全反式维甲酸激活的人肺腺癌细胞基因的筛选

Screening of Genes from Human Lung Adenocarcinoma Cell Line Activated by All-trans Retinoic Acid

目的：筛选人肺腺癌细胞中全反式维甲酸（ａｌｔｒａｎｓ－ｒｅｔｉｎｏｉｃａｃｉｄ，ＡＴＲＡ）激活表达的基因。方法：以我室构建的ＡＴＲＡ诱导前后人肺腺癌细胞系ＧＬＣ－８２的ｃＤＮＡ削减文库各克隆作为探针，与ＡＴＲＡ处理前后ＧＬＣ－８２的ｃＤＮＡ文库Ｌｉｂ．ＧＬＣ－８２，Ｌｉｂ，ＧＬＣ－８２．ＲＡ１，Ｌｉｂ．ＧＬＣ－８２．ＲＡ４进行Ｓｏｕｔｈｅｒｎ杂交，以筛选在ＡＴＲＡ处理后ＧＬＣ－８２ｃＤＮＡ文库中特异存在的ｃＤＮＡ片段。以筛选出的特异存在于ＡＴＲＡ处理后ＧＬＣ－８２ｃＤＮＡ文库中的ｃＤＮＡ片段作为探针，与胎儿各组织总ＲＮＡ进行Ｎｏｒｔｈｅｒ杂交，以检测其在胎儿各组织中的表达情况。结果：其中一克隆ＲＡ９７特异存在于ＡＴＲＡ诱导１天ｃＤＮＡ文库，核酸序列同源分析表明，ＲＡ９７与人２２号染色体长臂１３区－ＰＡＣ克隆ＰＡＣ４０８Ｎ２３同源性很强，其中一段（约６００ｂｐ）与ＰＡＣ４０８Ｎ２３几乎完全同源；此片段与可能的肿瘤抑制基因ＳＮＣ６同源性也相当强，其中一段（约３５０ｂｐ）与ＳＮＣ６几乎完全同源。ＲＡ９７在多种胎儿组织中均有表达，转录本有两个，分别为４．３ｋｂ和４．８ｋｂ。结论：获得一特异存在与ＡＴＲＡ诱导１天ＧＬ?

Objective: To screen the cDNA subtractive libraries of human lung adenocarcinoma cell line GLC-82 treated by all-trans retinoic acid for different durations. Methods: A cDNA fragment RA97 was obtained from the library of one day treatment. Results: Northern blot results showed that RA97 was expressed in different fetal tissues with two transcrips of 4.5kb and 5.1 kb respectively. Homology analysis showed that RA97 was highly homologous with PAC408N23 at human 22q13 and candidate tumor suppressor gene SNC6. Aproximately 600bp and 350bp DNA sequences of RA97 were identical with corresponding regions of PAC408N23 and SNC6 respectively. Conclusion: These results indicate that RA97 probably has some relationship with tumorogenesis.