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目的探讨雄激素和雄激素受体(AR)对肝癌细胞株PEG10表达的调控作用。方法设计合成针对人ARsiRNA,并转染HepG2和7404肝癌细胞株。用双氢睾丸酮(DHT)干预HepG2细胞。WesternBlot检测AR和PEG10表达水平。结果从3对ARsiRNA中筛选到1对siRNA(ARsiRNA3),它在2种肝癌细胞株中均可有效抑制AR的表达,其抑制作用呈剂量依赖关系。2种肝癌细胞株中,浓度为240nmolL的ARsiRNA3在转染后24h,对AR抑制效率可达80%以上,且抑制效果可持续至72h。ARsiRNA3转染24h后PEG10表达水平降低,转染48h后,PEG10表达水平降低非常明显,72h后PEG10表达有所上升。DHT可促进HepG2细胞PEG10的表达,呈剂量依赖关系。DHT对AR表达未见明显作用。结论雄激素和AR参与了肝癌细胞株PEG10表达的调控。这可能是男性肝细胞癌发病率较高的原因之一。
Objective To investigate the regulatory effect of androgen and androgen receptor (AR) on the expression of PEG10 in hepatocellular carcinoma cell line. Methods Human ARsiRNAs were designed and synthesized and transfected into HepG2 and 7404 hepatoma cell lines. HepG2 cells were treated with dihydrotestosterone (DHT). Western Blot detection AR and PEG10 expression levels. Results A total of 1 siRNA (ARsiRNA3) was screened from 3 pairs of ARsiRNAs. ARsiRNA3 was able to inhibit the expression of AR in both hepatocellular carcinoma cell lines in a dose - dependent manner. Among the two kinds of hepatocellular carcinoma cell lines, ARsiRNA3 at a concentration of 240 nmolL could inhibit the AR more than 80% at 24h after transfection, and the inhibitory effect could be prolonged to 72h. The expression of PEG10 was decreased after ARsiRNA3 transfection for 24h, and the expression of PEG10 was significantly decreased 48h after transfection. The expression of PEG10 increased after 72h. DHT can promote the expression of PEG10 in HepG2 cells in a dose-dependent manner. DHT had no significant effect on AR expression. Conclusion Androgen and AR are involved in the regulation of PEG10 expression in hepatoma cell line. This may be one of the reasons for the higher incidence of hepatocellular carcinoma in men.