轴突导向通路基因多态性和表达与广西肝细胞癌遗传易感性研究

发布时间：2018-06-12 21:24

  本文选题：肝细胞癌 + 轴突导向通路　； 参考：《广西医科大学》2015年博士论文

【摘要】：目的(1)研究广西扶绥壮族人群轴突导向通路PLXNC1基因ITGβ1基因、RAC1基因多态性与广西肝细胞癌遗传易感性关系；(2)探讨PLXNC1蛋白、ITGβ1蛋白SEMA7A蛋白、RAC1蛋白在肝细胞癌中的表达情况。方法(1)利用生物信息学方法筛选PLXNC1基因ITGβ1基因、RAC1基因可能有意义的SNP位点；以广西扶绥县20个肝细胞癌高发家族(79例)和10个正常对照家族(40例)为研究对象,并将研究对象分为A、B、C三组：A组：肝癌高发家系肝细胞癌患者组,以下简称A组；B组：肝癌高发家系核心成员组(非肝癌患者),以下简称B组；C组：正常对照家系人群组,以下简称C组,运用飞行时间质谱技术(MALDI-TOF MS)检测两组中PLXNC1基因ITGβ1基因、RAC1基因功能位点的基因型及等位基因频率；(2)运用免疫组化SP法检测50例肝癌组织、50例肝癌癌旁组织、40例正常肝组织中PLXNC1、 ITGβ1、SEMA7A、RAC1蛋白表达情况。结果(1)PLXNC1基因rs2272335位点经分析PLXNC1基因rs2272335位点B组人群中携带C等位基因型的个体发生HCC的风险分别是T等位基因型个体的0.47倍(95%CI=0.1-2.2),但差异无统计学意义。C组人群中携带C等位基因型的个体发生HCC的风险分别是T等位基因型个体的4.16倍(95%CI=0.37-47.3),差异有统计学意义(P=0.0320.05)。肝癌高发家族(A+B)组中C等位基因型的分布频率明显高于正常对照(C)组,差异有统计学意义(P=0.04.05,OR=7.6,95% CI:0.99-59.50),经性别、年龄、吸烟、饮酒、HBsAg等因素校正后,肝癌高发家族(A+B)组中C等位基因型的分布频率明显高于正常对照(C)组,差异有统计学意义(P=0.030.05,OR=5.09,95% CI:0.62-41.60)。B组人群中携带TC基因型的个体发生HCC的风险分别是TT基因型个体的0.68倍(95%CI=0.13-3.52),但差异无统计学意义。C组人群中携带TC基因型的个体发生HCC的风险分别是TT基因型个体的4.3倍(95%CI=0.37-50.95),但差异无统计学意义。肝癌高发家族(A+B)组中TC基因型的分布频率明显高于正常对照(C)组,但差异有无统计学意义(P=0.10 0.05,OR=5.82,95% CI:0.72-47.21),经性别、年龄、吸烟、饮酒.HBsAg等因素校正后肝癌家族聚集性的风险仍然增高(OR=4.68,95% CI:0.54-40.17,P=0.150.05)。与TT基因型相比,TC基因型在肝癌高发家族(A+B)组和正常对照(C)组间的分布频率差异无统计学意义(P=0.150.05)；(2)ITGβ1基因rs22298141位点B组人群中携带G等位基因型的个体发生HCC的风险分别是A等位基因型个体的0.67倍(95%CI=0.31-1.45)。C组人群中携带G等位基因型的个体发生HCC的风险分别是A等位基因型个体的0.75倍(95%CI=0.33-1.7),但差异无统计学意义。等位基因A与G在肝癌高发家族(A+B)组中与正常对照(C)组中的分布频率差异无统计学意义(P0.05)。B组人群中携带AG、GG基因型的个体发生HCC的风险分别是AA基因型个体的0.91倍(95%CI=0.31-2.71)和2.2倍(95%CI=0.40-11.96)。C组人群中携带AG、GG基因型的个体发生HCC的风险分别是AA基因型个体的0.67倍(95%CI=0.22-2.1)和1.05倍(95%CI=0.17-6.6)。但差异无统计学意义。肝癌高发家族(A+B)组中AG基因型的分布频率低于正常对照组(C),但差异有无统计学意义(P=0.440.05,OR=0.72,95%CI:0.32-1.65),经性别、年龄、吸烟、饮酒.HBsAg等因素校正后患肝癌的风险降低差异仍无统计学意义(OR=0.63,95%　CI:0.26-1.54,P=0.310.05)。肝癌高发家族(A+B)组中GG基因型的分布频率与正常对照(C)组比较差异无统计学意义(P=0.34 0.05,OR=0.56,95%　CI:0.18-1.82),经性别、年龄、吸烟、饮酒、HBsAg等因素校正后患肝癌的风险差异无统计学意义(OR=0.62,95%CI：0.18-2.20,P=0.460.05)；(3)RAC1基因rs6951997位点B组人群中携带G等位基因型的个体发生HCC的风险分别是T等位基因型个体的0.73倍(95%CI=0.08-6.7)。C组人群中携带G等位基因型的个体发生HCC的风险分别是T等位基因型个体的0.49倍(95%CI=0.30-8.1),但差异无统计学意义。等位基因T与G在肝癌高发家族(A+B)组中的分布频率分别为96.84%、3.16%,在正常对照(C)组中的分布频率分别为98.75%、1.25%,两组间差异无统计学意义(P0.05)。见表16RAC1基因rS 6951997位点B组人群中携带GT基因型的个体发生HCC的风险分别是TT基因型个体的0.72(95%CI=0.08-6.9)。C组人群中携带GT基因型的个体发生HCC的风险分别是TT基因型个体的2.05倍(95%CI=0.12-34.63)。但差异无统计学意义。肝癌高发家族(A+B)组中GT基因型的分布频率明显高于正常对照(C)组,但差异有无统计学意义(P=0.380.05,OR=2.64,95%CI：0.30-23.35),经性别、年龄、吸烟、饮酒、HBsAg等因素校正后患肝癌的风险仍然增高(OR=2.02,95%CI:0.21-19.88,P=0.550.05)。与TT基因型相比,GT基因型在肝癌高发家族(A+B)组和正常对照(C)组间的分布频率差异无统计学意义(P=0.550.05)。(4)肝癌组织中,PLXNC1蛋白表达水平(3.12±1.12)显著高于肝癌癌旁组织(1.54±0.67)和正常肝组织(1.23±0.87)(P0.05),但PLXNC1蛋白表达水平在肝癌癌旁组织和正常肝组织间比较无统计学差异(P0.05)。(5)肝癌组织中,ITGβ1蛋白表达水平(3.45±1.25)显著高于肝癌癌旁组织(1.76±0.98)和正常肝组织(1.02±0.56)(P0.05),在肝癌癌旁组织中ITGβ1蛋白表达水平显著高于正常肝组织(P0.05))。(6)肝癌组织中SEMA7A蛋白表达水平(3.03±1.07)显著高于肝癌癌旁组织(1.47±0.58)和正常肝组织(1.17±0.92)(P0.05),但SEMA7A蛋白表达水平在肝癌癌旁组织和正常肝组织间比较无统计学差异(P0.05)。(7)肝癌组织中,RAC1蛋白表达水平(3.25±1.14)显著高于肝癌癌旁组织(1.68±0.87)和正常肝组织(0.92±0.67)(P0.05),在肝癌癌旁组织中,RAC 1蛋白表达水平显著高于正常肝组织(P0.05)。(8)PLXNC1蛋白ITGβ1蛋白、SEMA7A蛋白、RAC1蛋白表达水平与HCC患者性别、年龄、肿块大小、HBsAg、AFP间无明显差异(P0.05)。结论1、PLXNC1基因rs2272335位点C等位基因型可能是HCC发生的风险基因型,PLXNC1基因rs2272335位点与广西肝细胞癌遗传易感性显著相关ITGβ1基因rs22298141位点多态性与广西肝细胞癌遗传易感性未发现显著相关；RAC1基因rs6951997位点多态性与广西肝细胞癌遗传易感性未发现显著相关。2轴突导向通路SEMA7A-ITGβ1-PLXNC1及相关基因(PLXNC1、JTGβ1、RAC1和SEMA7A)蛋白表达与广西肝细胞的遗传易感性可能具有相关性。
