GJB2通过AMPK/mTOR信号通路促进宫颈癌细胞恶性增殖

GJB2 promotes malignant proliferation of cervical cancer cells through the AMPK/mTOR signaling pathway

目的筛选宫颈癌中具有临床意义的潜在药物靶点并研究此靶点蛋白在宫颈癌发生发展中的功能和潜在分子机制。方法收集2018年7月至2019年7月济南市人民医院妇科行宫颈癌手术的30例宫颈癌患者肿瘤组织标本及癌旁组织标本,通过临床数据库分别找到在宫颈癌中高表达的基因(tumor/normal>2)以及在宫颈癌中具有显著临床预后相关性的基因(pValue<0.05),将这两部分基因进行重合分析找到具有显著临床意义的靶基因,进一步通过热图分析找到在宫颈癌中相比正常组织表达差异最明显的前4个基因,通过高表达预后差原则筛选到目的靶基因;通过siRNA介导敲低目的基因的表达,利用细胞增殖实验和划痕迁移实验验证目的基因在宫颈癌发生发展中的重要性;最后通过分析与目的基因共表达基因参与的信号通路,探索目的基因在宫颈癌中发挥功能的潜在分子机制,进一步通过蛋白免疫印迹实验进行验证。结果共有78个基因在宫颈癌中高表达且具有显著临床预后相关性;GJB2在宫颈癌中显著高表达,且高表达的患者具有不良预后;敲低GJB2抑制宫颈癌细胞的生长增殖和迁移;敲低GJB2降低AMPK的磷酸化,升高mTOR的磷酸化,可能通过AMPK/mTOR信号通路参与宫颈癌的发生发展。结论GJB2高表达可能通过抑制AMPK的磷酸化从而升高mTOR的磷酸化参与AMPK/mTOR信号通路促进宫颈癌细胞生长增殖和迁移。

Objective To screen clinically significant potential drug targets in cervical cancer and study the function and potential molecular mechanisms of this target protein in the development of cervical cancer.Methods From July 2018 to July 2019,tumor tissue specimens and adjacent tissue specimens of 30 cases of cervical cancer patients undergoing cervical cancer surgery in the Gynecology Department of Jinan People’s Hospital were collected.According to the clinical database GEPIA,genes that are highly expressed in cervical cancer(tumor/normal>2)and genes that have significant clinical prognostic correlation in cervical cancer(pValue<0.05)were found.The overlapping analysis of these two genes was performed to find target genes with significant clinical significance,and the top 4 genes with the most significant difference in expression compared to normal tissues in cervical cancer were found through heat map analysis,and further screened by the principle of high expression and poor prognosis.Knocked down the expression of the target gene through siRNA,used cell proliferation experiments and cell migration experiments to verify the importance of the target gene in the development of cervical cancer.Finally,by analyzing the signal pathways involved in co-expressing genes with the target gene,the potential molecular mechanism of the target gene’s function in cervical cancer was explored,and further verified by Western blot experiments.Results A total of 78 genes were highly expressed in cervical cancer and have significant clinical prognosis;GJB2 was significantly highly expressed in cervical cancer,and patients with high expression had a poor prognosis;Knockdown of GJB2 inhibited the growth,proliferation and migration of cervical cancer cells;GJB2 may participate in the occurrence and development of cervical cancer through the AMPK/mTOR signaling pathway.Conclusion GJB2 overexpression inhibits AMPK phosphorylation and increases mTOR phosphorylation,thereby participating in the AMPK/mTOR signaling pathway to promote cervical cancer cell growth,proliferation and migration.