南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (07): 991-993.

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培养的人癌细胞株(系)pot1基因exon12突变筛选

侯敢； 黄迪南； 姜英华；   

  1. 广东医学院生物化学与分子生物学研究所； 广东医学院生物化学与分子生物学研究所 广东湛江524023； 广东湛江524023；
  • 出版日期:2006-07-20 发布日期:2006-07-20
  • 基金资助:
    广东省自然科学基金(04011397)~~

Human pot1 gene exon12 mutation screening in cultured human carcinoma cell strains (lines)

HOU Gan, HUANG Di-nan, JIANG Ying-hua Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang 524023, China   

  1. 广东医学院生物化学与分子生物学研究所； 广东医学院生物化学与分子生物学研究所 广东湛江524023； 广东湛江524023；
  • Online:2006-07-20 Published:2006-07-20

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摘要： 目的对培养的人癌细胞株(系)中pot1(protectionoftelomeres1)基因exon12进行突变筛选。方法从27株培养的人癌细胞株(系)提取染色体组DNA,用PCR方法扩增hpot1基因exon12,PCR产物纯化后直接测序,测序结果与GenBank和NCBI数据库中pot1基因的数据比较,对培养的人癌细胞株(系)中hpot1基因exon12进行突变分析。结果测序结果显示,人宫颈癌Hela细胞株和人卵巢癌细胞株(高转移)HO8910-PM等5个来源于女性生殖系统癌细胞株中的4个,在hpot1基因exon12区有相同的置换突变(G17722→C),为错义突变,对应于cDNA的G1385→C,多肽链的Leu454→Phe;而其余23个不同来源的癌细胞株(系)均无突变。结论提示hpot1基因exon12(17722碱基)是突变热点区,并且这种突变可能具有女性生殖系统肿瘤特异性。 

Abstract: Objective To screen the exon12 mutation of pot1 gene in cultured human carcinoma cell strains (lines). Methods The chromosomal DNA was extracted from 27 cultured carcinoma cell strains (lines). The exon 12 of pot1 gene was amplified by PCR, and the product was purified and screened. The screening results were compared with the data of GenBank and NCBI and the exon 12 mutations in cultured human carcinoma cell strains (lines) analyzed. Results The exon12 sequence of pot1 could be specifically amplified using the designed primers. Direct sequence analysis of the PCR products after purification showed that 4 of the 5 carcinoma cell lines of the female genital system such as Hela and HO8910-PM cells shared the same transition (G17722→C) in exon12 of human pot1 gene resulting in a conversion of G1385→C in the cDNA and amino acid change of Leu454→Phe in the translated polypeptide. The rest of the 23 cell strains (lines) from different origins showed no such mutation. Conclusion The exon12 (17 722 bp) is a mutant region specific for female genital system tumor.

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