[Abstract]:Objective (1) to study the PLXNC1 gene ITG beta 1 gene of the axon guiding pathway of the Zhuang population in Guangxi, the genetic susceptibility of RAC1 gene polymorphism to the hepatocellular carcinoma in Guangxi, and (2) to explore the expression of PLXNC1 protein, ITG beta 1 protein, SEMA7A protein, RAC1 protein in hepatocellular carcinoma. (1) screening the ITG beta 1 of PLXNC1 gene by bioinformatics method. Gene, RAC1 gene may have significant SNP loci; 20 high incidence family of hepatocellular carcinoma (79 cases) and 10 normal control family (40 cases) in Fusui County, Guangxi were studied. The subjects were divided into A, B, C three groups: group A: HCC patients with hepatocellular carcinoma, hereinafter referred to as A group; B group: the core member group of HCC high hair family Liver cancer patients), the following abbreviated B group, group C: normal control family group, the following abbreviated C group, using the time of flight mass spectrometry (MALDI-TOF MS) to detect the PLXNC1 gene ITG beta 1 gene, the genotype and allele frequency of the RAC1 gene function loci; (2) 50 hepatocellular carcinoma tissues were detected by immunohistochemical SP method, and 50 liver cancer para cancer groups were detected. The expression of PLXNC1, ITG beta 1, SEMA7A, and RAC1 protein in 40 normal liver tissues. Results (1) the rs2272335 locus of the PLXNC1 gene was analyzed by the PLXNC1 gene rs2272335 site B group to carry the HCC allele of the C allele 0.47 times respectively, but the difference was not statistically significant. The risk of C allele carrying HCC in the group was 4.16 times (95%CI=0.37-47.3) of the T allele, and the difference was statistically significant (P=0.0320.05). The distribution frequency of C allele in the high incidence family (A+B) group was significantly higher than that of the normal control group (C), the difference was statistically significant (P=0.04.05, OR=7.6,95% CI:0.99-59) .50), after correction of sex, age, smoking, drinking, and HBsAg, the distribution frequency of C allele in the high incidence family (A+B) group was significantly higher than that of the normal control group (C), and the difference was statistically significant (P=0.030.05, OR=5.09,95% CI:0.62-41.60) in the.B group, the risk of HCC with TC genotype was TT genotypes, respectively. 