chemosensitivity of cervical cancer HeLa cells to cisplatin

Objective:To investigate the effect of miR-19a antisense oligonucleotide （miR-19a-ASO） on cisplatin chemosensitivity in human HeLa cells and its underlying mechanism.Methods:miR-19a-ASO was transfected to HeLa cells using Lipofectamine 2000.Real-time PCR was used to evaluate the miR-19a expression.Cell viability 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 effectively.HeLa cells transfected with miR-19a-ASO was sensitized to cisplatin （P<0.05） by MTT assay.Targeting 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 target for cervical cancer therapy.