DOC2抑制对宫颈癌SiHa细胞系生长.docVIP

DOC2抑制对宫颈癌SiHa细胞系生长 　 作者：李萍，辛晓燕，刘淑娟，宋庆贺，毛敬 【关键词】 宫颈癌 Inhibitory effect of DOC2 gene on growth of human cervical cancer SiHa cells 　　【Abstract】 AIM: To explore the inhibitory effect of DOC2 gene on the growth of human cervical cancer cell line SiHa. METHODS: Recombinant eukaryotic expression vector pcDNA3.1p93 (containing exogenous human DOC2cDNA) and vector pcDNA3.1 (with neomycin resistance gene only) were transfected into human cervical cancer SiHa cells (no DOC2 gene expression) with lipofectamine respectively. The growth rate, cell cycle and colony formation rate of transfected cells were observed by MTT assay and FCM assay respectively. RESULTS: The growth of SiHa transfected with DOC2 gene was markedly suppressed （Plt;0.05）. Colony formation rate in soft agar was also decreased significantly （Plt;0.05）. Cell cycle analysis showed that the percentage of cells in G1 phase of SiHap93 cells was significiantly increased while that of cells in S phase was decreased. The percentage of cells in the different phases of the cycle did not significantly vary between the SiHa and SiHapcDNA3.1. CONCLUSION: DOC2 gene can inhibit the proliferation and DNA synthesis of human cervical cancer. 　　【Keywords】 cervix neoplasms; DOC2; gene therapy 　　【摘要】 目的： 探讨卵巢癌缺失2(DOC2)基因对宫颈癌SiHa细胞系生长的抑制作用. 方法： 采用基因转染技术，将含有全长DOC2cDNA的真核重组表达质粒和空载体质粒(pcDNA3.1)转染到人宫颈癌SiHa细胞系（无DOC2基因的表达）中，了解其对细胞增殖能力及细胞周期的影响. 结果： 转染DOC2基因的宫颈癌细胞生长受到抑制（Plt;0.05），其在软琼脂克隆形成能力明显降低（Plt;0.05）. DOC2有使G1期细胞比例增高、S期细胞比例下降的趋势,但SiHa细胞和SiHapcDNA3.1细胞之间无显著差异. 结论： DOC2基因能抑制宫颈癌细胞系的增殖. 　　【关键词】 宫颈肿瘤；DOC2；基因疗法 　　0引言 　　卵巢癌缺失2（differentially expressed in ovarian cancer 2, DOC2）基因能抑制多种肿瘤细胞系的生长，是一个新的候选的肿瘤抑制基因. 为探讨DOC2基因对人宫颈癌细胞的抑制作用，我们采用基因转染技术研究其对宫颈癌细胞增殖的影响，为宫颈癌的治疗提供思路. 　　1材料和方法 　　1.1材料无DOC2基因表达的人源性宫颈癌细胞系SiHa由西京医院妇产科实验室提供，常规方法复苏细胞并在37℃, 50 mL/L CO2标准条件下培养. 真核表达载体质粒pCDNA3.1p93(含有人DOC2cDNA)、空载体质粒pCDNA3.1和免疫组化试剂DOC2抗体均由刘淑娟博士惠赠［1-2］. DMEM，质粒转染试剂盒（Lipofectamine 2000）及胰蛋白酶购自GIBCO公司；胎牛血清购自杭州四季青;MTT购自华美生物工程公司； SABC试剂盒与DAB显色剂均购自武汉博士德生物技术有限公司,G418购自Sig