为寻找放射敏感性基因,采用基因芯片技术筛选出辐射可诱导表达mRNA上调的基因,其中就有IER5.为探索IER5基因的生物学功能及其在宫颈癌放疗中的作用,采用RNA干扰技术构建IER5基因表达抑制的质粒载体并构建IER5-siRNA-HeLa细胞系.对该细胞系与HeLa细胞进行辐射,旨在了解IER5-siRNA-HeLa的细胞生长曲线、细胞周期等实验参数的变化,揭示了IER5基因在辐射诱导中的生物学功能.实验发现,IER5基因表达抑制可提高细胞分裂进入S期与G2-M期的比例,促进细胞分裂,促进细胞生长,提高细胞对辐射的拮抗性,同时发现IER5-siRNA-HeLa细胞在尺寸上大于HeLa细胞.研究表明,IER5基因表达抑制可促使细胞受辐射后发生S期与G2-M期的阻滞,IER5参与辐射细胞周期的调控,对临床宫颈癌放疗有一定的潜在应用价值.
In order to identify the radiosensitivity-related genes, microarray technology was used for candidate genes screening and it was identified that IER5 (Immidiate early response 5) could be up-regulated after radiation. Biological relationship between IER5 and radiation was investigated, and its biological function was examined in human cervical carcinoma treated with radiation. RNAi technology was employed to knock down IER5 gene in HeLa cells and establish the stable transfected cell lines, IER5-siRNA-HeLa. It was found that there was more S/G2/M phases cells with larger size in IER5-siRNA-HeLa cells than that in the control cells when treated with irradiation. The results indicated that IER5 might be involved in cell growth and proliferation. Most importantly, IER5 knockdown increased the radioresistance in cells, which indicated IER5 might be a potential target for the radiotherapeutical treatment in the cervical carcinoma.