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目的研究喉癌、癌周正常组织及癌前病变中线粒体DNA(mitochondrial DNA,mtDNA)拷贝数改变情况,及其与临床病理特性的关系。方法选取40例喉癌及8例喉癌癌前病变标本,分离癌及癌周组织,分别提取总DNA。选取mtDNA的CoxⅡ和核DNA的β-actin为目的片断,进行荧光定量PCR扩增,以CoxⅡ/β-actin拷贝数平均比定量分析mtDNA的拷贝数情况,并分析拷贝数改变与临床病理特征的关系。结果喉癌组织、癌周组织及癌前病变中mtDNA平均(CoxⅡ/β-actin)比分别为0.9141、0.8103和0.4538,癌和癌周组织、癌和癌前组织及癌周和癌前组织的mtDNA拷贝数差异均有统计学意义(P=0.048,0.003和0.004),有淋巴结转移和无淋巴结转移的喉癌mtDNA平均(CoxⅡ/β-actin)比分别为1.3089和0.6743,差异有统计学意义(P=0.045),mtDNA的拷贝数随肿瘤分期的增加、分化程度的降低而增加但差异无统计学意义(P=0.486和0.459)。结论喉癌中mtDNA拷贝数显著高于癌周组织及癌前病变组织,喉癌中mtDNA拷贝数随淋巴结转移而增加,mtDNA拷贝数改变可能参与喉癌的发生发展。
Objective To study the changes of mitochondrial DNA (mtDNA) copy number in laryngeal cancer, normal peri-cancerous tissues and precancerous lesions and their relationship with clinicopathological features. Methods 40 specimens of laryngeal carcinoma and 8 specimens of precancerous lesions of laryngeal carcinoma were collected, and the cancer and peri-cancerous tissues were isolated and the total DNA was extracted. Select mtDNA Cox Ⅱ and nuclear DNA β-actin fragments for the purpose of quantitative PCR amplification of Cox Ⅱ / β-actin copy number average quantitative analysis of mtDNA copy number and analysis of copy number changes and clinicopathological features relationship. Results The average mtDNA (CoxⅡ / β-actin) ratios in laryngeal cancer tissues, peri-cancerous tissues and precancerous lesions were 0.9141, 0.8103 and 0.4538, respectively, in cancerous and peri-cancerous tissues, cancerous and precancerous tissues, mtDNA copy number difference was statistically significant (P = 0.048,0.003 and 0.004), with lymph node metastasis and no lymph node metastasis of mtDNA average (Cox Ⅱ / β-actin) ratio was 1.3089 and 0.6743, the difference was statistically significant (P = 0.045). The copy number of mtDNA increased with the increase of tumor stage and differentiation, but the difference was not statistically significant (P = 0.486 and 0.459). Conclusion The mtDNA copy number in laryngeal carcinoma was significantly higher than that in peritumorous tissues and precancerous lesions. The mtDNA copy number in laryngeal carcinoma increased with lymph node metastasis. The alteration of mtDNA copy number may be involved in the development of laryngeal carcinoma.