BCL7A在肝细胞癌中的表达及对患者预后、肝癌细胞侵袭、迁移的影响

Expression of BCL7A in hepatocellular carcinoma and its effects on prognosis,invasion and migration of hepatocellular carcinoma

目的:分析B细胞淋巴瘤7A(BCL7A)在肝细胞癌中的表达以及与预后的关系,以及BCL7A表达对肝癌细胞侵袭、迁移的影响和机制。方法:前瞻性收集2017年11月至2018年3月于南通大学附属医院肝胆外科行根治性肝切除术的40例肝细胞癌患者的癌组织及癌旁组织用于提取蛋白质,其中男性29例,女性11例,年龄(58.5±10.4)岁。从癌症基因组图谱(TCGA)数据库中下载374例肝细胞癌和50例癌旁组织样本信息,肝癌细胞系Hep3B、SMMC-7721分别转染过表达BCL7A质粒及空载体质粒(阴性对照)。Western印迹和免疫组化检测BCL7A表达,Western印迹检测上皮间质转化相关蛋白表达(神经钙黏素、上皮钙黏素、锌指转录因子)。Transwell以及细胞划痕实验检测细胞侵袭迁移能力。结果:TCGA中50例肝细胞癌患者(癌组织与癌旁组织匹配),与癌旁组织相比,癌组织中BCL7A的mRNA相对表达显著升高,差异具有统计学意义( t=13.38, P<0.001)。依据BCL7A mRNA表达水平的中位数将374例患者分为BCL7A高表达组( n=187)和低表达组( n=187),BCL7A高表达患者术后累积生存率低于低表达组,差异有统计学意义( χ^(2)=6.95, P=0.009)。Western印迹检测肝胆外科切除的手术标本,癌组织中BCL7A蛋白相对表达也高于癌旁组织。显微镜下(×20),与对照组比较,肝癌细胞Hep3B和SMMC-7721的BCL7A过表达组侵袭的细胞数更多(153.7±1.3)个比(63.7±4.7)个、(307.7±25.1)个比(72.3±12.5)个,差异均具有统计学意义( t=7.97、8.38,均 P=0.001)。细胞划痕实验结果与Transwell侵袭检测结果一致。肝癌细胞BCL7A过表达组神经钙黏素、锌指转录因子表达高于对照组,上皮钙黏素低于对照组,差异均有统计学意义(均 P<0.05)。 结论:肝细胞癌患者癌组织中BCL7A表达升高,并且与不良预后相关。BCL7A可能通过促进上皮间质转化促进肝癌细胞转移和侵袭能力。

Objective To analyze the expression and prognosis of B-cell lymphoma 7 protein family member A(BCL7A)in hepatocellular carcinoma,as well as the effect and mechanism of BCL7A expression on the invasion and migration of hepatocellular carcinoma cells.MethodsThe cancer tissues and adjacent tissues of 40 patients with hepatocellular carcinoma who underwent radical hepatobiliary resection in the Department of Hepatobiliary Surgery,Affiliated Hospital of Nantong University from November 2017 to March 2018 were prospectively collected for protein extraction,including 29 males and 11 females,aged(58.5±10.4)years.The information of 374 cases of hepatocellular carcinoma and 50 cases of adjacent tissues were downloaded from The Cancer Genome Atlas(TCGA)database,and the hepatocellular carcinoma cell lines Hep3B and SMMC-7721 were transfected with overexpressing BCL7A plasmid and empty vector plasmid(negative control),respectively.Western blotting and immunohistochemistry were used to detect the expression of BCL7A,and Western blotting was also used to detect the expression of proteins related to epithelial-mesenchymal transition(N-cadherin,E-cadherin,snail).Transwell and cell scratch assays were used to detect cell invasion and migration.ResultsCompared with adjacent tissues,the mRNA expression of BCL7A in 50 patients with hepatocellular carcinoma in TCGA was significantly increased(t=13.38,P<0.001).According to the median mRNA expression level of BCL7A,374 patients were divided into BCL7A high expression group(n=187)and low expression group(n=187),and the cumulative survival rate of BCL7A high expression patients was lower than that of low expression group,and the difference was statistically significant(χ^(2)=6.95,P=0.009).Western blot was used to detect the relative expression of BCL7A protein in cancer tissues,and found it was higher compared to adjacent tissues.Compared with the negative control group,the number of cells invaded by the BCL7A overexpression group of hepatoma cells Hep3B and SMMC-7721 was more than the negative control group respectively,(153.7±1.3)vs(63.7±4.7)and(307.7±25.14)vs(72.3±12.5),and the differences were statistically significant(t=7.97,8.38,both P=0.001).The results of the cell scratch assay were consistent with the results of the Transwell invasion assay.The expressions of N-cadherin and snail in the BCL7A overexpression group were higher than those in the negative control group,and the E-cadherin was lower,and the difference was statistically significant(all P<0.05).ConclusionsThe expression of BCL7A in cancer tissues of patients with hepatocellular carcinoma is elevated and is associated with poor prognosis.BCL7A may promote hepatocellular carcinoma cell metastasis and invasion by promoting epithelial-mesenchymal transition.