目的:探讨肾透明细胞癌患者线粒体DNA D-loop(mtDNA D-loop)基因变异的特点,以寻找新的判断肾透明细胞癌预后的标志。方法:对2002-08-01-2007-08-31河北医科大学第四医院59例完整随访的肾癌患者采用外周血基因组DNA,运用聚合酶链反应(PCR)对mtDNA D-loop区进行扩增并测序。将mtDNA D-loop区的测序结果与线粒体文库中的Revised Cambridge Reference Sequence(rCRS)比对进行单核苷酸变异性分析。比较基因变异和临床随访资料与肾透明细胞癌预后的潜在关系,生存曲线分析采用Kaplan-Meier方法,组间比较采用Log-rank检验,多因素分析采用Cox比例风险回归模型。结果:影响肾透明细胞癌患者5年生存率的单因素分析显示,≥55岁组患者5年生存率为67.5%(36/59),低于<55岁组的85.0%(23/59),χ2=124.042,P<0.001;男性5年生存率为71.7%(35/59),低于女性的78.5%(24/59),χ2=15.115,P<0.001;高TNM分期组患者5年生存率为72.5%(39/59),低于低TNM分期组的78.4%(20/59),χ2=11.123,P=0.001;肿瘤直径≥5cm患者生存率为68.3%(37/59),低于肿瘤直径<5cm患者的84.6%(22/59),χ2=103.690,P<0.001。mtDNA D-loop区测序结果显示,有12个分布频率>5%的变异位点。262T和262C的5年生存率分别为70.4%和80.7%,差异有统计学意义,χ2=65.145,P<0.001。多因素分析显示,TNM分期高低、肿瘤直径大小、262位点的变异是影响肾透明细胞癌患者预后的独立危险因素,P<0.001。结论:分析肾透明细胞癌患者mtDNA D-loop区的变异可判断肾透明细胞癌患者的预后,其中262T可作为判断肾透明细胞癌患者预后的一个新型、独立的指标。 Objective: To investigate the characteristics of mitochondrial DNA D-loop (mtDNA D-loop) gene mutations in patients with clear cell renal cell carcinoma (RCCC) to find a new marker for judging the prognosis of renal clear cell carcinoma. Methods: A total of 59 patients with renal cell carcinoma from the Fourth Hospital of Hebei Medical University from August 2002 to January 2007 were enrolled in this study. Genomic DNA was extracted from the peripheral blood of patients with renal cell carcinoma and the mtDNA D-loop region was amplified by polymerase chain reaction Increase and sequencing. Single nucleotide variation analysis was performed by aligning the sequencing results of the mtDNA D-loop region with the Revised Cambridge Reference Sequence (rCRS) in the mitochondrial library. The potential relationship between gene mutation and clinical follow-up data and the prognosis of renal clear cell carcinoma was compared. Kaplan-Meier method was used to analyze the survival curves, and Log-rank test was used to compare the two groups. Cox proportional hazards regression model was used to analyze multivariate analysis. Results: The univariate analysis of 5-year survival rate in patients with clear cell renal cell carcinoma showed that the 5-year survival rate of patients ≥55 years old was 67.5% (36/59), which was lower than 85.0% (23/59) , χ2 = 124.042, P <0.001. The 5-year survival rate was 71.7% (35/59) in male and 78.5% (24/59) in female, χ2 = 15.115, P <0.001. The survival rate was 72.5% (39/59), lower than 78.4% (20/59) in low TNM staging group, χ2 = 11.123, P = 0.001. The survival rate was 68.3% (37/59) 84.6% (22/59) of patients with tumors <5 cm in diameter, χ2 = 103.690, P <0.001. The sequencing results of mtDNA D-loop region showed that there were 12 mutation sites with> 5% frequency distribution. The 5-year survival rates of 262T and 262C were 70.4% and 80.7%, respectively, with statistical significance (χ2 = 65.145, P <0.001). Multivariate analysis showed that the TNM staging, tumor size and mutation at site 262 were independent risk factors for the prognosis of patients with renal clear cell carcinoma (P <0.001). Conclusion: The analysis of the variation of mtDNA D-loop region in patients with clear cell renal cell carcinoma can judge the prognosis of patients with clear cell renal cell carcinoma. 262T can be used as a new independent indicator to judge the prognosis of patients with clear cell renal cell carcinoma.