α 粒子诱发人胎气管永生化纤维细胞恶性转化相关基因的分析

Relative gene in immortalized human fetal tracheal fibroblasts malignantly transformed by alpha particle irradiation

目的探索辐射诱发细胞恶性转化相关基因。方法本实验利用３’端引物ＨＴ１１Ｍ（Ｇ，Ｃ，Ａ）与５’端引物ＨＡＰ１７～２４所配的２４对引物同时对ＳＨＴＦ（ＳＶ４０ｉｍｍｏｒｔａｌｉｚｅｄｈｕｍａｎｆｅｔａｌｔｒａｃｈｅａｌｆｉｂｒｏｂｌａｓｔＳＨＴＦ）细胞和经２３８Ｐｕα粒子照射后能在软琼脂上形成克隆的此细胞（αＳＨＴＦ）的ｍＲＮＡ进行反转录ＰＣＲ扩增，６％测序胶电泳，放射自显影。结果发现有２３条基因片段在αＳＨＴＦ细胞中表达，ＳＨＴＦ细胞中不表达。回收差异的基因片段，随机取其中１２条重新扩增后得到６条扩增片段。其中Ｃ１７－５，Ｃ２３－１两片段经Ｎｏｒｔｈｅｒｎｄｏｔ杂交后，均与αＳＨＴＦ细胞ＲＮＡ有强阳性杂交信号，与ＳＨＴＦ细胞ＲＮＡ无杂交信号。结论本实验证实了ＳＨＴＦ和αＳＨＴＦ细胞的基因表达有差异存在。

Objectives\ To explore the relative novel genes which are associated with radiation induced neoplastic transformation of cells. Mothods\ The experiment was carried out by reverse transcription of mRNA from SHTF and α SHTF cells,amplification of cDNA by PCR using 24 pairs of primers consisting of 3′[HT11M(G,C,A)] and arbitrary 5′ (HAP17～24) primers,tracing with 32 P dATP,running on 6% sequencing gels in adjacent lanes, and then autoradiographing. Results\ Twenty three DNA segments showed expression in α SHTF cells,but not in SHTF cells. By reamplification of 12 differential expression DNA segments of them at random,6 DNA segments were obtained.Northern dots showed that C17 5 and C23 1 had positive responses in α SHTF cells,but negative in SHTF cells. Conclusions\ The research indicates that there is difference in gene expression between SHTF cells and SHTF cells.