Cisplatin sensitivity and mechanisms of anti-HPV16 E6-ribozyme on cervical carcinoma CaSKi cell line

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Objective: The aim of this study was to study the cisplatin sensitizing effect and mechanism of anti-HPV16 E6- ribozyme on cervical carcinoma cell line.Methods: The anti-HPV16E6-ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which named as CaSKi-R, CaSKi-P respectively.E6 mRNA, the sensitivity to cisplatin, apoptosis rates, expression of p53, Bcl-2, Bax and C-myc proteins and mRNA were examined by Northern blot, MTT colorimetric assay, PI/Annexin V stained methods, flow cytometry anslysis and RT-PCR, respectively.Results: E6 mRNA was less in CaSKi-R than in CaSKi.The sensitivity of CaSKi-R cells to cisplatin was 2.28 and 2.21 times than that of CaSKi and CaSKi-P cells.The apoptotic rates in CaSKi, CaSKi-P and CaSKi-R cells was (18.9 ± 3.5)%, (19.7 ± 4.8)% and (40.4 ± 4.5)%.The apoptotic rates was increased in CaSKi-R than that of CaSKi cells treated with cisplatin (P = 0.003).Comapred with CaSKi cell, the expression of p53 (P = 0.000), Bax protein (P = 0.002) was significantly higher and the expression of Bcl-2 protein (P = 0.005), C-myc protein (P = 0.005) was significantly lower in CaSKi-R than that of CaSKi cell treated with cisplatin.Comapred with CaSKi cell, the expression of p53, Bax mRNA in CaSKi-R cell treated with cisplatin increased, while Bcl-2, C-myc mRNA decreased.Conclusion: CaSKi-R cells transfected by anti-HPVE6-ribozyme increased the sensitivity to cisplatin.The increase of sensitivity to cisplatin in CaSKi-R cells may be associated with increasing expression of p53, Bax protein, and decreasing expression of C-myc, Bcl-2 proteins.
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