连接蛋白43通过调控细胞周期抑制结直肠癌的增殖机制研究

Connexin 43 inhibiting the proliferation of colorectal cancer by regulating the cell cycle

目的探究连接蛋白(CX43)在结直肠癌中的表达和临床关系及其作用与机制。方法在线分析网站检索GJA1在结直肠癌中的表达及其与结直肠癌的临床特征关系,下载TCGA数据库中的结直肠癌转录组数据,通过生物信息学方法分析GJA1在配对结直肠癌中的表达,收集临床结直肠癌样本,通过qPCR、Western blotting及免疫组织化学技术验证CX43在结直肠癌中的表达情况。在过表达和敲低CX43的HCT116和LOVO细胞中,利用克隆形成实验及CCK8实验检验其增殖能力,通过流式细胞仪检测其对细胞周期的影响,进一步裸鼠成瘤实验在体内验证其作用,最后通过qPCR和Western blotting技术验证其作用机制。结果TCGA数据库和组织样本中发现CX43在结直肠癌中低表达,且与更差的预后相关。过表达GJA1的HCT116细胞转录组数据分析显示,其可以通过抑制细胞周期来抑制结直肠癌的增殖,过表达和敲低CX43能够通过调控细胞周期蛋白D1(CCND1)的水平来抑制或促进结直肠癌的增殖。结论CX43在结直肠癌中低表达,并能够通过抑制细胞周期来抑制结直肠癌的增殖。

Objective To investigate the expression and clinical relevance of Connexin 43(CX43)in colorectal cancer,as well as its role and mechanisms in colorectal cancer.Methods The expression of GJA1 in colorectal cancer and its association with clinical characteristics of colorectal cancer were analyzed on online websites.The transcriptomic data of colorectal cancer from the TCGA database was downloaded,and bioinformatics methods were utilized to analyze the expression of GJA1 in paired colorectal cancer.Clinical colorectal cancer samples were collected for validation of CX43 expression through qPCR,Western blotting,and immunohistochemistry techniques.Transcriptional profiling was performed by overexpressing GJA1 in HCT116 cells to explore its role and mechanisms in colorectal cancer.The proliferative capacity of HCT116 and LOVO cells with overexpressed or knocked-down CX43 was assessed through colony formation and CCK8 assays.Flow cytometry was employed to examine the effects of CX43 on the cell cycle.The in vivo effects of CX43 were validated through xenograft experiments in nude mice.Finally,the mechanisms underlying the observed effects were verified using qPCR and Western blotting techniques.Results Low expression of CX43 in colorectal cancer was identified in the TCGA database and tissue samples,and the low expression of GJA1 was associated with poor prognosis in colorectal cancer.Transcriptomic analysis of HCT116 cells overexpressing GJA1 revealed its ability to inhibit colorectal cancer proliferation by D0I:10.38770793.2024.05.006 suppressing the cell cycle.Overexpression and knockdown of CX43 can inhibit or promote the proliferation of colorectal cancer through the regulation of CCND1 levels.Conclusion GJA1 is downregulated in colorectal cancer and can inhibit the proliferation of colorectal cancer by suppressing the cell cycle.