摘要
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.
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 Northem 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.
