抗HPV16E6核酶对宫颈癌CaSKi细胞化疗的影响  

Effect of anti-HPV16 E6-ribozyme on the sensitivity of cervical carcinoma cell line to chemotherapy

作　　者：饶智国[1] 张积仁[1] 郑燕芳[1] 周惠[2] 屈良鹄[2] 

机构地区：[1]第一军医大学珠江医院肿瘤中心,广东广州510282 [2]中山大学生命科学学院真核生物学实验室,广东广州510275

出　　处：《第一军医大学学报》2002年第4期316-319,共4页Journal of First Military Medical University

基　　金：广东省自然科学基金(96058)

摘　　要：目的研究抗HPV16E6核酶对宫颈癌CaSKi细胞化疗敏感性的影响。方法以脂质体法将抗HPV16E6-Ribozyme、空载体质粒分别导入CaSKi细胞,命名为CaSKi-R、CaSKi-P细胞。点杂交检测核酶在细胞中的表达,Northern杂交检测三种细胞中E6基因的表达。用克隆形成试验检测3种细胞对化疗的敏感性,Annexine/PI双标法检测细胞凋亡率。结果点杂交证实核酶能在CaSKi-R细胞中稳定表达,Northern杂交证实CaSKi-R中表达E6较CaSKi-P、CaSKi明显降低。CaSKi-R细胞的生长速率较CaSKi-P、CaSKi明显减慢,泰素作用后相对克隆形成率和凋亡率在3种细胞间亦无差异(P>0.05);顺铂作用后CaSKi-R细胞的相对克隆形成率较CaSKi-P、CaSKi明显下降(P<0.01),而凋亡率明显增加(P<0.01)。结论转染抗HPV16E6-ribozyme的CaSKi-R细胞出现一定程度的生长抑制,且对顺铂的敏感性增加,而对泰素的敏感性没有发生改变。Objective To study the role of anti-HPV16 E6-ribozyme in modulating the sensitivity of cervical carcinoma cell lineto chemotherapy. Methods By way of lipofectin transfection, anti-HPV16E6-ribozyme antibody and empty eukaryotic expres-sion plasmids were respectively transfected into CaSKi cells that were designated as CaSKi-R and CaSKi-P cells accordingly. Ribozyme expression in the transfected cells was subsequently observed with RNA dot blot, and the amount of E6 mRNA ex-pression detected by Northern blotting. Cell count was performed for determining the growth rate of the non-transfected and transfected CaSKi cells and colony formation test was employed to examine the sensitivity of the cells to chemotherapy. PI/Annexin Ⅴstaining was conducted to determine the apoptosis rates of each cell line in response to chemotherapy. Results As shown by dot blot analysis, stable expression of anti-HPV16E6-ribozyme was achieved in CaSKi-R cells, in which lower E6 mRNA expression level was observed by means of Northern blotting in comparison with those in CaSKi-P and CaSKi cells. Significant slow-down of the growth rate occurred in CaSKi-R cells as compared with the growth rate of the other 2 cell lines, but no differences in relative cloning efficiency and apoptosis rates of the 3 cell lines were observed in response to taxol treatment (P>0.05), while cis-platinum treatment produced comparatively deceased relative cloning efficiency and increased apoptosis rate in CaSKi-R (P<0.01). Conclusion Transfection by anti-HPVE6-ribozyme may inhibit the growth of CaSKi cells and increase their sensitivity to cis-platinum but not to taxol.

关 键 词：核酶 人乳头瘤病毒 化疗 药物敏感性 子宫颈肿瘤 

分 类 号：R737.33[医药卫生—肿瘤] R730.53[医药卫生—临床医学]