抑癌基因TSG101及其剪接變異體在正常人外周血細(xì)胞中的表達(dá)
發(fā)布時(shí)間：2019-04-29 17:40
【摘要】：目的:研究抑癌基因TSG101及其四種剪接變異體在健康女性外周血單個(gè)核細(xì)胞中的表達(dá)規(guī)律及突變,探討該基因在乳腺癌發(fā)生發(fā)展過(guò)程中所扮演的角色。 方法:從健康女性外周血單個(gè)核細(xì)胞中提取總RNA,針對(duì)TSG101基因全長(zhǎng)及其四種剪接變異體(V_A、V_B、V_C和V_D)設(shè)計(jì)5對(duì)特異引物進(jìn)行RT-PCR,PCR產(chǎn)物進(jìn)行瓊脂糖凝膠電泳觀察分析,TSG101基因全長(zhǎng)和四種剪接變異體PCR產(chǎn)物進(jìn)行測(cè)序;并從部分樣本提取基因組DNA,運(yùn)用TSG101基因的第4外顯子特異引物,進(jìn)行PCR,對(duì)其產(chǎn)物進(jìn)行序列測(cè)定。 結(jié)果:1、在37例健康女性外周血中均擴(kuò)增出了TSG101全長(zhǎng)及其四種剪接變異體片段。其中V_A、V_B和V_C的表達(dá)較一致,全長(zhǎng)和V_D的表達(dá)存在差別,部分樣本全長(zhǎng)表達(dá)明顯減弱。2、測(cè)序結(jié)果顯示37例中7例(18.92%)均在TSG101基因第4外顯子(mRNA第798bp處)存在“C→T”雜合性點(diǎn)突變;另30例(81.08%)經(jīng)序列比對(duì)與基因庫(kù)的TSG101mRNA序列100%一致。后來(lái)在DNA水平上也證實(shí)了突變的存在。 結(jié)論:1、在健康女性外周血細(xì)胞中存在TSG101基因的全長(zhǎng)及四種剪接變異體片段。2、TSG101基因在健康女性存在雜合性突變,國(guó)內(nèi)外尚未系統(tǒng)報(bào)道。該突變對(duì)剪接變異體B產(chǎn)生了錯(cuò)義突變。3、TSG101基因的雜合性突變可能提示TSG101基因外顯子內(nèi)存在單核苷酸多態(tài)性。
[Abstract]:Aim: to study the expression and mutation of tumor suppressor gene TSG101 and its four splicing variants in peripheral blood mononuclear cells (PBMC) of healthy women, and to explore the role of the gene in the carcinogenesis and development of breast cancer. Methods: total RNA, was extracted from peripheral blood mononuclear cells of healthy women. Five pairs of specific primers were designed for the full length of TSG101 gene and its four splicing variants (V A, V B, V C and V D) for RT-PCR,. The PCR products were analyzed by agarose gel electrophoresis. The full length of TSG101 gene and the PCR products of four splicing variants were sequenced. Genomic DNA, was extracted from some samples and sequenced by PCR, using specific primers of exon 4 of TSG101 gene. Results: 1. The full length of TSG101 and its four splicing variants were amplified in the peripheral blood of 37 healthy women. Among them, the expression of V _ (A), V _ (B) and V _ (C) were the same, the expression of full length and V _ (?) D were different, and the expression of the whole length of some samples was obviously decreased. 2. The sequencing results showed that 7 of 37 cases (18.92%) had "C / T" heterozygosity point mutation in exon 4 of TSG101 gene (mRNA 798 BP). The other 30 cases (81.08%) were identical with the TSG101mRNA sequence of gene bank by sequence alignment. The mutation was later confirmed at the DNA level. Conclusion: 1. The full length of TSG101 gene and four splicing variants exist in the peripheral blood cells of healthy women. 2. The heterozygosity mutation of TSG101 gene in healthy women has not been reported systematically at home and abroad. This mutation produces missense mutation in splicing variant B. 3. The heterozygosity mutation of TSG101 gene may indicate the existence of single nucleotide polymorphism in exon of TSG101 gene.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R346
[Abstract]:Aim: to study the expression and mutation of tumor suppressor gene TSG101 and its four splicing variants in peripheral blood mononuclear cells (PBMC) of healthy women, and to explore the role of the gene in the carcinogenesis and development of breast cancer. Methods: total RNA, was extracted from peripheral blood mononuclear cells of healthy women. Five pairs of specific primers were designed for the full length of TSG101 gene and its four splicing variants (V A, V B, V C and V D) for RT-PCR,. The PCR products were analyzed by agarose gel electrophoresis. The full length of TSG101 gene and the PCR products of four splicing variants were sequenced. Genomic DNA, was extracted from some samples and sequenced by PCR, using specific primers of exon 4 of TSG101 gene. Results: 1. The full length of TSG101 and its four splicing variants were amplified in the peripheral blood of 37 healthy women. Among them, the expression of V _ (A), V _ (B) and V _ (C) were the same, the expression of full length and V _ (?) D were different, and the expression of the whole length of some samples was obviously decreased. 2. The sequencing results showed that 7 of 37 cases (18.92%) had "C / T" heterozygosity point mutation in exon 4 of TSG101 gene (mRNA 798 BP). The other 30 cases (81.08%) were identical with the TSG101mRNA sequence of gene bank by sequence alignment. The mutation was later confirmed at the DNA level. Conclusion: 1. The full length of TSG101 gene and four splicing variants exist in the peripheral blood cells of healthy women. 2. The heterozygosity mutation of TSG101 gene in healthy women has not been reported systematically at home and abroad. This mutation produces missense mutation in splicing variant B. 3. The heterozygosity mutation of TSG101 gene may indicate the existence of single nucleotide polymorphism in exon of TSG101 gene.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R346
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相關(guān)期刊論文 前6條
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