LncRNA-MALAT1通过miR-142-3p/TEAD1分子轴调控结直肠癌细胞增殖与凋亡

LncRNA-MALAT1 regulates the proliferation and apoptosis of colorectal cancer cells through the miR-142-3p/TEAD1 molecular axis

目的探讨长链非编码RNA-肺腺癌转移相关转录子1(LncRNA-MALAT1)对结直肠癌细胞增殖与凋亡的影响及相关作用机制。方法通过Real-time PCR与Western blot实验检测结直肠癌细胞株与人正常结肠上皮细胞中LncRNA-MALAT1、miR-142-3p基因以及TEA结构域转录因子1(TEAD1)蛋白的表达;使用si-MALAT1转染HCT116细胞,在此基础上共转染miR-142-3p inhibitor,利用Real-time PCR与Western blot检测细胞中LncRNAMALAT1与miR-142-3p基因以及TEAD1、Bax、Bcl-2与Cyclin D1蛋白的表达,通过CCK-8实验检测细胞的增殖水平,通过流式细胞术检测细胞的凋亡水平,通过双荧光素酶实验检测LncRNA-MALAT1与miR-142-3p以及miR-142-3p与TEAD1的结合。结果与人正常结肠上皮细胞相比,结直肠癌细胞株中LncRNA-MALAT1基因与TEAD1蛋白呈高表达,miR-142-3p基因呈低表达(P<0.05);沉默LncRNA-MALAT1能够促进miR-142-3p基因与Bax蛋白表达,抑制LncRNA-MALAT1基因以及Bcl-2、Cyclin D1与TEAD1蛋白表达,抑制细胞增殖,并促进细胞凋亡;在此基础上沉默miR-142-3p能够对上述调控作用实现部分逆转;双荧光素酶实验结果显示,LncRNA-MALAT1与miR-142-3p以及miR-142-3p与TEAD1能够结合。结论 LncRNA-MALAT1能够结合miR-142-3p,促进miR-142-3p靶基因TEAD1表达,进而促进结直肠癌细胞增殖,抑制其细胞凋亡,促进结直肠癌的病理进程。

Objective To explore the effect and related mechanisms of LncRNA-MALAT1 on the proliferation and apoptosis of colorectal cancer cells.Methods The expressions of LncRNA-MALAT1 gene,miR-142-3p gene and TEAD1 protein in colorectal cancer cell lines and human normal colon epithelial cells were detected by Real-time PCR and Western blot experiments.Si-MALAT1 was transfected into HCT116 cells,and miR-142-3p inhibitor was co-transfected with si-MALAT1 into HCT116 cells on this basis.Real-time PCR and Western blot techniques were used to detect the expressions of LncRNA-MALAT1 gene,miR-142-3p gene,TEAD1 protein,Bax protein,Bcl-2 protein and Cyclin D1 protein in HCT116 cells.The level of cell proliferation was detected by CCK-8 assay,and the level of cell apoptosis in HCT116 cells was detected by flow cytometry.The dual luciferase experiment was used to detect the binding of LncRNA-MALAT1 and miR-142-3p,miR-142-3p and TEAD1 in HCT116 cells.Results Compared with human normal colon epithelial cells,LncRNA-MALAT1 gene and TEAD1 protein were expressed at high levels and miR-142-3p gene was expressed at a low level in colorectal cancer cell lines.Silencing of LncRNA-MALAT1 could promote the expressions of miR-142-3p gene and Bax protein,inhibit the expressions of LncRNA-MALAT1 gene,Bcl-2 protein,Cyclin D1 protein and TEAD1 protein,inhibit the level of cell proliferation and promote the level of cell apoptosis in HCT116 cells.Co-silencing of miR-142-3p and LncRNA-MALAT1 could partially reverse the above regulatory effects.The binding of LncRNA-MALAT1 and miR-142-3p,miR-142-3p and TEAD1 in HCT-116 cells were detected by the dual luciferase experiment.Conclusion LncRNA-MALAT1 could promote the expression of miR-142-3p target gene TEAD1,promote the capacity of proliferation in colorectal cancer cells,inhibit the level of apoptosis in colorectal cancer cells,and then promote the pathological process of colorectal cancer by binding to miR-142-3p.