p73基因表达模式对于HeLa细胞萘醌类化疗药物敏感性的影响

The impact of p73 gene expression pattern on HeLa cell's sensitivity to a naphthoquinone analog

背景与目的:p73基因的表达,在肿瘤发生、发展以及治疗转归各方面均具有十分重要的意义。本研究旨在探讨HeLa细胞p73基因表达模式对于该细胞萘醌类化疗药物(2,3-dichloro-5,8-dihydroxy-1,4-naphthoquinone,DDN)敏感性的影响。方法:运用TAp73(transcriptional active p73)特异性反义寡核苷酸干预HeLa细胞内p73基因表达模式,以RT-PCR法验证干预效果,同时观察细胞对于DDN药物敏感性的变化;运用MTT和TUNEL法分别检测在DDN药物作用下细胞活力以及凋亡指数(apoptosis index,AI)的变化情况。结果:TAp73特异性反义寡核苷酸特异性阻遏细胞TAp73的表达、下调TAp73/DNp73(dominant negative p73)表达比,从而明显降低HeLa细胞对于DDN的敏感性,导致该药物抑制细胞增殖以及诱导凋亡效能均显著下降(P均<0.01)。相关性分析表明TAp73/DNp73的表达比与细胞活力呈负相关(r=-0.410,P<0.01);而TAp73/DNp73表达比与AI呈正相关(r=0.635,P=0.019)。结论:细胞内TAp73/DNp73表达比的变化对于调节HeLa细胞增殖以及凋亡具有重要的意义,针对该表达比的人工干预可明显影响HeLa细胞对于DDN药物的敏感性。

Background and purpose：The expression of p73 gene is of great importance in both carcinogenesis and chemotherapy sensitivity.The purpose of this study was to investigate the impact of p73 gene expression pattern on HeLa cell＇s sensitivity to a naphthoquinone analog,termed 2,3-dichloro-5,8–dihydroxy-1,4-naphthoquinone（DDN）.Methods：TAp73-specific antisense oligonucleotide was applied to interfere HeLa cellular p73 gene expression pattern.The effect of antisense oligonucleotide was certified by RT-PCR.Meanwhile,the sensitivity of HeLa to DDN＇s proliferation inhibition and apoptosis induction was monitored.Cell viability and apoptosis index were measured by MTT and TUNEL respectively,after TAp73-specific antisense oligonucleotide and DDN treatment.Results：The antitumor activity of DDN,both growth inhibition and apoptosis induction,could be obviously but partly blocked by the intervention of TAp73-specific antisense oligonucleotide.The cell viability and apoptosis index were closely related to the cellular expression ratio of TAp73/DNp73（r=-0.410,P0.01;r=0.635,P=0.019,respectively）.Conclusion：The expression ratio of TAp73/DNp73 is important for HeLa cell＇s proliferation and apoptosis.Changes in p73 gene expression pattern interferes the sensitivity of HeLa cell to DDN.