抗HPV16E6核酶对宫颈癌细胞恶性表型的影响

The Effect of Anti-HPV16 E6-Ribozyme on Malignant Phenotypes of Cervical Carcinoma Cell Line

目的 :研究特异性抗HPV16E6核酶对宫颈癌细胞恶性表型的影响。方法 :以脂质体法将抗HPV16E6核酶、空载体质粒分别导入CaSKi细胞 ,命名为CaSKi R ,CaSKi P细胞。测定CaSKi,CaSKi R ,CaSKi P 3种细胞的生长曲线和软琼脂克隆形成率 ,流式细胞仪检测 3种细胞中HPV16E6,PCNA ,C erbB 2蛋白的表达。将裸鼠分为 3组 ,分别在皮下接种CaSKi,CaSKi R ,CaSKi P细胞 ,检测细胞在裸鼠体内的成瘤能力 ;另取一组裸鼠 ,每只在右侧接种CaSKi细胞 ,左侧接种CaSKi R细胞 ,对比这两种细胞的致瘤性。分析特异性抗HPV16E6核酶对宫颈癌细胞恶性表型的影响。结果 :CaSKi P ,CaSKi细胞生长速率相近 ,CaSKi R细胞的生长速度明显降低。CaSKi R细胞的软琼脂克隆形成率明显低于CaSKi和CaSKi P细胞。与CaSKi细胞相比 ,CaSKi R细胞表达HPV16E6,PCNA ,C erbB 2蛋白明显减少 ,而CaSKi P细胞无此改变。CaSKi和CaSKi P在裸鼠体内的致瘤性无显著差异 ,而CaSKi R的成瘤性显著低于CaSKi。结论 :抗HPV16E6核酶的导入能部分逆转宫颈癌CaSKi细胞株的恶性表型 ,其原因可能在于病毒癌基因E6表达的降低 ,以及由此而引起的PCNA ,C erbB

Objective: To investigate the characteristics of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to investigate the possibility and practicality of ribozyme in treatment of cervical cancer. Methods: The anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected by lipofectin transfection into CaSKi cell, which named as CaSKi R, CaSKi P respectively. The morphology and the soft agar forming ability were studied. The expression of E6, PCNA and C erbB 2 genes was studied through Flow Cytometry. The tumorgenicity of each cell was detected by injecting cells into the nude mice skin. Three groups of nude mice were injected by CaSKi, CaSKi R and CaSKi P cell separately. Another group of mice was injected by CaSKi cell on right side and CaSKi R cell on left side. Results: There is no distinct difference of the morphology and growth rate between CaSKi and CaSKi P, but the growth rate of CaSKi R decreased. The soft agar forming rate of CaSKi P was similar with that of CaSKi cells, while that of CaSKi R was found decreased. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could reduce the expression of E6, PCNA and C erbB 2 genes on CaSKi R cells, while this phenomenon was not found on the CaSKi P cells. The tumorgenicity of CaSKi R in nude mice was decreased compared with CaSKi and CaSKi P cells. Conclusion: Anti HPVE6 rivozyme could partly reverse the malignant phenotypes of CaSKi cells. The reason may be the decrease of E6 gene expression, and the succeeding decrease of the PCNA and C erbB 2 genes′ expression.