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