發(fā)布時(shí)間：2018-06-12 21:24

  本文選題：肝細(xì)胞癌 + 軸突導(dǎo)向通路��； 參考：《廣西醫(yī)科大學(xué)》2015年博士論文

【摘要】：目的(1)研究廣西扶綏壯族人群軸突導(dǎo)向通路PLXNC1基因ITGβ1基因、RAC1基因多態(tài)性與廣西肝細(xì)胞癌遺傳易感性關(guān)系；(2)探討PLXNC1蛋白、ITGβ1蛋白SEMA7A蛋白、RAC1蛋白在肝細(xì)胞癌中的表達(dá)情況。方法(1)利用生物信息學(xué)方法篩選PLXNC1基因ITGβ1基因、RAC1基因可能有意義的SNP位點(diǎn)；以廣西扶綏縣20個(gè)肝細(xì)胞癌高發(fā)家族(79例)和10個(gè)正常對(duì)照家族(40例)為研究對(duì)象,并將研究對(duì)象分為A、B、C三組：A組：肝癌高發(fā)家系肝細(xì)胞癌患者組,以下簡稱A組；B組：肝癌高發(fā)家系核心成員組(非肝癌患者),以下簡稱B組；C組：正常對(duì)照家系人群組,以下簡稱C組,運(yùn)用飛行時(shí)間質(zhì)譜技術(shù)(MALDI-TOF MS)檢測兩組中PLXNC1基因ITGβ1基因、RAC1基因功能位點(diǎn)的基因型及等位基因頻率；(2)運(yùn)用免疫組化SP法檢測50例肝癌組織、50例肝癌癌旁組織、40例正常肝組織中PLXNC1、 ITGβ1、SEMA7A、RAC1蛋白表達(dá)情況。結(jié)果(1)PLXNC1基因rs2272335位點(diǎn)經(jīng)分析PLXNC1基因rs2272335位點(diǎn)B組人群中攜帶C等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是T等位基因型個(gè)體的0.47倍(95%CI=0.1-2.2),但差異無統(tǒng)計(jì)學(xué)意義。C組人群中攜帶C等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是T等位基因型個(gè)體的4.16倍(95%CI=0.37-47.3),差異有統(tǒng)計(jì)學(xué)意義(P=0.0320.05)。肝癌高發(fā)家族(A+B)組中C等位基因型的分布頻率明顯高于正常對(duì)照(C)組,差異有統(tǒng)計(jì)學(xué)意義(P=0.04.05,OR=7.6,95% CI:0.99-59.50),經(jīng)性別、年齡、吸煙、飲酒、HBsAg等因素校正后,肝癌高發(fā)家族(A+B)組中C等位基因型的分布頻率明顯高于正常對(duì)照(C)組,差異有統(tǒng)計(jì)學(xué)意義(P=0.030.05,OR=5.09,95% CI:0.62-41.60)。B組人群中攜帶TC基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是TT基因型個(gè)體的0.68倍(95%CI=0.13-3.52),但差異無統(tǒng)計(jì)學(xué)意義。C組人群中攜帶TC基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是TT基因型個(gè)體的4.3倍(95%CI=0.37-50.95),但差異無統(tǒng)計(jì)學(xué)意義。肝癌高發(fā)家族(A+B)組中TC基因型的分布頻率明顯高于正常對(duì)照(C)組,但差異有無統(tǒng)計(jì)學(xué)意義(P=0.10 0.05,OR=5.82,95% CI:0.72-47.21),經(jīng)性別、年齡、吸煙、飲酒.HBsAg等因素校正后肝癌家族聚集性的風(fēng)險(xiǎn)仍然增高(OR=4.68,95% CI:0.54-40.17,P=0.150.05)。與TT基因型相比,TC基因型在肝癌高發(fā)家族(A+B)組和正常對(duì)照(C)組間的分布頻率差異無統(tǒng)計(jì)學(xué)意義(P=0.150.05)；(2)ITGβ1基因rs22298141位點(diǎn)B組人群中攜帶G等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是A等位基因型個(gè)體的0.67倍(95%CI=0.31-1.45)。C組人群中攜帶G等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是A等位基因型個(gè)體的0.75倍(95%CI=0.33-1.7),但差異無統(tǒng)計(jì)學(xué)意義。等位基因A與G在肝癌高發(fā)家族(A+B)組中與正常對(duì)照(C)組中的分布頻率差異無統(tǒng)計(jì)學(xué)意義(P0.05)。B組人群中攜帶AG、GG基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是AA基因型個(gè)體的0.91倍(95%CI=0.31-2.71)和2.2倍(95%CI=0.40-11.96)。C組人群中攜帶AG、GG基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是AA基因型個(gè)體的0.67倍(95%CI=0.22-2.1)和1.05倍(95%CI=0.17-6.6)。但差異無統(tǒng)計(jì)學(xué)意義。