下调lncRNA RP11-333E1.2表达抑制子宫内膜癌细胞增殖和侵袭的机制研究

Inhibitory effect of down-regulating the expression of lncRNA RP11-333E1.2 on proliferation and invasion of endometrial cancer cells and its mechanism

目的检测长链非编码RNA(lncRNA)RP11-333E1.2在子宫内膜癌中的表达,观察下调RP11-333E1.2对子宫内膜癌细胞增殖和侵袭能力的影响及其调控机制。方法qPCR检测45对子宫内膜癌及癌旁组织、5种子宫内膜癌细胞株(RL95-2、Ishikawa、HEC-1B、HEC-1A、AN3CA)和1种正常子宫内膜细胞株(ESC)中RP11-333E1.2的表达。以RP11-333E1.2表达最高的子宫内膜癌细胞株为转染对象,分别转染空载质粒和可干扰RP11-333E1.2表达的siRNA质粒,命名为空载体组和siRNA组。qPCR检测转染后RP11-333E1.2表达量;MTT法和Transwell侵袭实验分别检测低表达RP11-333E1.2对子宫内膜癌细胞增殖和侵袭能力的影响;qPCR和Western blotting检测神经前体细胞表达发育下调基因9(neural precursor cell expressed,developmentally down-regulated 9,NEDD9)的mRNA和蛋白水平的表达。结果RP11-333E1.2在子宫内膜癌和癌旁组织中的相对表达水平分别为5.73±0.74和1.46±0.55,子宫内膜癌组织中RP11-333E1.2明显高表达(P<0.01)。与人正常子宫内膜细胞相比,子宫内膜癌细胞株中RP11-333E1.2明显高表达(P<0.01),Ishikawa细胞中的表达量最多(P<0.01)。与空载体组相比,siRNA组Ishikawa细胞中RP11-333E1.2表达明显降低(P<0.01)。与空载体组相比,siRNA组下调RP11-333E1.2可明显抑制Ishikawa细胞的增殖能力(P<0.05)和侵袭能力(P<0.01)。与空载体组相比,siRNA组Ishikawa细胞中NEDD9在mRNA和蛋白水平的表达明显降低(P<0.01)。结论RP11-333E1.2在子宫内膜癌中呈高表达,下调RP11-333E1.2可抑制子宫内膜癌细胞Ishikawa的增殖能力和侵袭能力,其机制可能与NEDD9基因表达改变相关。

Objective To detect the expression of long-chain non-coding RNA(lncRNA)RP11-333E1.2 in endometrial cancer,and to explore the effect of down-regulation of RP11-333E1.2 on the proliferation and invasion abilities of endometrial cancer cells and its regulatory mechanism.Methods The qPCR was used to detect the expression of RP11-333E1.2 in 45 pairs of endometrial cancer and adjacent tissues,five endometrial cancer cell lines(RL95-2,Ishikawa,HEC-1B,HEC-1A,AN3CA)and one normal endometrial cell line(ESC).The endometrial cancer cell line with the highest expression of RP11-333E1.2 was used as the transfection object,and transfected with the empty plasmid and the siRNA plasmid,respectively.The qPCR was used to detect the expression of RP11-333E1.2 after transfection.MTT assay and Transwell invasion assay were used to detect the effects of low expression of RP11-333E1.2 on the proliferation and invasion abilities of endometrial cancer cells.The qPCR and Western blotting were used to detect the expression of neural precursor cell expressed,developmentally down regulated 9(NEDD9)at the mRNA and protein levels.Results The expression level of RP11-333E1.2 in endometrial cancer tissues was significantly higher than that in adjacent tissues(5.73±0.74 vs 1.46±0.55,P<0.01).Compared with human normal endometrial epithelial cells,RP11-333E1.2 was significantly overexpressed in endometrial cancer cell lines(P<0.01),and the highest in Ishikawa cells(P<0.01).Compared with empty vector group,the expression of RP11-333E1.2 in Ishikawa cells in siRNA group was significantly reduced(P<0.01).Compared with empty vector group,low-expressed RP11-333E1.2 significantly inhibited the proliferation(P<0.05)and invasion ability(P<0.01)of Ishikawa cells in siRNA group.Compared with empty vector group,the mRNA and protein levels of NEDD9 expression in Ishikawa cells in siRNA group was significantly reduced(P<0.01).Conclusion RP11-333E1.2 is highly expressed in endometrial cancer.Down-regulating RP11-333E1.2 can inhibit the proliferation and invasion of endometrial cancer cell Ishikawa,which may be related to the change of NEDD9 gene expression.