0.68 times (95%CI=0.13-3.52), but there was no statistically significant difference in the risk of HCC in the group with TC genotype in the group.C (95%CI=0.37-50.95), but the difference was not statistically significant. The distribution frequency of TC genotypes in the high incidence family (A+B) group was significantly higher than that of the normal control group (C), but there was no difference between the group and the normal control group (C). The study significance (P=0.10 0.05, OR=5.82,95% CI:0.72-47.21), the risk of familial aggregation of liver cancer after correction of factors such as sex, age, smoking, and drinking.HBsAg was still higher (OR=4.68,95% CI:0.54-40.17, P=0.150.05). The frequency difference between the TC genotype and the normal control (C) group was different from the TT genotype. Study significance (P=0.150.05); (2) the risk of HCC with the G allele in the group B group of the rs22298141 locus B group of the ITG beta gene was 0.67 times that of the A allele (95%CI=0.31-1.45).C group, and the risk of HCC was 0.75 times that of the allele of the A allele, respectively. The difference was not statistically significant. The distribution frequency of allele A and G in HCC high incidence family (A+B) group and normal control group (C) group had no statistically significant difference (P0.05).B group with AG, and the risk of HCC in GG genotypes was 0.91 times (95%CI=0.31-2.71) and 2.2 times (95%CI=0.40-11.96) group, respectively. The risk of HCC with AG and GG genotype was 0.67 times (95%CI=0.22-2.1) and 1.05 times (95%CI=0.17-6.6) in AA genotype individuals, but the difference was not statistically significant. The distribution frequency of AG genotypes in the high incidence family (A+B) group was lower than that of the normal control group (C), but the difference was not statistically significant (P=0.440.05, OR=0.72,95%CI:0.32-1). .65), there was no significant difference in the reduction of the risk of postoperative liver cancer by sex, age, smoking, drinking.HBsAg and other factors (OR=0.63,95% CI:0.26-1.54, P=0.310.05). The distribution frequency of GG genotypes in the high risk group of liver cancer (A+B) group was not statistically significant (P=0.34 0.05, OR=0.56,95% CI:0.18-1.82), There was no significant difference in the risk of postoperative liver cancer corrected by factors such as sex, age, smoking, drinking, and HBsAg (OR=0.62,95%CI:0.18-2.20, P=0.460.05). (3) the risk of HCC in G allelic individuals with G allele in group RAC1 rs6951997 loci B group was respectively in the 0.73 times (95%CI=0.08-6.7).C group of the T allelic individuals. The risk of HCC with the G allele was 0.49 times (95%CI=0.30-8.1), but the difference was not statistically significant. The distribution frequency of T and G in the high risk group of liver cancer (A+B) group was 96.84%, 3.16%, respectively, and the distribution frequencies in the normal control (C) group were 98.75%, 1.25%, and two, respectively. There was no statistical significance (P0.05). The risk of HCC in individuals carrying GT genotypes in group B group B at the rS 6951997 locus of the table 16RAC1 gene was 2.05 times the risk of HCC of the