miR-519d在胰腺癌细胞中的表达及其作用

Expression and action of miR-519d in pancreatic cancer cells

目的:探讨miR-519d在胰腺癌细胞中的表达及作用。方法:用qRT-PCR检测miR-519d在胰腺癌细胞系AsPC-1、BxPC-3、Capan-2、Panc-1及正常胰腺导管上皮细胞系HPDE6-C7中的表达。将Panc-1细胞分别转染miR-519d过表达质粒(miR-519d组)与阴性对照质粒(阴性对照组),以无处理的Panc-1细胞为空白对照组,用MTT法、Transwell实验、Western blot分别检测转染后细胞的增殖和侵袭能力、凋亡情况以及X连锁凋亡抑制蛋白(XIAP)的表达。结果:各胰腺癌细胞系中miR-519d相对表达量均明显低于正常胰腺导管上皮细胞系HPDE6-C7(均P<0.05);与空白对照组比较,miR-519d组细胞增殖能力降低(培养72 h后明显降低)、凋亡率明显升高、侵袭能力明显减弱、XIAP蛋白表达量明显降低(均P<0.05);阴性对照组各项指标与空白对照组差异均无统计学意义(均P>0.05)。结论:miR-519d在胰腺癌细胞中表达下调,miR-519d表达下调有增强胰腺癌细胞增殖和侵袭能力、降低凋亡的作用,该作用可能与其调节XIAP蛋白的表达有关。

Objective： To investigate the miR-519d expression in pancreatic cancer cells, and its actions. Methods： The miR-519d expressions in pancreatic cancer cell lines AsPC-1, BxPC-3, Capan-2 and Panc-1 as well as in normal pancreatic duct epithelial cell line HPDE6-C7 were detected by qRT-PCR. The Panc-1 cells were transfected with miR-519d over-expression plasmid （miR-519d group） and negative plasmid （negative control group） respectively, with untreated Panc-1 cells as blank control group. After that, the proliferative and invasion abilities, apoptosis and expression of X-linked inhibitor of apoptosis protein （XIAP） were determined by MTT assay, Transwell assay, flow cytometry and Western blot analysis, respectively. Results： The relative expression level of miR-519d in each pancreatic cancer cell line was significantly lower than that in normal pancreatic duct epithelial HPDE6-C7 cells （all P〈0.05）. In cells of miR-519d group compared with blank control group, the proliferative ability was reduced （significantly reduced after 72-h culture）, the apoptosis rate was significantly increased, the invasion ability was significantly decreased and the expression level of XIAP protein was significantly down-regulated （all P〈0.05）. All the differences of the observed parameters in negative control group and blank control group showed no statistical significance （all P〉0.05）. Conclusion： The miR-519d expression is down-regulated in pancreatic cancer cells. Down-regulation of miR- 519d expression can enhance the proliferative and invasion abilities, and decrease apoptosis in pancreatic cancer cell lines, and these actions may be associated with its regulating XIAP protein expression.