肝癌高發(fā)家族(A+B)組中AG基因型的分布頻率低于正常對(duì)照組(C),但差異有無統(tǒng)計(jì)學(xué)意義(P=0.440.05,OR=0.72,95%CI:0.32-1.65),經(jīng)性別、年齡、吸煙、飲酒.HBsAg等因素校正后患肝癌的風(fēng)險(xiǎn)降低差異仍無統(tǒng)計(jì)學(xué)意義(OR=0.63,95%　CI:0.26-1.54,P=0.310.05)。肝癌高發(fā)家族(A+B)組中GG基因型的分布頻率與正常對(duì)照(C)組比較差異無統(tǒng)計(jì)學(xué)意義(P=0.34 0.05,OR=0.56,95%　CI:0.18-1.82),經(jīng)性別、年齡、吸煙、飲酒、HBsAg等因素校正后患肝癌的風(fēng)險(xiǎn)差異無統(tǒng)計(jì)學(xué)意義(OR=0.62,95%CI：0.18-2.20,P=0.460.05)；(3)RAC1基因rs6951997位點(diǎn)B組人群中攜帶G等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是T等位基因型個(gè)體的0.73倍(95%CI=0.08-6.7)。C組人群中攜帶G等位基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是T等位基因型個(gè)體的0.49倍(95%CI=0.30-8.1),但差異無統(tǒng)計(jì)學(xué)意義。等位基因T與G在肝癌高發(fā)家族(A+B)組中的分布頻率分別為96.84%、3.16%,在正常對(duì)照(C)組中的分布頻率分別為98.75%、1.25%,兩組間差異無統(tǒng)計(jì)學(xué)意義(P0.05)。見表16RAC1基因rS 6951997位點(diǎn)B組人群中攜帶GT基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是TT基因型個(gè)體的0.72(95%CI=0.08-6.9)。C組人群中攜帶GT基因型的個(gè)體發(fā)生HCC的風(fēng)險(xiǎn)分別是TT基因型個(gè)體的2.05倍(95%CI=0.12-34.63)。但差異無統(tǒng)計(jì)學(xué)意義。肝癌高發(fā)家族(A+B)組中GT基因型的分布頻率明顯高于正常對(duì)照(C)組,但差異有無統(tǒng)計(jì)學(xué)意義(P=0.380.05,OR=2.64,95%CI：0.30-23.35),經(jīng)性別、年齡、吸煙、飲酒、HBsAg等因素校正后患肝癌的風(fēng)險(xiǎn)仍然增高(OR=2.02,95%CI:0.21-19.88,P=0.550.05)。與TT基因型相比,GT基因型在肝癌高發(fā)家族(A+B)組和正常對(duì)照(C)組間的分布頻率差異無統(tǒng)計(jì)學(xué)意義(P=0.550.05)。(4)肝癌組織中,PLXNC1蛋白表達(dá)水平(3.12±1.12)顯著高于肝癌癌旁組織(1.54±0.67)和正常肝組織(1.23±0.87)(P0.05),但PLXNC1蛋白表達(dá)水平在肝癌癌旁組織和正常肝組織間比較無統(tǒng)計(jì)學(xué)差異(P0.05)。(5)肝癌組織中,ITGβ1蛋白表達(dá)水平(3.45±1.25)顯著高于肝癌癌旁組織(1.76±0.98)和正常肝組織(1.02±0.56)(P0.05),在肝癌癌旁組織中ITGβ1蛋白表達(dá)水平顯著高于正常肝組織(P0.05))。(6)肝癌組織中SEMA7A蛋白表達(dá)水平(3.03±1.07)顯著高于肝癌癌旁組織(1.47±0.58)和正常肝組織(1.17±0.92)(P0.05),但SEMA7A蛋白表達(dá)水平在肝癌癌旁組織和正常肝組織間比較無統(tǒng)計(jì)學(xué)差異(P0.05)。(7)肝癌組織中,RAC1蛋白表達(dá)水平(3.25±1.14)顯著高于肝癌癌旁組織(1.68±0.87)和正常肝組織(0.92±0.67)(P0.05),在肝癌癌旁組織中,RAC 1蛋白表達(dá)水平顯著高于正常肝組織(P0.05)。(8)PLXNC1蛋白ITGβ1蛋白、SEMA7A蛋白、RAC1蛋白表達(dá)水平與HCC患者性別、年齡、腫塊大小、HBsAg、AFP間無明顯差異(P0.05)。結(jié)論1、PLXNC1基因rs2272335位點(diǎn)C等位基因型可能是HCC發(fā)生的風(fēng)險(xiǎn)基因型,PLXNC1基因rs2272335位點(diǎn)與廣西肝細(xì)胞癌遺傳易感性顯著相關(guān)ITGβ1基因rs22298141位點(diǎn)多態(tài)性與廣西肝細(xì)胞癌遺傳易感性未發(fā)現(xiàn)顯著相關(guān)；RAC1基因rs6951997位點(diǎn)多態(tài)性與廣西肝細(xì)胞癌遺傳易感性未發(fā)現(xiàn)顯著相關(guān)。2軸突導(dǎo)向通路SEMA7A-ITGβ1-PLXNC1及相關(guān)基因(PLXNC1、JTGβ1、RAC1和SEMA7A)蛋白表達(dá)與廣西肝細(xì)胞的遺傳易感性可能具有相關(guān)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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.7
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本文編號(hào)：2011134