MYEOV对人胰腺癌细胞增殖和迁移的影响

Effect of MYEOV on Proliferation and Migration of Human Pancreatic Cancer Cells

目的探讨骨髓瘤过表达基因MYEOV在人胰腺癌细胞中的表达情况及其下调对胰腺癌SW1990细胞增殖和迁移能力的影响。方法构建慢病毒载体GV248,转染胰腺癌细胞株SW1990获得SW1990-sh1和SW1990-sh2两个实验组,以空白质粒转染作为阴性对照组,未干预细胞为空白对照组。qPCR和Western blot检测转染前后MYEOV mRNA与蛋白的表达水平;CCK-8法测定细胞增殖能力;划痕实验分析细胞迁移能力。qPCR检测TGF-β/SMAD通路中相关SMADs mRNA表达水平。结果与正常胰腺导管上皮细胞系HPDE6相比,MYEOV mRNA在6株胰腺癌细胞中表达水平明显增加(P<0.01);但仅在PANC-1和SW1990细胞系中检测到低水平MYEOV蛋白表达。实验组中MYEOV mRNA和蛋白表达均明显下调。与对照组相比,SW1990细胞的增殖和迁移能力明显下降(P<0.001)。下调MYEOV能降低TGF-β通路中SMAD1、SMAD4、SMAD5和SMAD9 mRNA表达(P<0.01),而SMAD2、SMAD3和SMAD7 mRNA仅在SW1990-sh2组中表达下调(P<0.01)。结论 MYEOV在胰腺癌细胞系中转录水平高,MYEOV可通过TGF-β/SMAD通路促进胰腺癌细胞的增殖和迁移能力。

Objective To investigate the expression of MYEOV in human pancreatic cancer cells and its effect on the proliferation and migration of pancreatic cancer SW1990 cells after knockdown.Methods The lentiviral vector GV248 was constructed and infected SW1990 cells to obtain two experimental groups(SW1990-sh1 and SW1990-sh2),while the blank plasmid was transfected as a negative control group and the untreated cells were taken as blank control group.MYEOV mRNA and protein expression were detected by qPCR and Western blot before and after transfection.The cells proliferation was determined by CCK-8 assay;the cell migration ability was analyzed by scratch test;The mRNA expression of SMADs in the TGF-β/SMAD signaling pathway was detected by qPCR.Results Compared with the pancreatic ductal epithelium cell line HPDE6,the mRNA expression of MYEOV in six pancreatic cancer cells were significantly increased(P<0.01).However,low expression of MYEOV protein was detected only in PANC-1 and SW1990 cells.The mRNA and protein expression of MYEOV in the experimental groups were significantly downregulated.The proliferation and migration of SW1990 cells were lower than the control group(P<0.001).MYEOV knockdown could reduce the mRNA expression of SMAD1,SMAD4,SMAD5 and SMAD9 in TGF-βpathway,while the expression of SMAD2,SMAD3 and SMAD7 mRNA were only down-regulated in the SW1990-sh2 group(P<0.001).Conclusion MYEOV mRNA is overexpressed in pancreatic cancer cells.MYEOV could promote the proliferation and migration of pancreatic cancer cells via TGF-β/SMAD pathway.