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从未用过抗癌细胞毒药物的非小细胞肺癌(NSCLC)患者的手术标本(鳞状上皮癌)提取癌细胞基因组总DNA。对小鼠成纤维(NIH/3T3)细胞行转染实验。获二轮转化细胞,发现二轮转化率是一轮的2.7倍。在转染过程中转化灶出现的多少,与所用DNA的量有一定关系。 二轮转化细胞能在软琼脂上存活生长,接种裸鼠能长出肿瘤,分离肿瘤组织细胞,体外培养传代存活。表明该二轮转化细胞具有肿瘤细胞的特性。 取一轮、二轮转化细胞和裸鼠肿瘤细胞的DNA分别与放射性~(32)P标记的人体特有的Alu重复序列和ras家族基因探针进行Southern印迹转移和分子杂交。结果在三者细胞的DNA中都见有与Alu杂交的条带。这表明在转染过 程中人体特有的Alu重复序列已整合到转化细胞的基因组中。并确定了转化细胞中的转化基因之一的属性为Ha-ras癌基因。本工作提示吸烟可能是人肺鳞癌发生和Ha-ras活化的重要因素。
Surgical specimens (squamous cell carcinoma) of patients with non-small cell lung cancer (NSCLC) who have never used an anticancer drug have extracted cancer cell genome DNA. Transfection experiments were performed on mouse fibroblast (NIH/3T3) cells. Two rounds of transformed cells were obtained and it was found that the second-round conversion rate was 2.7 times that of the round. How much the conversion foci appear during transfection has a certain relationship with the amount of DNA used. The second round of transformed cells can survive on soft agar. Inoculation of nude mice can grow tumors, isolate tumor cells, survive in vitro cultures. This indicates that the second round of transformed cells has the characteristics of tumor cells. The DNA of one and two rounds of transformed cells and tumor cells from nude mice were subjected to Southern blot transfer and molecular hybridization with radioactive 32 P-labeled human-specific Alu repeats and ras family gene probes, respectively. As a result, a band that hybridized with Alu was observed in the DNA of the three cells. This indicates that the human-specific Alu repeat sequence has been integrated into the genome of transformed cells during transfection. And it was confirmed that the property of one of the transformed genes in the transformed cells is the Ha-ras oncogene. This work suggests that smoking may be an important factor in the development of human lung squamous cell carcinoma and Ha-ras activation.