人肺癌基因组DNA对NIH/3T3细胞的转化作用  

THE TRANSFORMATION OF GENOMIC DNA FROM HU- MAN PRIMARY LUNG CARCINOMA ON NIH/3T3 CELLS

作　　者：刘祥麟[1] 马洁羽[1] 孔颂阳[1] 朱家光[1] 李英[1] 周筱梅[2] 茅幼霞 钱莲芳[2] 张予廉 顾健人[2] 

机构地区：[1]浙江医科大学肿瘤基因组,杭州310006 [2]上海肿瘤研究所生化与分子生物学室,上海210032

出　　处：《细胞生物学杂志》1996年第1期28-34,共7页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金;省卫生厅资助项目

摘　　要：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。High molecular weight genomic total DNA was extracted from the surgical samples of patients with human non-small cell lung carcinoma (NSCLC) with the pathological report of squa- mous cell carcinoma which were not treated by chemotherapy. The two round transfection tests were carried out with NIH/3 T 3 cell system. The second round transformation was 2.7 times as efficient as that of the first one. It seemed that the number of transforming focus increase with the amount of DNA used. The second round transforming cells were implanted into nude mice (BALB/c). The growing tumor cells were separated and cultured alive. It was showed that the cells possess the character of tumor cell. The DNA of the primary and secondary transformed cells were analysed by Southern blot with 32p labeled human Alu DNA sequence and ras family gene as probes. The findings indicated that the positive bands hybridizing with human Alu sequence were present in DNA extracted from both transformed and the tumor cells from nude mice. It was identified that human lung carcinoma genomic DNA was integrated into these cells. And Ha-ras gene was considered as the putative transforming sequence in this test.

关 键 词：肺肿瘤 癌 基因组 DNA 成纤维细胞 细胞转化 

分 类 号：R734.202[医药卫生—肿瘤]