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目的探讨人癌细胞ｍｔＤＮＡ与正常细胞ｍｔＤＮＡ一级结构的差异。方法采用一步法制备包括肺腺癌ＳＰＣ－Ａ－Ａ、ＰＬＡ－８０１Ｄ、Ａ５４９、Ａ５３１，肝细胞癌ＳＭＭＣ－７７２１，膀胱上皮癌ＥＪ共６个癌细胞系ｍｔＤＮＡ；用ＰｖｕⅡ、ＸｈｏⅠ、ＰｓｔⅠ、ＥｃｏＲⅠ、ＢｓｔＥⅡ、ＨｉｎｄⅢ、ＨｐａⅠ、Ｂｃ１Ⅰ、ＥｃｏＲⅤ、ＳｃａⅠ和ＸｂａⅠ共１１种限制性内切酶对所得ｍｔＤＮＡ进行ＲＦＬＰ分析；并用ＰＣＲ－ＲＦＬＰ方法分析了癌细胞ｍｔＤＮＡ非编码区结构变化。结果６个癌细胞系其ｍｔＤＮＡ基因编码区的３２个酶切位点均无变异，而有３个癌细胞系在其ｍｔＤＮＡ非编码区第１６２７６位核苷酸处出现了ＥｃｏＲⅤ新的酶切位点。结论提示癌细胞ｍｔＤＮＡ基因编码区一级结构相当稳定，而主要的核苷酸变异可能位于其ｍｔＤＮＡ非编码区。
Objective To investigate the differences in primary structure of mtDNA between human cancer cells and normal cells. Methods One-step method was used to prepare mtDNA including lung adenocarcinoma SPC-A-A, PLA-801D, A549, A531, hepatocellular carcinoma SMMC-7721, bladder cancer epithelial carcinoma EJ, and PvuII, XhoI, PstI, EcoRI. A total of 11 restriction endonucleases, BstEII, HindIII, HpaI, Bc1I, EcoRV, ScaI and XbaI were used for RFLP analysis of the mtDNA. The non-coding regions of mtDNA in cancer cells were analyzed by PCR-RFLP. Results There were no mutations in the 32 restriction sites of the mitochondrial DNA coding region of the 6 cancer cell lines, and there were 3 new cancer cleavage sites in the mtDNA non-coding region 16276 of the 3 cancer cell lines. point. The conclusion suggests that the primary structure of the coding region of mtDNA gene in cancer cells is quite stable, and the major nucleotide variation may be located in the non-coding region of mtDNA.