Holliday交叉识别蛋白在肝癌中高表达且与患者预后相关

发布时间：2018-01-29 04:09

本文关键词： Holliday 交叉识别蛋白 HJURP 肝细胞癌　出处：《大连医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:检测肝细胞癌组织中 HJURP(Holliday Junction Recognition Protein,Holliday交叉识别蛋白)的表达情况,探究其表达水平与预后的关系。方法:通过免疫组化、Westernblot、逆转录聚合酶链反应和实时聚合酶链反应等技术从蛋白和RNA水平上检测HJURP的表达水平,运用SPSS21.0(美国)软件进行统计学分析。定性资料使用卡方检验。定量资料的每组数据需先进行正态检验,符合正态分布的使用T检验或校正T检验,不符合正态分布的使用非参数检验。先比较cancer组和paracancer组。再将cancer组的患者分别按照性别、年龄、是否有饮酒史、是否有肝炎或肝硬化、临床分期、肝功能Child分级分为亚组。再对各个亚组的免疫组化、Westernblot、逆转录聚合酶链反应和实时聚合酶链反应进行分析。通过电话随访的到每位患者的无病生存期、总生存期并进行生存分析。结果:(1)PCR结果分析发现cancer组和paracancer组的Ct值分别是25.9652 ± 3.4866 和 31.1120± 7.3778。两组的 ΔCt 值分别是 4.069821 ±4.6855 和8.7879±7.07246。两组的ΔΔCt值分别是 8.4491±4.5554和 13.0779±7.0725。根据公式计算两组的相对定量值分别是0.8774±4.0691和0.6674±2.4538,P0.001。PCR结果进行亚组分析发现HJURP的表达和患者的性别、年龄、是否有肝炎或肝硬化病史、是否有饮酒史并无关系(P0.05)。但是根据临床分期将患者分为Ⅰ期、Ⅱ期、Ⅲ期组,分别有14人,17人,9人,三组的Ct值分别是27.5474±1.3655、26.6135 ± 1.0607 和 19.1424±1.3527,P1-2=0.5760、P1-30.001、P2-30.001。三组的 ΔCt 值分别是 7.6850±0.3179、4.0306±0.3517 和-5.3967±0.7401,P1-20.001、P1-30.001、P2-30.001。三组的ΔΔCt值分别是11.8607±0.1803、9.4389±0.1750和 6.8622±0.5081,P1-20.001、P1-30.001、P2-30.001。三组的相对定量值分别是 0.0003±0.0001、0.0024±0.0008 和0.6321 ±0.2679,P1-2=0.998、P1-30.001、P2-30.001。根据患者肝功能 Child-Pugh分级将患者分为A级组、B级组、C级组,分别有11人,22人,7人,三组的Ct 值分别是 26.6322±1.6156、26.6855± 1.0166 和 18.6203±1.3778,P1-2=0.9760、P1-3=0.001、P2-30.001。三组的ΔCt 值分别是 8.1055±0.2907、3.6986±0.5547和-6.1414±0.6449,P1-20.001、P1-30.001、P2-30.001。三组的ΔΔCt 值分别是 12.0627±0.1831、9.5259±0.266 和 6.85728±0.6159,P1-20.001、P1-30.001、P2-30.001。三组的相对定量值分别是 0.0003±0.0001、0.0020±0.0006和0.8125±0.3141,P1-2=0.989、P1-30.001、P2-30.001。(2)WESTERN-BLOT分析结果显示cancer组和paracancer组的灰度值分别是 1.2109±0.0660 和 0.7368±0.7503,P0.001。对肿瘤组患者的 WESTERN-BLOT亚组分析发现HJURP在蛋白水平上和性别、年龄、是否有肝炎或肝硬化病史、是否有饮酒史无关。和临床分期(0.2645±0.1024、1.1362±0.1084、1.6556±0.1020,P1-20.010,P2-3=0.04,P11-30.01)、Child 分级(1.3286±0.0388、1.3253±0.0567、1.5776±0.1417,P1-2=0.973,P2-3=0.031,P1-3=0.055)。(3)免疫组化结果显示cancer组的阴性、弱阳性、中阳性、强阳性分别是7.5%、22.5%、27.5%、42.5%;paracancer组的阴性、弱阳性、中阳性、强阳性分别是27.5%、27.5%、42.25%、2.75%(P0.001)。免疫组化的亚组分析发现HJURP在肿瘤组织中的表达水平和患者的性别(P=0.875)、年龄(P=0.092)、是否有饮酒史(P=0.461)、是否有肝炎或者肝硬化病史将(P=0.911)无关。但是和临床分期(P0.001)和肝功能Child分级(P=0.026)密切相关。(4)生存分析显示低表达组和高表达组的平均无进展生存期分别是16.3740±1.1140月,14.2830±0.6210月,P=0.0400。但是低表达组和高表达组的平均总生存期差异不明显。结论:(1)无论在RNA水平还是在蛋白质水平上,肿瘤组织中的HJURP都要明显高于癌旁组织。HJURP的表达水平和临床分期和Child分级相关,和性别、年龄、是否有饮酒史、是否有肝炎肝硬化史无关。(2)HJURP和患者的无病生存期相关,但和总生存期关系不确定。
[Abstract]:Objective: to detect HJURP in hepatocellular carcinoma (Holliday Junction Recognition Protein, Holliday protein expression recognition), and explore the relationship between expression level and prognosis. Methods: by immunohistochemistry, Westernblot, reverse transcriptase polymerase chain reaction and real-time polymerase chain reaction technology from the detection of the expression levels of HJURP protein and RNA levels the use of SPSS21.0 (America) software was used for statistical analysis. The qualitative data using the chi square test. Each set of data for quantitative data must first carry on the normality test, with normal distribution using T test or calibration T test, is not in accordance with the normal distribution of the non parametric test. Comparing the cancer group and paracancer group. The patients in the cancer group respectively according to gender, age, whether there is a history of drinking, whether hepatitis or cirrhosis, clinical stage, Child classification of liver function is divided into sub groups. The immune sub group Group, Westernblot, reverse transcriptase polymerase chain reaction and real-time polymerase chain reaction analysis. Through telephone follow-up to the disease-free survival of each patient, survival analysis and overall survival. Results: (1) the results of PCR analysis showed that