本文選題：多發(fā)性內(nèi)分泌腺瘤2A型 + RET��； 參考：《中南大學(xué)》2012年博士論文

【摘要】：研究背景及目的：多發(fā)性內(nèi)分泌腺瘤2型(multiple endocrine neoplasia type2, MEN2; MIM171400)是一種常染色體顯性遺傳的神經(jīng)內(nèi)分泌腫瘤易感性綜合征,以可變外顯率的甲狀腺髓樣癌(medullary thyroid carcinoma, MTC)、嗜鉻細(xì)胞瘤(pheochromocytoma, PHEO)和甲狀旁腺功能亢進(jìn)(hyperparathyroidism, HPT)為特征。MEN2可分為三種亞型：MEN2A型(multiple endocrine neoplasia type2A; MIM171400)、MEN2B型(multiple endocrine neoplasia type2B; MIM162300)和家族性甲狀腺髓樣癌(familial medullary thyroid carcinoma, FMTC; MIM155240)。 MEN2A臨床上主要表現(xiàn)為甲狀腺髓樣癌、嗜鉻細(xì)胞瘤和原發(fā)性甲狀旁腺功能亢進(jìn),約占MEN2的55%。FMTC一般不涉及其他內(nèi)分泌病變,占MEN2的35%～40%,而MEN2B臨床表現(xiàn)為甲狀腺髓樣癌、嗜鉻細(xì)胞瘤和粘膜神經(jīng)瘤,占MEN2的5%～10%。MEN2通常由RET原癌基因(the rearrangedduring transfection protooncogene gene, RET; MIM164761)或1型神經(jīng)營(yíng)養(yǎng)因子酪氨酸激酶受體基因(the neurotrophic tyrosine kinase receptor type1gene, NTRKI; MIM191315)突變引起。 MEN2是一種起病隱匿,易被延誤診治的疾病。對(duì)家系內(nèi)未發(fā)病的攜帶者進(jìn)行多發(fā)性內(nèi)分泌瘤2型遺傳檢測(cè),可為臨床前診斷和早期治療提供重要幫助。目前認(rèn)為檢測(cè)基因突變是診斷MEN2的金標(biāo)準(zhǔn),并可用于指導(dǎo)臨床選擇治療方案。對(duì)基因功能的更深入研究,將會(huì)使MEN2的基因治療成為可能。為了調(diào)查中國(guó)漢族MEN2A家系中的遺傳致病基因以及基因型和表型之間的關(guān)系,我們對(duì)一個(gè)3代MEN2A家系9名成員(5名患者)進(jìn)行了遺傳學(xué)分析。 方法：收集1個(gè)3代中國(guó)漢族MEN2A家系9名家系成員的外周血,其中包括5名患者。抽取先證者和她妹妹的外周血做淋巴細(xì)胞培養(yǎng),用植物凝集素刺激72小時(shí)后收集細(xì)胞中期分裂相,應(yīng)用染色體核型分析吉姆薩胰蛋白酶顯帶(Giemsa-trypsin-Giemsa, GTG;G顯帶)技術(shù)對(duì)染色體分裂相進(jìn)行分析。對(duì)MEN2A家系先證者進(jìn)行RET基因和NTRK1基因全部編碼區(qū)聚合酶鏈?zhǔn)椒磻?yīng)(Polymerase Chain Reaction, PCR)擴(kuò)增后直接測(cè)序。對(duì)家系成員進(jìn)行突變共分離分析,對(duì)100名正常對(duì)照進(jìn)行RET Cys634Arg位點(diǎn)突變檢測(cè)。 結(jié)果：對(duì)兩名患者的G顯帶染色體核型分析發(fā)現(xiàn)均為正常女性染色體核型(46,XX)。對(duì)RET基因和NTRKI基因的整個(gè)編碼區(qū)序列分析發(fā)現(xiàn)了9個(gè)核苷酸變異,包括5個(gè)RET變異：c.1296GA (Ala432Ala)、c.1900TC (Cys634Arg)、c.2071GA (Gly691Ser)、IVS12+47CT和c.2712CG(Ser904Ser);4個(gè)NTRK1變異：-5GA、c.1187CT (Ser396Leu)、IVS16-4delA和IVS18+6CT。其中RET c.1296GA(Ala432Ala)和c.2712CG (Ser904Ser), NTRK1C.1187CT (Ser396Leu)和IVS18+6CT為新的變異。對(duì)家系成員進(jìn)一步分析只有RET突變Cys634Arg與疾病共分離,而在100個(gè)正常對(duì)照中未見該突變。5名已進(jìn)行外科手術(shù)患者均為Cys634Arg雜合突變攜帶者,一名36歲未受累家系成員為RET Cys634Arg雜合突變攜帶者。 結(jié)論：對(duì)一個(gè)中國(guó)漢族多發(fā)性內(nèi)分泌腺瘤2A型家系進(jìn)行G顯帶染色體核型分析和基因突變分析,發(fā)現(xiàn)了9個(gè)核苷酸變異,包括5個(gè)RET變異：c.1296GA (Ala432Ala)、c.1900TC (Cys634Arg)、 c.2071GA (Gly691Ser)、IVS12+47CT和c.2712CG (Ser904Ser);4個(gè)NTRK1變異：-5GA、c.1187CT (Ser396Leu)、IVS16-4delA和IVS18+6CT。其中RET c.1296GA (Ala432Ala)和C.2712CG (Ser904Ser), NTRK1c.1187CT (Ser396Leu)和IVS18+6CT為新的變異。RET c.1900TC (Cys634Arg)變異是致病突變。一名36歲Cys634Arg攜帶者無臨床癥狀暗示該疾患可能存在外顯不全或發(fā)病延遲。本研究有助于進(jìn)一步了解多發(fā)性內(nèi)分泌腺瘤2A型基因型和表型關(guān)系,同時(shí)對(duì)該家系遺傳咨詢和產(chǎn)前診斷有重要意義。 