反义寡核苷酸对HPV16阳性人宫颈癌细胞增殖的影响

Effects of antisense oligonucleotides on the proliferation of cervical cancer cells containing human papillomavirus type 16

目的：探讨反义硫代寡核苷酸对ＨＰＶ１６阳性的人宫颈癌细胞增殖的影响．方法：人工合成三段１８－ｍｅｒ的反义硫代寡核苷酸Ｓ－ＯＤＮ１～３，其中，Ｓ－ＯＤＮ１（ＡＥ６），Ｓ－ＯＤＮ２（ＡＥ７）分别与ＨＰＶ１６Ｅ６，Ｅ７基因的起始码及其下游序列互补，Ｓ－ＯＤＮ３为一随机序列．分别用三种反义寡核苷酸处理整合有ＨＰＶ１６基因组的人宫颈癌细胞ＳｉＨａ．通过四唑盐比色法，３Ｈ－ＴｄＲ参入试验观察不同浓度反义寡核苷酸对ＳｉＨａ细胞增殖的影响．结果：两种反义寡核苷酸ＡＥ６和ＡＥ７均抑制了ＳｉＨａ细胞的增殖及ＤＮＡ合成，且呈现出剂量依赖效应，但对ＨＰＶ１６阴性的人宫颈癌细胞ＨｅＬａ无明显影响．Ｓ－ＯＤＮ３对细胞增殖无抑制作用．结论：人工合成的反义硫代寡核苷酸ＡＥ６和ＡＥ７能够有效抑制ＨＰＶ１６阳性人宫颈癌细胞的增殖．

Aim: To investigate the effects of antisense phosphorothioate oligonucleotides (S ODNs) on the proliferation of cervical cancer cell line SiHa, which harbor HPV16. Methods:Three different antisense oligodeoxynucleotides were used in this study. Two of these, S ODN 1 (AE6) and S ODN 2 (AE7), are complementary to sequences flanking the start codons of HPV16 E6 and E7 open reading frams, respectively. S ODN 3 is a nonsense sequence. SiHa cells were treated with various concentration of oligos. MTT method, 3H TdR incorporation were used. Results: Both AE6 and AE7 markedly inhibited the proliferation and the DNA synthesis of Hela cells which harbor HPV16 but had little effects on SiHa cells that do not. S ODN 3 had none of these effects. Conclusion:Antisense oligodeoxynucleotides can inhibit the proliferation and the DNA synthesis of HPV16 positive human cervical cancer cells.