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目的 探讨反义硫代寡核苷酸对 HPV16阳性人宫颈癌细胞恶性增殖的影响 .方法 人工合成 3段 18- mer的反义硫代寡聚脱氧核苷酸 AS- ODN1 - 3.其中 ,AS- ODN1(AE6 ) ,AS- ODN2 (AE7)分别与 HPV16 E6 ,E7开放读框起始码及其侧翼序列互补 ,AS- ODN3为一随机序列 .分别用 3种反义寡核苷酸处理整合有 HPV16基因组的宫颈癌细胞 Si-Ha.通过生长曲线测定、3H- Td R参入试验及软琼脂克隆形成试验 ,观察反义寡核苷酸对 Si Ha细胞恶性增殖的影响 .结果 AE6及 AE7均抑制了 Si Ha细胞的生长速率和 DNA合成 ,但对 HPV16阴性的人宫颈癌细胞 He L a无明显影响 ;经 AE6及 AE7处理后的 Si Ha细胞在软琼脂上克隆形成数减少 ;随机序列对 Si Ha细胞增殖无抑制作用 .结论 通过特异性抑制 E6 ,E7基因的表达 ,反义寡核苷酸对 HPV16阳性宫颈癌细胞恶性增殖有显著抑制作用 .
Objective To investigate the effect of antisense oligonucleotides on the malignant proliferation of human papillomavirus (HPV16) -positive human cervical cancer cells.Methods Three segments of 18-mer antisense oligodeoxynucleotides deoxyribonucleotides AS-ODN1-3 were synthesized.Among them, AS - ODN1 (AE6) and AS-ODN2 (AE7) were respectively complementary to the open reading frame of HPV16 E6 and E7 and their flanking sequences, and AS-ODN3 was a random sequence.The three kinds of antisense oligonucleotides Si-Ha cells with HPV16 genome were detected by growth curve assay, 3H-TdR incorporation assay and soft agar colony formation assay, and the effect of antisense oligonucleotide on malignant proliferation of Si Ha cells was observed.Results AE6 and AE7 Inhibited the growth of Si Ha cells and DNA synthesis, but had no significant effect on Hela of HPV16-negative human cervical cancer cells. The number of Si ha cells treated with AE6 and AE7 decreased on soft agar, Si Ha cells.Conclusion Antisense oligonucleotides can significantly inhibit the malignant proliferation of HPV16-positive cervical cancer cells by specifically inhibiting the expression of E6 and E7 genes.