研究背景及目的：慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD; MIM606963)是一種常見的、發(fā)病率和死亡率逐年上升的復(fù)雜性疾病,發(fā)病機(jī)制主要是多基因變異和環(huán)境因素相互作用。吸煙是COPD最主要的致病因素,不同個(gè)體之間存在COPD易感性差異。更好的理解COPD易感的遺傳因素有助于發(fā)現(xiàn)新的藥物靶點(diǎn),從而促進(jìn)COPD的早期診斷和治療。 人類基因組計(jì)劃和國(guó)際人類基因組單體型圖計(jì)劃(the International HapMap Project)的完成從全基因組水平提供了研究COPD遺傳機(jī)制的平臺(tái)。全基因組關(guān)聯(lián)研究(genome-wide association study, GWAS)能對(duì)整個(gè)基因組成千上萬個(gè)單核苷酸變異(single nucleotide polymorphism, SNP)研究,以確定那些與疾病癥狀同時(shí)出現(xiàn)的SNP。該技術(shù)作為一種前沿和有效的研究復(fù)雜疾病的方法,已經(jīng)應(yīng)用于COPD相關(guān)研究中,并已取得了一些陽(yáng)性結(jié)果,但這些結(jié)果需要在大樣本、多種族的人群中進(jìn)行驗(yàn)證。 最近GWAS發(fā)現(xiàn)在非亞洲人群中一些基因變異與COPD易感性相關(guān),如鐵效應(yīng)元件結(jié)合蛋白基因(the iron-responsive element-binding protein2gene, IREB2; MIM147582)、家族序列相似性13A基因(the family with sequence similarity13member A gene,FAM13A; MIM613299)和X線修復(fù)交叉互補(bǔ)基因5(the X-ray repair cross-complementing protein5gene, XRCC5; MIM194364)。本研究對(duì)中國(guó)大陸漢族人群COPD患者和正常對(duì)照進(jìn)行IREB2rs2568494變異、FAM13A rs2869967變異和XRCC5rs3821104變異DNA序列分析,以確定中國(guó)漢族人群中這些基因變異與COPD表型的相關(guān)性。 方法：對(duì)275名COPD患者和434名性別、年齡和種族匹配的正常對(duì)照進(jìn)行IREB2rs2568494變異、FAM13A rs2869967變異和XRCC rs3821104變異DNA序列PCR擴(kuò)增后直接測(cè)序,應(yīng)用Assign3.5分析軟件、MT Navigator軟件和DNAstar軟件對(duì)測(cè)序結(jié)果進(jìn)行分析。對(duì)正常人群進(jìn)行Hardy-Weinberg平衡測(cè)試,應(yīng)用SPSS16.0對(duì)病例-對(duì)照人群進(jìn)行基因變異與COPD的相關(guān)性分析。 結(jié)果：正常對(duì)照中所有測(cè)試的基因型頻率符合Hardy-Weinberg平衡。FAM13A rs2869967變異基因型和等位基因頻率分布在正常對(duì)照和COPD患者中存在統(tǒng)計(jì)學(xué)意義(基因型分布：X2=6.319,P=0.042；等位基因分布：X2=4.014,P=0.045)；XRCC5rs3821104變異變異基因型和等位基因頻率分布在正常對(duì)照和COPD患者中也存在統(tǒng)計(jì)學(xué)意義(基因型分布：X2=6.062,P=0.048；等位基因分布：X2=5.607,P=0.018),與文獻(xiàn)報(bào)道結(jié)果一致。IREB2rs2568494變異與COPD表型無統(tǒng)計(jì)學(xué)意義(基因型分布：X2=0.590,P=0.744；等位基因分布：X2=0.034,P=0.854)。 結(jié)論：我們發(fā)現(xiàn)FAM13A rs2869967和XRCC5rs3821104這兩個(gè)變異在中國(guó)漢族COPD患者和正常對(duì)照中的基因型和等位基因頻率分布有統(tǒng)計(jì)學(xué)意義。IREB2rs2568494在中國(guó)漢族COPD患者和正常對(duì)照中的基因型和等位基因頻率分布無統(tǒng)計(jì)學(xué)意義,中國(guó)人群在COPD患病遺傳基礎(chǔ)上可能和非亞洲人群存在差異。遺傳風(fēng)險(xiǎn)變異的發(fā)現(xiàn)有助于建立新的COPD預(yù)警和治療方法。
[Abstract]:Background and purpose: multiple endocrine adenoma type 2 (multiple endocrine neoplasia type2, MEN2; MIM171400) is an autosomal dominant neuroendocrine tumor susceptibility syndrome, with variable exostatic thyroid myeloid carcinoma (medullary thyroid carcinoma, MTC), pheochromocytoma (pheochromocytoma, PHEO) and Hyperparathyroidism (hyperparathyroidism, HPT) can be divided into three subtypes: MEN2A (multiple endocrine neoplasia type2A; MIM171400), MEN2B type (multiple endocrine) and familial thyroid medullary carcinoma. The main manifestations are medullary thyroid carcinoma, pheochromocytoma and primary