过表达CHL1基因对乳腺癌细胞瘤细胞活力、侵袭能力和凋亡的影响及其作用机制

Effects of overexpression of CHL1 gene on tumor cell viability,invasive ability and apoptosis of the breast cancer cell and its mechanism

目的:探讨过表达细胞黏附样分子(close homologue of L1,CHL1)基因对乳腺癌细胞增殖、凋亡及侵袭的影响及其可能的作用机制。方法:细胞实验分为正常组(细胞常规培养,不进行外界干预)、对照组(细胞转染pcDNA3.1-CHL1-mimic-NC后,常规培养)、实验组(细胞转染pcDNA3.1-CHL1-mimic后,常规培养);流式细胞术检测各组细胞的凋亡率,Transwell小室检测细胞的侵袭能力。Western blot检测各组细胞中CHL1、分化抗原44(CD44)表达的水平。结果:单因素方差分析显示,正常组、对照组和实验组细胞凋亡率、侵袭细胞个数及CHL1、CD44表达水平的差异均具有统计意义(F=2698.000,P=0.000;F=655.973,P=0.000;F=914.107,P=0.000;F=3653.000,P=0.000)。流式细胞结果显示实验组细胞的凋亡率明显高于对照组(P=0.000);Transwell小室显示实验组细胞的细胞侵袭个数明显少于对照组(P=0.000);Western blot检测显示实验组细胞中CHL1的表达明显高于正常组和对照组(P=0.000),CD44表达明显低于正常组和对照组(P=0.000)。结论:过表达CHL1能够抑制乳腺癌细胞增殖与侵袭能力,促使其凋亡,这可能与特异性结合CD44有关。

Objective:To investigate the effect and mechanism of overexpression of close homologue of L1(CHL1)gene on tumor cell viability,invasive ability and apoptosis of the breast cancer cell.Methods:The cell experiments were divided into normal group(cells were routinely cultured without external intervention),control group(cells were transfected with pcDNA3.1-CHL1-mimic-NC,conventional culture),and experimental group(cells were transfected with pcDNA3.1-CHL1-mimic,routine culture).Flow cytometry was used to detect the apoptosis rate of each group of cells,and Transwell cell was used to detect the cell invasion ability.Western blot was used to detect the expression level of CHL1 and cluster of differentiation antigen 44(CD44)in cells of each group.Results:One-way analysis of variance showed that the differences in the apoptosis rate,the number of invaded cells and the expression levels of CHL1 and CD44 in the normal group,control group and experimental group were statistically significant(F=2698.000,P=0.000;F=655.973,P=0.000;F=914.107,P=0.000;F=3653.000,P=0.000).The results of flow cytometry showed that the apoptosis rate of the experimental group was significantly higher than that of the control group(P=0.000);the Transwell chamber showed that the number of cell invasion of the experimental group was significantly less than that of the control group(P=0.000);Western blot detection showed the expression of CHL1 in the cells of the experimental group was significantly higher than that of the normal group and the control group(P=0.000),and the expression of CD44 was significantly lower than that of the normal group and the control group(P=0.000).Conclusion:Overexpression of CHL1 can inhibit the proliferation and invasion ability of breast cancer cells and promote their apoptosis,which may be related to the specific binding of CD44.