cancer and paracancer groups Ct value respectively is 25.9652 + 3.4866 and 31.1120 + 7.3778. two group Ct values were 4.069821 + 4.6855 and 8.7879 + two 7.07246. group of delta delta Ct = 8.4491 + 4.5554 and 13.0779 + 7.0725. according to the formula of relative quantitative value of two groups were 0.8774 + 4.0691 and 0.6674 + 2.4538, P0.001.PCR results subgroup analysis showed that the expression of HJURP and the patients gender, age, whether there is a history of hepatitis or cirrhosis, if there is no relationship between drinking history (P0.05). But according to the clinical stage of the patients were divided into phase I, II, III group, there were 14 people, 17 people, 9 people, three group of Ct = 27.5474 + 1.3655,26.6135 + 1.0607 and 19.1424 + 1.3527, P1-2=0.5760, P1-30.001, P2-30.001. three group Ct values were 7.6850 + 0.3179,4.0306 + 0.3517 and -5.3967 + 0.7401, P1-20.001, P1-30.001, P2-30.001. three groups of delta delta Ct = 11.8607 + 0.1803,9.4389 + 0.1750 and 6.8622 + 0.5081, P1-20.001, P1-30.001. The relative quantitative P2-30.001. value of three groups were 0.0003 + 0.0001,0.0024 + 0.0008 and 0.6321 + 0.2679, P1-2=0.998, P1-30.001, P2-30.001. according to the Child-Pugh classification of liver function were divided into group A, B group, C group, there were 11 people, 22 people, 7 people, three group of Ct = 26.6322 + 1.6156,26.6855 + 1.0166 and 18.6203 + 1.3778, P1-2=0.9760, P1-3=0.001, P2-30.001. three group Ct values were 8.1055 + 0.2907,3.6986 + 0.5547 and -6.1414 + 0.6449, P1-20.001, P1-30.001, P2-30.001. three The delta delta Ct = 12.0627 + 0.1831,9.5259 + 0.266 and 6.85728 + 0.6159, P1-20.001, P1-30.001, P2-30.001. relative quantitative value of three groups were 0.0003 + 0.0001,0.0020 + 0.0006 and 0.8125 + 0.3141, P1-2=0.989, P1-30.001, P2-30.001. (2) WESTERN-BLOT analysis. The results showed that the gray scale of cancer group and paracancer group respectively 1.2109 + 0.0660 and 0.7368 + 0.7503, P0.001. of tumor patients of WESTERN-BLOT subgroup analysis found that HJURP at the protein level and gender, age, whether there is a history of hepatitis or cirrhosis, whether there is a history of drinking. And the clinical stage (0.2645 + 0.1024,1.1362 + 0.1084,1.6556 + 0.1020, P1-20.010, P2-3=0.04, P11-30.01, Child (grade) 1.3286 + 0.0388,1.3253 + 0.0567,1.5776 + 0.1417, P1-2=0.973, P2-3=0.031, P1-3=0.055). (3) immunohistochemistry results showed that cancer were negative, weak positive, positive and strong positive Were 7.5%, 22.5%, 27.5%, 42.5%; paracancer were negative, weak positive, positive and strong positive were 27.5%, 27.5%, 42.25%, 2.75% (P0.001). Immunohistochemical subgroup analysis showed that the expression level of HJURP in tumor tissues and patients' sex, age (P=0.875) (P=0.092) is there, drinking history (P=0.461), whether there is hepatitis or liver cirrhosis history (P=0.911). But the independent and clinical stage (P0.001) and Child classification of liver function (P=0.026) is closely related to (4). Survival analysis showed that low expression group and high expression group average progression free survival was 16.3740 + 1.1140 in August, 14.2830 + 0.6210 months, but the P=0.0400. low expression group and high expression group average overall survival was not significantly different. Conclusion: (1) both at the RNA level or at the protein level, the expression level and clinical tumor tissues are significantly higher in HJURP tissue adjacent to cancer.HJURP staging and Ch ILD grading correlation was not related to sex, age, drinking history, and history of hepatitis cirrhosis. (2) HJURP was associated with disease-free survival but not overall survival.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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