siRNA沉默整合素β1基因对子宫内膜癌细胞侵袭、转移的影响

Effects of si RNA Silencing Integrin β1 Gene on the Invasion and Metastasis of Endometrial Carcinoma Cells

目的:探讨si RNA沉默整合素β1基因对子宫内膜癌细胞侵袭、转移的影响。方法:选取子宫内膜癌ECC-1细胞系(ER阳性)和KLE细胞系(ER阴性),分别转染Integrinβ1 si RNA质粒(Integrinβ1 si RNA组)、无义序列si RNA质粒(无义序列对照组)和空载质粒(空载对照组),利用实时荧光定量PCR检测各组细胞中Integrinβ1 mRNA的表达,Western blot检测各组细胞中Integrinβ1、β-catenin和C-Myc蛋白的表达,Transwell小室检测各组细胞迁移和侵袭能力,MTT法检测各组细胞的增殖情况。结果:Integrinβ1 si RNA组ECC-1细胞和KLE细胞中Integrinβ1 mRNA和蛋白相对表达量均低于无义序列对照组和空载对照组(P<0.05);Integrinβ1 si RNA组ECC-1细胞和KLE细胞中β-catenin蛋白和C-Myc蛋白相对表达量均低于无义序列对照组和空载对照组,差异均有统计学意义(P<0.05);Integrinβ1 si RNA组ECC-1细胞和KLE细胞中迁移细胞数和侵袭细胞数均低于无义序列对照组和空载对照组(P<0.05);Integrinβ1 si RNA组ECC-1细胞和KLE细胞的A值均低于无义序列对照组和空载对照组(P<0.05)。结论:特异性抑制Integrinβ1基因可抑制子宫内膜癌细胞迁移、侵袭和增殖,可能与抑制Wnt信号传导有关。

Objective： To investigate the effects of si RNA silencing integrin β1 gene on the invasion and metastasis of endometrial carcinoma cells. Methods： The ECC-1 endometrial carcinoma cell line（ER-positive） and KLE cell line（ER-negative） were selected and transfected by Integrin β1 si RNA plasmid（Integrin β1 si RNA group）, onsense sequence si RNA plasmid（nonsense sequence control group） and empty vector（no load control group）. The expressions of Integrin β1 mRNA in each group were detected by using real-time PCR, the expressions of Integrin β1, β-catenin and C-Myc proteins in each group were tested by using Western blot, cell migration and invasion of different groups were detected by using Transwell chamber, the cell proliferations of different groups were tested by using MTT assay. Results： The relative expressions of Integrin β1 mRNA and Integrin β1 protein of the ECC-1 cells and KLE cells in the Integrin β1 si RNA group were relatively lower than those of the nonsense sequence control group and no load control group（P〈0.05）.The relative expressions of β-catenin proteins and C-Myc proteins of the ECC-1 cells and KLE cells in the Integrin β1 si RNA group were lower than those of the nonsense sequence control group and no load control group（P〈0.05）. The cell migration and invasion of ECC-1cells and KLE cells in the Integrin β1 si RNA group were lower than those of the nonsense sequence control group and no load control group（P〈0.05）. The A values of ECC-1 cells and KLE cells in the Integrin β1 si RNA group were lower than those of the nonsense sequence control group and no load control group（P〈0.05）. Conclusion： Specific inhibition of Integrin β1 gene expression could reduce the migration, invasion and proliferation of endometrial cancer cells, which might be related to the inhibition of Wnt signal pathway.