肺癌组织中STR基因座变异规律的研究

发布时间：2018-05-25 13:49

本文选题：法医物证学 + 肺癌　；参考：《重庆医科大学》2017年硕士论文

【摘要】：目的:本研究目的为探索20个常染色体STR基因座及Amel基因座在人肺癌组织中的变异规律,为肺癌组织的个人识别及亲缘鉴定提供依据。方法:1.样本采集:肺癌患者手术切除的癌组织和癌旁正常肺组织,并统计相关临床资料(包含患者性别、年龄、病理分型及分期等)。2.DNA提取:使用组织基因组DNA提取试剂盒分别对肿瘤组织及癌旁正常组织进行DNA提取。3.PCR扩增及分型:使用MicroreaderTM21 Direct ID System人类荧光标记STR复合扩增检验试剂盒对提取的DNA样本进行PCR扩增,然后用3130序列分析仪进行毛细管电泳;采用Gene Mapper ID V3.2基因分析软件进行基因分型。4.统计学分析:通过Excel对基因分型结果进行变异率及变异类型统计,采用SPSS18.0软件对变异分布与肺癌的病理分型、分期及患者的年龄、性别进行chi-square test检验及Cochran-Armitage趋势检验。结果:1.出现变异的次数统计在75例肺癌样本中有24例检出不同于癌旁正常组织的变异,占肺癌组织样本总数的32%。在21个常用STR基因座上共发现55次等位基因变异。D5S818基因座发生变异最多(7次),其次为D3S1358、D12S391基因座(5次),D2S441、Penta E基因座未检出变异。2.出现变异的类型统计在出现等位基因变异的类型中,等位基因增加10次,杂合性等位基因完全丢失10次,部分性丢失35次。并且在同一肺癌组织中多个基因座可同时发生多种变异。3肺癌组织样本的相关临床资料统计结合目前实验结果与肺癌患者的临床资料分析,发现STR位点的变异与肺癌的分型、患者性别无联系(P0.05),与肺癌的分期及患者年龄正相关,各组间比较有统计学差异(P0.05)。结论:肺癌组织中STR基因座较常发生变异,并且变异更可能在年龄大、恶性程度高的肺癌中发生;本实验检出D2S441、Penta E基因座无变异,在今后的研究中可加大样本量,进一步验证可否将其纳入稳定的STR基因座范围,为肿瘤组织等特殊检材的个人识别及亲缘关系鉴定提供依据。
[Abstract]:Objective: to explore the variation of 20 autosomal STR loci and Amel loci in human lung cancer tissues and to provide evidence for individual identification and genetic identification of lung cancer tissues. Method 1: 1. Sample collection: cancer tissues and adjacent normal lung tissues were surgically resected from lung cancer patients, and relevant clinical data were collected (including patient's sex, age, age). Pathological typing and staging. 2. DNA extraction: using tissue genomic DNA extraction kit to extract DNA from tumor tissues and adjacent normal tissues respectively. 3. PCR amplification and typing: MicroreaderTM21 Direct ID System human fluorescent labeled STR amplification test The extracted DNA samples were amplified by PCR with the kit. Then capillary electrophoresis was carried out with 3130 sequence analyzer and genotyping. 4. 4 with Gene Mapper ID V3.2 gene analysis software. Statistical analysis: the variation rate and variation type of genotyping results were analyzed by Excel. Chi-square test and Cochran-Armitage trend test were used to analyze the variation distribution and pathological classification, stage and age and sex of patients with lung cancer by SPSS18.0 software. The result is 1: 1. The frequency of mutation in 75 cases of lung cancer samples 24 cases were different from the adjacent normal tissues, accounting for 32% of the total samples of lung cancer tissue. A total of 55 alleles. D5S818 loci were detected in 21 common STR loci. The most frequent mutations occurred in the alleles of D5S818 loci, followed by D3S1358-D12S391 loci. No mutation was detected in D2S441penta E loci. Statistical analysis of the types of variation in alleles, alleles increased 10 times, heterozygosity alleles lost 10 times, partial alleles lost 35 times. And in the same lung cancer tissues, multiple loci can produce multiple variants at the same time. 3. The relevant clinical data of lung cancer tissue samples. 3. Combined with the current experimental results and the analysis of clinical data of lung cancer patients, we found that the variation of STR loci was associated with the classification of lung cancer. There was a positive correlation between the sex of the patients and the stage of lung cancer and the age of the patients. Conclusion: the mutation of STR locus in lung cancer tissues is more frequent, and the variation is more likely to occur in older lung cancer with high malignancy, and there is no mutation in D2S441penta E locus in this study, which can increase the sample size in future studies. To further verify whether it can be included in the stable STR locus to provide the basis for individual identification and phylogenetic identification of special samples such as tumor tissue.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：D919.4

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