hyperparathyroidism, about MEN2 of 55%.FMTC generally do not involve other endocrine diseases, accounting for 35% to 40% of MEN2, while MEN2B is a clinical manifestation of medullary thyroid carcinoma, pheochromocytoma and mucous neuroma, accounting for 5% to 10%.MEN2 of MEN2, usually by RET proto oncogene (the rear) Rangedduring transfection protooncogene gene, RET; MIM164761) or type 1 neurotrophic factor tyrosine kinase receptor gene (the neurotrophic tyrosine kinase receptor type1gene) mutation.
MEN2 is a disease of insidious onset and easily delayed diagnosis and treatment. Genetic detection of multiple endocrine neoplasia type 2 can provide important help for preclinical diagnosis and early treatment. It is considered that detection of gene mutation is the gold standard for diagnosis of MEN2 and can be used to guide clinical selection treatment. More in-depth study will make the gene therapy for MEN2 possible. In order to investigate the genetic pathogenicity of the MEN2A family in the Chinese Han family and the relationship between genotype and phenotype, we have carried out a genetic analysis of 9 members of the 3 generation MEN2A family (5 patients).
Methods: the peripheral blood of 1 family members of the MEN2A family of 3 generations of Han Chinese was collected, including 5 patients, including 5 patients. The lymphocyte culture of the precursor and her sister's peripheral blood was collected. The metaphase phase of the cell was collected after 72 hours of plant agglutinin stimulation, and the Giemsa-trypsin-Giemsa was analyzed by karyotype. GTG; G ribbon) technique was used to analyze the chromosomal division phase. The RET gene and the NTRK1 gene were amplified by polymerase chain reaction (Polymerase Chain Reaction, PCR) for direct sequencing of the MEN2A family precursor. The mutation co separation of the family members was analyzed and the RET Cys634Arg site mutation was detected in 100 normal controls.
Results: the karyotype analysis of the G banding chromosomes of two patients was found to be normal female chromosome karyotype (46, XX). 9 nucleotide variations, including 5 RET variants, c.1296GA (Ala432Ala), c.1900TC (Cys634Arg), c.2071GA (Gly691Ser), IVS12+47CT, and NTRKI, were found in the sequence analysis of the whole coding region of the RET and NTRKI genes. 4Ser); 4 NTRK1 variants: -5GA, c.1187CT (Ser396Leu), IVS16-4delA, and IVS18+6CT. in which RET c.1296GA (Ala432Ala) and c.2712CG (Ser904Ser) are new variations. All the patients who had undergone surgery were carriers of Cys634Arg heterozygous mutation. A 36 year old unaffected family member was a RET Cys634Arg heterozygous mutation carrier.