GT genotypes in the 0.72 (95%CI=0.08-6.9).C group of the TT genotype individuals, respectively, but the difference was not statistically significant. The distribution frequency of GT genotypes in the A+B group was significantly higher than that in the normal control group (C), but the difference was statistically significant (P=0.380.05, OR=2.64,95%CI:0.30-23.35). The risk of postoperative liver cancer corrected by factors such as sex, age, smoking, drinking, HBsAg and other factors still increased (OR=2.02,95%CI:0.21-19.88, P=0.550.05). And TT gene. There was no significant difference in the distribution frequency of GT genotype between the A+B group and the normal control (C) group (P=0.550.05). (4) the expression level of PLXNC1 protein (3.12 + 1.12) was significantly higher in the liver cancer tissues (1.54 + 0.67) and normal liver tissue (1.23 + 0.87) (P0.05), but the expression level of PLXNC1 protein was in the liver cancer. There was no statistical difference between the para cancer tissue and the normal liver tissue (P0.05). (5) the expression level of ITG beta 1 protein (3.45 + 1.25) was significantly higher than that of the para cancer tissue (1.76 + 0.98) and normal liver tissue (1.02 + 0.56) (P0.05). The expression level of ITG beta 1 protein in the adjacent tissues of liver cancer was significantly higher than that of normal liver tissue (P0.05). (6) liver cancer tissue The expression level of SEMA7A protein (3.03 + 1.07) was significantly higher than that of the adjacent tissues (1.47 + 0.58) and normal liver tissue (1.17 + 0.92) (P0.05), but the expression level of SEMA7A protein was not statistically significant (P0.05). (7) the expression level of RAC1 protein (3.25 + 1.14) was significantly higher than that of the cancer side of the liver cancer. Tissue (1.68 + 0.87) and normal liver tissue (0.92 + 0.67) (P0.05), the expression level of RAC 1 protein was significantly higher than that of normal liver tissue (P0.05). (8) PLXNC1 protein ITG beta 1 protein, SEMA7A protein, RAC1 protein expression level and HCC patient sex, age, mass size, HBsAg, AFP, no significant difference (P0.05). 1, PLXNC1 gene The 272335 loci C allele may be a risk genotype of HCC. The rs2272335 locus of PLXNC1 gene is significantly related to the genetic susceptibility of Guangxi hepatocellular carcinoma, the rs22298141 locus polymorphism of the ITG beta 1 gene is not significantly correlated with the genetic susceptibility to hepatocellular carcinoma in the RAC1 gene and the genetic polymorphism of the RAC1 gene and the heredity of the hepatocellular carcinoma in Guangxi. The expression of SEMA7A-ITG beta 1-PLXNC1 and related genes (PLXNC1, JTG beta 1, RAC1 and SEMA7A) of the.2 axon oriented pathway was not found to be related to the genetic susceptibility of Guangxi liver cells.
【学位授予单位】：广西医科大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R735.7
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本文编号：2011134