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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。
Objective To mutate the exon12 gene of pot1 (protection of telomeres1) in cultured human cancer cell lines (lines). Methods Chromosomal DNA was extracted from 27 cultured human cancer cell lines (lines). The hpot1 gene exon 12 was amplified by PCR. The purified PCR products were directly sequenced. The sequencing results were compared with the data of pot1 gene in GenBank and NCBI database. Of human cancer cell lines (lines) hpot1 gene exon12 mutation analysis. Results Sequencing results showed that five of the human cervical cancer cell lines Hela and human ovarian cancer cell lines HO8910-PM (female hyperplasia) were derived from four female reproductive cancer cell lines and had the same substitution mutation in exon12 region of hpot1 gene (G17722 → C), a missense mutation corresponding to G1385 → C of the cDNA, and Leu454 → Phe of the polypeptide chain; and no mutation was found in the remaining 23 cancer cell lines (lines) of different origin. The results suggest that hpot1 gene exon12 (17722 bases) is a hot spot mutation region, and this mutation may have female reproductive system tumor specificity.