针对HPV16 E7基因的RNA干扰对CaSKi细胞生物学特性的影响  

Biological feature changes of HPV16 positive cervical carcinoma cell resulting from expression inhibition of oncogene E7 by specific siRNA

作　　者：姜向阳[1] 刘文康[2] 张镇西[2] 楚雍烈[3] 

机构地区：[1]西安交通大学第三附属医院暨陕西省人民医院妇产科,陕西西安710068 [2]西安交通大学生命科学与技术学院,生物医学信息工程教育部重点实验室,生物医学工程博士后流动站,陕西西安710049 [3]西安交通大学医学院病原生物学系,陕西西安710061

出　　处：《现代肿瘤医学》2009年第6期1029-1031,共3页Journal of Modern Oncology

摘　　要：目的:探讨RNA干扰抑制HPV16 E7基因对宫颈癌细胞系CaSKi细胞生物学特性的影响。方法:用脂质体将pGENESIL-1/E7(PS7)转染CaSKi细胞内,用透射电子显微镜观察细胞形态变化;细胞计数法测定细胞增殖能力的变化;流式细胞仪测定DNA含量来检测细胞是否发生特征性的凋亡;最后比较细胞裸鼠的成瘤能力。结果:透射电镜显示,凋亡细胞体积变小,细胞核染色质浓缩、趋边,细胞膜表面伪足消失,产生凋亡小体。流式细胞仪检测结果表明,RNA干扰24h、48h和72h后,CaSKi/PSN细胞的凋亡率分别为0.21%、0%、1.62%,CaSKi/PS7细胞的凋亡率分别为31.49%、28.95%、31.54%。生长周期停滞于S期;裸鼠荷瘤实验比较研究显示,RNA干扰后的CaSKi/PS7组的瘤结节体积、重量均较CaSKi/PSN瘤细胞显著降低(P<0.05),CaSKi/PS7组的平均抑瘤率为55.4%。结论:本研究为RNA干扰应用于研究宫颈癌发病分子机制、临床治疗、信息分子药物的开发和干预性预防提供了新的资料和方法。Objective：To investigate the effects of RNAi against HPV16 E7 gene on biological features of cervical carcinoma cell C aSKi. Methods： The plasmids pGENESIL- 1/E7 （PS7）expressing small hairpins RNA （shRNA） against E7 in vivo were transfeced into cervical carcinoma cell CaSKi with lipofectamine2000. The morphological changes of transfected cells were observed by transmission electron microscope（TEM）. Cell growth curve was performed to detect the ability of cell proliferation. DNA content by flow cytometry （FCM） were carried out to detect apoptosis. Fi- nally nude mice were inoculated with CaSKi cells to detect the ability of tumor formation. Results： By TEM CaSKi/ PS7 became smaller, cytoplasmic condensation, nucleus edged and loss of pseudopodium on cell surface, at last nucleus broke into apoptotie body following RNAi. The cell cycle stage was blocked at S phase and the percentage of G1 and G2 stage cell decreased dramatically . After 24h ,48h and 72h of RNAi, the apoptosis rates of CaSKi/PSN were 0.21% ,0%, 1.62% respectively and that of CaSKi/PS7 were 31.49% ,28.95% ,31.54% respectively. As the ability of tumor fomation in nude mice as concerned, compared with CaSKi/PSN the volumes and quality of tumor were siganificantly decreased （P 〈 0.05 ）for CaSKi/PS7, in addition the tumor fomation inhibition rate was 55.4%. Conclusion： These findings suggested that inhibition of the expression of oncogene E7 in HPV16 by means of RNAi was effective and provided new materials for prevention and treatment of cervical carcinoma infected by HPV16.

关 键 词：HPV16 CASKI细胞 E7基因 RNA干扰 

分 类 号：R734.2[医药卫生—肿瘤] R392.2[医药卫生—临床医学]