富含亮氨酸重复序列蛋白1通过上调哺乳动物雷帕霉素靶蛋白表达促进乳腺癌细胞迁移和侵袭

Leucine-rich repeat sequence protein 1 promotes breast cancer cell migration and invasion by upregulating the expression of mammalian rapamycin target proteins

目的:探究富含亮氨酸重复序列蛋白1(leucine-rich repeat protein 1,LRR1)在乳腺癌细胞中的表达及对其生物学功能的影响。方法:培养人乳腺癌MDA-MB-231、HCC70细胞株和正常乳腺上皮MCF-10A细胞;采用蛋白免疫印迹法检测LRR1表达。将人乳腺癌MDA-MB-231、HCC70细胞株分为干扰对照组、LRR1干扰组、LRR1过表达组和空载体对照组,分别转染LRR1对照、干扰、过表达、空载体序列;采用Transwell实验检测乳腺癌细胞迁移和侵袭能力;采用蛋白免疫印迹法检测细胞中哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,m-TOR)表达。结果:与正常人乳腺上皮MCF-10A细胞相比,乳腺癌MDA-MB-231和HCC70细胞中LRR1表达水平显著增加(P均<0.01)。在人乳腺癌MDA-MB-231、HCC70细胞中,与干扰对照组相比,LRR1干扰组细胞迁移和侵袭数显著降低(P均<0.01),m-TOR表达水平明显降低(P<0.01);与空载体对照组相比,过表达LRR1组细胞迁移和侵袭数显著增加(P均<0.01),m-TOR蛋白表达水平明显增加(P均<0.01)。结论:LRR1在乳腺癌细胞中呈高表达,并可促进其迁移和侵袭。

Objective:To investigate the expression of leucine-rich repeat protein 1(LRR1)in breast cancer cells and its effect on cells biological function.Methods:Human breast cancer MDA-MB-231,HCC70 cell lines and normal breast epithelial MCF-10A cells were cultured;Western blotting was used to detect the expression of LRR1.Human breast cancer MDA-MB-231 and HCC70 cell lines were divided into interference control group,LRR1 interference group,LRR1 overexpression group and empty vector control group,which were transfected with LRR1 control,interference,overexpression and empty vector sequences,respectively;Transwell assay was used to detect migration and invasion ability of breast cancer cells;Western blotting was used to detect the expression of mammalian target of rapamycin(m-TOR).Results:Compared with normal human breast epithelial MCF-10A cells,the expression level of LRR1 in breast cancer MDF-MB-231 and HCC70 cells was significantly increased(both P<0.01).In human breast cancer MDA-MB-231,HCC70 cell line,compared with the interference control group,LRR1 interference group significantly decreased the number of cell migration and invasion(all P<0.01),and the expression level of m-TOR was significantly decreased(both P<0.01);compared with the empty vector control group,LRR1 overexpression group significantly increased the numbers of migration and invasion(both P<0.01),and the expression level of m-TOR protein was significantly increased(both P<0.01).Conclusion:LRR1 is highly expressed in breast cancer cell lines and promotes their migration and invasion.