本文選題：內(nèi)皮素1 + 基因多態(tài)性　； 參考：《中南大學(xué)》2014年博士論文

【摘要】：目的：本研究首次在病例對照研究中探索ET-1的SNP位點(diǎn)和單倍型與兒童及青少年骨肉瘤的化療耐藥風(fēng)險(xiǎn)之間的關(guān)系。為證明ET-1基因的SNP和單倍型與兒童及青少年骨肉瘤化療耐藥的風(fēng)險(xiǎn)變化之間存在相關(guān)關(guān)系,提供了第一手的證據(jù),從而為化療耐藥的兒童及青少年骨肉瘤患者在病理生理學(xué)和治療上提出了全新的觀念。 方法：本研究分三部分；1.用350對年齡、性別、腫瘤位置和發(fā)展階段相匹配的兒童青少年骨肉瘤患者進(jìn)行研究。2.苯酚/氯仿法從白細(xì)胞中分離出基因組DNA,并存儲在400毫升的TE中(10mMTris/HCl和1mM EDTA的溶液(pH8.0)。 ET-1基因單核苷酸多態(tài)性的檢測是通過ABI3700DNA分析儀(Applied Biosystems)上進(jìn)行毛細(xì)管電泳寡核苷酸連接測定。對檢測結(jié)果進(jìn)行Hardy-Weinberg平衡檢驗(yàn)、將rs1800541和rs2070699進(jìn)行配對LD試驗(yàn),計(jì)算使用HaploView程序,并對所得進(jìn)行統(tǒng)計(jì)學(xué)分析；3.將患者手術(shù)切除的骨肉瘤標(biāo)本進(jìn)行POCC,對POCC進(jìn)行傳代培養(yǎng),實(shí)時(shí)定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)測定ET-1mRNA水平,并通過酶聯(lián)免疫吸附試驗(yàn)(ELISA)對POCC上清液中細(xì)胞分泌的ET-1水平進(jìn)行測定,觀察腫瘤壞死百分比。 結(jié)果： 1.發(fā)現(xiàn)在年齡、性別及腫瘤位置和發(fā)展階段沒有顯著的差異(表1)。同時(shí)在病例組和對照組之間,在體重指數(shù)(BMI)、收縮壓、舒張壓、血漿ET-1的水平亦無顯著差異(表1)。 2.在三個(gè)SNPs中,對照組中的rs5370明顯偏離了Hardy-Weinberg平衡。 3.在rs1800541上的G等位基因與新輔助化療的弱反應(yīng)或者化學(xué)耐藥性的風(fēng)險(xiǎn)降低有顯著相關(guān)性(P0.01),而位于rs2070699上的G等位基因與耐藥風(fēng)險(xiǎn)增加相關(guān)的傾向并不明顯,P值位于臨界水平(P=0.06)。 4.rs1800541和rs2070699存在較強(qiáng)LD關(guān)系(病例組,D'=0.940,r2=0.885；對照組,D'=0.925,r2=0.860)。 5.2-SNP單倍型TG在病例組中比對照組的顯著相關(guān)性更常見(P=0.012,校正OR,1.82,95％CI,1.10-5.65),而2-SNP單倍型GT在對照組中比病例組中的顯著相關(guān)性更常見(P=0.009,調(diào)整后的 OR,0.25,95％CI,0.14-0.84)。 6.在化療耐藥兒童及青少年骨肉瘤患者中,骨肉瘤細(xì)胞分泌的ET-1 水平與腫瘤壞死的百分比具有顯著的負(fù)相關(guān)關(guān)系。 結(jié)論： 1.兒童青少年骨肉瘤患者在化療后腫瘤壞死與年齡,性別及腫瘤的位置及階段、體重指數(shù)(BMI)、收縮壓、舒張壓、血漿ET-1無相關(guān)性。 2.rs5370與兒童青少年骨肉瘤化療耐藥的風(fēng)險(xiǎn)無明顯相關(guān)性。 3.rs1800541上的G等位基因與新輔助化療的弱反應(yīng)或者化學(xué)耐藥性的風(fēng)險(xiǎn)降低有顯著相關(guān)性,而位于rs2070699上的G等位基因與耐藥性風(fēng)險(xiǎn)增加有相關(guān)性的傾向不明顯。 4.rs1800541和rs2070699存在強(qiáng)LD關(guān)系。 5.TG單倍型和GT單倍型分別與耐藥性兒科骨肉瘤的增高風(fēng)險(xiǎn)和降低風(fēng)險(xiǎn)有關(guān)系。 6.在兒童及青少年骨肉瘤化療耐藥患者中,骨肉瘤細(xì)胞分泌的ET-1水平與腫瘤壞死的百分比具有顯著的負(fù)相關(guān)關(guān)系。
[Abstract]:Objective: To explore the relationship between the SNP locus and haplotype of ET-1 and the risk of chemotherapeutic resistance in children and adolescents with osteosarcoma for the first time in a case-control study. The first hand evidence is provided to demonstrate the correlation between the risk of SNP and haplotype of ET-1 gene and the risk of chemotherapeutic resistance to osteosarcoma in children and adolescents. For children and adolescents with chemotherapy resistant osteosarcoma, a new concept has been put forward in pathophysiology and treatment.
Methods: This study was divided into three parts. 1. the genomic DNA of children with osteosarcoma matched in age, sex, tumor location and development phase was studied by.2. phenol / chloroform method from white cells, and stored in 400 ml of TE (10mMTris/HCl and 1mM EDTA solution (pH8.0). ET-1 gene single nucleotide polymorphisms. The detection was carried out by the ABI3700DNA analyzer (Applied Biosystems) on the capillary electrophoresis oligonucleotide connection. The results were tested by Hardy-Weinberg balance test, rs1800541 and rs2070699 were paired LD test, HaploView program was used, and the results were statistically analyzed. 3. the osteosarcoma resection of the patients was performed. The specimens were POCC, POCC was cultured, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to determine the level of ET-1mRNA, and the level of ET-1 secreted in POCC supernatant was measured by enzyme linked immunosorbent assay (ELISA), and the percentage of tumor necrosis was observed.
Result錛，

本文編號：1923540