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miR-200c通过靶向调节AHNAK抑制胰腺癌细胞的侵袭
作者:顾兴伟  周家华 
单位:东南大学附属中大医院 普外科, 江苏 南京 210009
关键词:miR-200c AHNAK 胰腺癌 侵袭 
分类号:R735.9
出版年·卷·期(页码):2018·37·第二期(234-238)
摘要:

目的:探讨miRNA-200c (miR-200c)对胰腺癌细胞AHNAK基因的表达及细胞侵袭能力的影响。方法:在胰腺癌细胞SW1990和PANC-1中转染miR-200c类似物(miR-200c mimics)及其阴性对照(Scramble),利用实时定量PCR检测miR-200c的表达;Transwell侵袭实验检测细胞侵袭;利用生物信息学方法预测miR-200c与AHNAK的3'非翻译区(3'UTR)结合位点;双荧光素酶报告实验验证结合能力;蛋白免疫印迹实验检测AHNAK的蛋白表达。结果:与阴性对照组相比,SW1990和PANC-1细胞转染miR-200c类似物后miR-200c的表达水平显著升高。过表达miR-200c后,显著抑制了胰腺癌细胞的侵袭能力。双荧光素酶报告实验结果显示,miR-200c能够与AHNAK的3'UTR结合;过表达miR-200c显著下调了AHNAK的蛋白表达。结论:miR-200c可以通过靶向调节AHNAK基因从而发挥抑制胰腺癌细胞侵袭的作用。

Objective:To investigate the effects of miR-200c overexpression on the expression of AHNAK and invasion of pancreatic cancer cell. Methods:The expression of miR-200c was detected by RT-PCR after transfecting with miR-200c analogue (miR-200c mimics) and negative control (Scramble) in pancreatic cancer cell lines SW1990 and PANC-1. Transwell assay was used to detect the change of cell invasion ability. We predicted AHNAK as its specific validated target gene by bioinformatics method and verified by double luciferase reporter system. The expression of AHNAK was detected by Western Blotting. Results:Compared with the negative control group (Scramble), the expression of miR-200c was significantly increased after transfecting with miR-200c analogs in SW1990 and PANC-1 cells. Overexpression of miR-200c significantly inhibited the invasion of pancreatic cancer cells. The double luciferase reporter system showed that miR-200c specifically binds to the 3'UTR region of AHNAK, and overexpression of miR-200c obviously inhibited the expression level of AHNAK in pancreatic cancer cells. Conclusion:miR-200c can inhibit the invasion of pancreatic cancer cells by targeting AHNAK.

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