MBL2作为男性肝癌患者预后标志物的分析与验证

Analysis and validation of MBL2 as a prognostic marker of male hepatocellular carcinoma

目的探究甘露糖结合凝集素2(MBL2)基因表达对肝细胞癌(HCC)细胞系增殖和预后的影响。方法采用生物信息学方法分析MBL2在不同性别HCC患者中的表达及其对HCC患者预后的影响。利用qRT-PCR和Western blot检测基因的mRNA和蛋白表达水平。MTT实验和细胞克隆形成实验检测细胞增殖能力;采用流式细胞术检测细胞周期和细胞凋亡的变化。基因集富集分析(GSEA)筛选MBL2富集的相关信号通路。结果生物信息学分析和qRT-PCR结果显示,MBL2在男性HCC患者中的表达高于女性患者,并且与HCC患者预后相关;与对照组比较,MBL2过表达后Huh7细胞和MHCC-97H细胞的活力和克隆形成率下降,细胞周期转化受阻,细胞凋亡数增加;GSEA和Western blot结果表明,MBL2富集在细胞周期信号通路上,过表达MBL2能够抑制CDK4蛋白表达,促进P16和BAX蛋白表达。结论MBL2在HCC组织中表达下调,与男性HCC患者预后相关,并且参与细胞增殖过程。MBL2可能是男性HCC患者治疗的潜在靶点。

Objective To investigate the expression of mannose binding lectin 2(MBL2)in hepatocellular carcinoma(HCC)cell lines and its effects on the proliferation and prognosis.Methods MBL2 expression levels in patients of different sexes and survival analysis of patients were subsequently performed using bioinformatics method.The mRNA and protein expression levels were detected with the qRT-PCR and Western blot.Cell proliferation was examined by MTT assay and cell clone formation assay;Flow cytometry was used to detect the cell apoptosis and cell cycle.Gene set enrichment analysis(GSEA)was used to analyze the signal pathways of MBL2 enrichment.Results Bioinformatics analysis and qRT-PCR results showed that MBL2 was highly expressed in male HCC patients compared with female HCC patients,and was related to the prognosis of HCC patients.Compared with the control group,over-expression of MBL2 inhibited the viability and clone formation rate of Huh7 cells and MHCC-97H cells;In addition,cell cycles were arrested and cell apoptosis increased.GSEA and Western blot results showed that MBL2 was enriched in the cell cycle signal pathway.Overexpression of MBL2 inhibited the expression of CDK4 protein and promoted the expression of P16 and BAX protein.Conclusion The expression of MBL2 is down regulated in HCC,which is related to the prognosis of male patients with HCC and participates in the process of cell proliferation.MBL2 may be a potential therapeutic target for male patients with HCC.