Conclusion: the karyotype analysis and gene mutation analysis of the G banding chromosome of a Chinese Han 2A family with multiple endocrine adenomas found 9 nucleotide variations, including 5 RET variations: c.1296GA (Ala432Ala), c.1900TC (Cys634Arg), c.2071GA (Gly691Ser), IVS12+47CT and c.2712CG (Ser904Ser); 4 variations. CT (Ser396Leu), IVS16-4delA and IVS18+6CT. in which RET c.1296GA (Ala432Ala) and C.2712CG (Ser904Ser), NTRK1c.1187CT (Ser396Leu) and IVS18+6CT are new variants are pathogenic mutations. A 36 year old carrier without clinical symptoms suggests that the disease may have extic or delay in the disease. It is helpful for us to further understand the genotype and phenotype relationship of multiple endocrine neoplasia type 2A, and is of great significance for genetic counseling and prenatal diagnosis of this family.
Background and purpose: chronic obstructive pulmonary disease (COPD; MIM606963) is a common complex disease with increasing incidence and mortality year by year. The main pathogenesis is the interaction of multi gene mutation and environmental factors. Smoking smoking is the most important pathogenic factor of COPD, and the survival of different individuals is stored among different individuals. A better understanding of the genetic factors of COPD susceptibility helps to identify new drug targets, thereby promoting early diagnosis and treatment of COPD. COPD is more susceptible to the disease.
The completion of the human genome project and the the International HapMap Project provides a platform for studying the genetic mechanism of COPD from the whole genome level. The whole genome association study (genome-wide association study, GWAS) can make up thousands of single nucleotide variations (single nucleoti) for the whole gene. De polymorphism, SNP) studies have been applied to the research of COPD related studies and have obtained some positive results in the study of the SNP., which is associated with the symptoms of disease, as a frontier and effective method for the study of complex diseases, but these results need to be verified in large sample and multiracial groups.
GWAS recently found that some of the genetic variations in non Asian populations are associated with COPD susceptibility, such as the iron effect element binding protein gene (the iron-responsive element-binding protein2gene, IREB2; MIM147582), and the family sequence similarity 13A gene (the family with) and X-ray repair. The cross complementation gene 5 (the X-ray repair cross-complementing protein5gene, XRCC5; MIM194364). This study conducted IREB2rs2568494 variation, FAM13A rs2869967 variation and XRCC5rs3821104 variant sequence analysis for COPD patients and normal controls in Chinese mainland Han population to determine the genetic variation and phenotype in Chinese Han population. Relevance.
Methods: 275 COPD patients and 434 normal controls of sex, age and race matched with IREB2rs2568494 mutation, FAM13A rs2869967 variation and XRCC rs3821104 variant DNA sequence PCR were amplified and sequenced directly. Assign3.5 analysis software, MT Navigator software and DNAstar soft parts were used to analyze the sequencing results. Rdy-Weinberg balance test and SPSS16.0 were used to analyze the correlation between gene mutation and COPD in case-control population.
Results: the genotype frequencies of all tests in the normal control were consistent with the Hardy-Weinberg balanced.FAM13A rs2869967 variant genotype and the allele frequency distribution in normal controls and COPD patients (genotype distribution: X2=6.319, P=0.042; allele distribution: X2=4.014, P=0.045); and XRCC5rs3821104 variant variant The frequency distribution of genotype and allele was also statistically significant in normal controls and COPD patients (genotype distribution: X2=6.062, P=0.048; allele distribution: X2=5.607, P=0.018). There was no statistical significance between.IREB2rs2568494 variation and COPD phenotypes (genotype distribution: X2=0.590, P=0.744; allele). B: X2=0.034, P=0.854).
Conclusion: we found that the frequency distribution of genotype and allele of the two mutations of FAM13A rs2869967 and XRCC5rs3821104 in Chinese Han COPD patients and normal controls had statistical significance.IREB2rs2568494 in the genotype and allele frequency distribution of COPD patients and normal controls in Chinese Han people, and there was no statistical significance in the genotype and allele frequency distribution in Chinese Han people and normal controls. There may be differences in the genetic basis of COPD and non Asian populations. The discovery of genetic risk variants helps to establish new COPD early warning and treatment methods.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R563.9

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 戚曉平;應(yīng)榮彪;杜振方;張咸寧;馬炬明;;2型多發(fā)性內(nèi)分泌腺瘤的分子遺傳學(xué)研究進(jìn)展[J];中國(guó)優(yōu)生與遺傳雜志;2010年12期

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