摘要
目的研究抗HPV16E6核酶(Ribozyme)对宫颈癌CaSKi细胞生长和端粒酶活性的影响。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和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;细胞生长曲线检测细胞生长的改变;TRAP-Elisa法检测端粒酶活性。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和CaSKi明显降低。CaSKi-R细胞生长速度较CaSKi-P和CaSKi明显减慢(P<0.01);CaSKi,CaSKi-P和CaSKi-R三种细胞的端粒酶活性分别为0.89±0.14,0.90±0.11,0.36±0.06,转染了抗HPV16E6核酶的CaSKi-R的端粒酶活性的抑制率为59.55%。经统计学处理,CaSKi-R的端粒酶活性较CaSKi和CaSKi-P细胞明显下降(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,端粒酶活性较前明显下降。
Objective: To investigate the effects of HPV16 E6-ribozyme on cell growth and telomerase activity in cervical carcinoma cells. Methods: With the method of lipofectin transfection, the anti-HPV16 E6-ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which named CaSKi-R, CaSKi-P respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in the three kinds of cells were detected by northern blot. The growth rates of the CaSKi and transfected cells were examined by cell count. Telomerase activity was measured by TRAP-Elisa. Results: Anti-HPV16 E6-ribozyme can be expressed stably in transfected CaSKi-R cells. Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi and CaSKi-P. The growth rate of CaSKi-R was much slower than that of CaSKi and CaSKi-P(P<0.01). Telomerase activity of CaSKi, CaSKi-P, CaSKi-R was 0.89±0.14, 0.90±0.11, 0.36±0.06. The rate of inhibitor was 59.55%. Compared with CaSKi and CaSKi-P cells, telomerase activity in CaSKi-R cells was decreased (P<0.01). Conclusion: CaSKi-R cells transfected by anti-HPVE6-ribozyme showed growth inhibition and decreased telomerase activity.
出处
《中国医学工程》
2004年第5期8-11,共4页
China Medical Engineering
基金
广东省自然科学基金资助项目(编号:96058)
