特异性反义寡核苷酸对含HPV16的SiHa细胞的作用

Effects of type-specific antisense oligonucleotides on cervical carcinoma cell line SiHa containing papillomavirus type 16

目的 研究特异性反义寡核甘酸对含 HPV16的 Si-Ha细胞的抑制作用 .方法 人工合成 2条分别与 HPV 16E6、E7基因起始码互补的反义硫代磷酸化核甘酸及 1条随机序列 ,用此 3种反义寡核甘酸处理 Si Ha细胞 ,观察其细胞成活率和细胞蛋白总量的改变 .结果 细胞的生长抑制与反义寡核甘酸的量及持续时间呈正相关 ,Anti E6作用 48h和 72h后细胞蛋白半数改变量 (ID5 0 )的浓度分别为 17μmol· L- 1和 9.4μmol· L- 1左右 ,Anti E7的 ID5 0的浓度 18μmol· L- 1和 10 .2 μmol· L- 1左右 .对照细胞组及随机序列反义寡核甘酸无抑制作用 .结论 此 Anti E6 ,Anti E7能有效地抑制含HPV16的 Si

AIM To investigate the effects of antisense oligonucleotides on cervical carcinoma cell line SiHa containing human papillomavirus type 16. METHODS Three different antisense oligonucleotides were used in this study. Two of the antisense oligonucleotides were complementary to 18 base pair sequences flanking the start codon of the E6 and E7 genes of HPV 16 with a phosphorolation substitution at each base. Control olignucleotide consisted of the same nucleotides in random sequence. Cervical carcinoma cell line SiHa containing HPV 16 and cervical carcinoma cell line HeLa without HPV 16 was treated with various concentrations of antisense oligonucleotides. The rate of surviving cells and the total protein content were determined. RESULTS The growth of SiHa cells was inhibited by the antisense olgonucleotides in a dose and time dependant manner. The ID50 values of anti E6 in SiHa cells after the incubation for 48 h and 72 h were around 17 μmol·L -1 and 9.4 μmol·L -1 respectively. Their value of anti E7 in these cells after the incubation for 48 h and 72 h were about 18 μmol·L -1 and 10.2 μmol·L -1 respectively. The HeLa cells were not affected. These antisense olgonucleotides inhibited the growth. CONCLUSION Growth of the HPV 16 containing SiHa cells is inhibited by the antisense olgonucleotides and not the olionucleotide with random sequence.