磷脂酶Cε1在肝细胞癌中的表达及预后相关性的研究

发布时间：2018-05-09 09:40

本文选题：肝细胞癌 + 磷脂酶Cε1　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的:探究磷脂酶Cε1(Phospholipase C epsilon 1,PLCE1)在肝细胞癌组织中的表达及其对预后的预测价值。材料和方法:(1)收集90例行肝切除术且病理证实为肝细胞癌的组织石蜡切片标本及临床、预后资料,采用免疫组织化学法检测90例肝癌组织中PLCE1表达情况,统计分析PLCE1在肝细胞癌组织中的表达情况与肝细胞癌患者临床病理资料特征以及预后的相关性;(2)收集20对术后病理证实为肝细胞癌患者的新鲜癌组织和癌旁组织标本,采用定量反转录聚合酶连锁反应实验(quantificational real-time polymerase chain reaction,qRT-PCR)及蛋白免疫印记实验(western blot,WB)检测肝细胞癌组织中PLCE1 mRNA和蛋白质的表达情况;(3)采用western blot技术检测PLCE1在4种肝细胞癌细胞株(SK-Hep1、PLC、HepG2以及Hep3B)和正常肝细胞株(THLE3)的表达情况。结果:(1)免疫组织化学实验结果示在90例肝细胞癌标本中,36例(40%)癌组织中PLCE1表达下调,而癌旁组织中仅17例(18.9%)(P0.05),临床病理资料分析显示肝癌组织中PLCE1表达下调与肿瘤包膜,血管侵犯,Edmondson分级,AFP以及TNM分期密切相关(P=0.025;P=0.006;P=0.009;P=0.011;P=0.009)。生存分析中单因素分析示PLCE1表达与肝细胞癌病人根治术后的总生存率和无瘤生存率密切相关(P0.001;P0.001),多因素分析示PLCE1是肝细胞癌术后总生存率和无瘤生存率的独立危险因素(P0.001;P=0.003)。(2)qRT-PCR结果示20对新鲜组织标本中,17例(85%)标本癌组织中PLCE1的表达较癌旁组织明显下调,PLCE1在癌组织和癌旁组织中的mRNA表达量分别为0.51±0.49和1.46±1.10,差异具有统计学意义(P=0.001)。(3)WB实验结果显示,在20对新鲜组织标本中,18例(90%)标本癌组织中PLCE1的表达较癌旁组织明显下调,PLCE1在癌组织和癌旁组织中的蛋白表达量为0.93±0.39和1.22±0.40,差异具有统计学意义(P=0.026)。另外,通过WB实验显示,PLCE1蛋白在4种肝细胞癌细胞株(SK-Hep1、PLC、HepG2以及Hep3B)的表达明显低于正常肝细胞株(THLE3)。结论:PLCE1在肝细胞癌中的表达明显下调,与预后息息相关,可作为肝细胞癌患者预后的独立预测指标。
[Abstract]:Objective: to investigate the expression of phospholipase C 蔚 1(Phospholipase C epsilon 1 (PLCE1) and its prognostic value in hepatocellular carcinoma (HCC). Materials and methods paraffin sections and clinical data were collected from 90 cases of hepatectomy and confirmed by pathology. The expression of PLCE1 was detected by immunohistochemical method in 90 cases of HCC. A statistical analysis of the expression of PLCE1 in hepatocellular carcinoma (HCC) and the correlation between the clinicopathological features and prognosis of HCC patients were made. Twenty pairs of fresh and paracancerous tissues proved by postoperative pathology were collected. Quantificational real-time polymerase chain reactionation qRT-PCR and Western blotWB) were used to detect the expression of PLCE1 mRNA and protein in hepatocellular carcinoma tissues using quantitative reverse transcriptase polymerase chain reaction (QRT). Western blot technique was used to detect the expression of PLCE1 mRNA and protein in four kinds of hepatocellular carcinoma (HCC). The expression of SK-Hep1 PLCG _ 2 and Hep3B) and THLE3) in normal hepatocytes. Results the immunohistochemical results showed that the expression of PLCE1 was down-regulated in 90 hepatocellular carcinoma tissues (36 / 40), but only in 17 adjacent tissues. The clinicopathological data showed that the expression of PLCE1 was down-regulated and the tumor capsule was found in HCC tissues. Edmondson grade of vascular invasion and TNM stage were closely related to P0. 025, P0. 006, P0. 009, P0. 011 and P0. 009. In survival analysis, univariate analysis showed that the expression of PLCE1 was closely related to the overall survival rate and tumor-free survival rate of patients with hepatocellular carcinoma after radical operation. Multivariate analysis showed that PLCE1 was an independent risk of overall survival and tumor-free survival after operation for hepatocellular carcinoma. The results of RT-PCR showed that the expression of PLCE1 in cancer tissues of 20 pairs of fresh tissues was significantly lower than that of adjacent tissues (0.51 卤0.49 and 1.46 卤1.10, respectively), and the difference was statistically significant. The expression of PLCE1 in cancer tissues was significantly lower than that in adjacent tissues (0.93 卤0.39 and 1.22 卤0.40, respectively), and the difference was statistically significant. In addition, the expression of PLCE1 protein in four kinds of hepatocellular carcinoma cell lines (SK-Hep1, HepG2 and Hep3B) was significantly lower than that in normal hepatocytes. Conclusion the expression of 1% PLCE1 is down-regulated in hepatocellular carcinoma, which is closely related to the prognosis. It can be used as an independent predictor of the prognosis of hepatocellular carcinoma.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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