Tian Lv, He Song, Li Xuan, Hu Wen-yan, Peng Pai-lan, Wang Feng, Gao Chang-yi, Ren Hong, Tang Kai-fu
Department of Gastroenterology, Chongqing Medical University, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2011 Jan;19(1):44-7. doi: 10.3760/cma.j.issn.1007-3418.2011.01.012.
To evaluate the inhibitory effects of long antisense RNA on HBV replication in HepG2.2.15 cells. The coding region of HBV S gene was cloned into pTARGET vector in sense and antisense orientations and the recombinant plasmids were transfected into HepG2.2.15 cells which were divided into HBS2 (antisense RNA) group, HBS4 (sense RNA) group and control group. HBsAg and HBeAg in the culture supernate were detected by ELISA. The HBV DNA in the supernate was quantified by real-time PCR. After treatment, the levels of HBsAg in HepG2.2.15 cell supernatants of three groups were 0.621+/-0.027, 3.399+/-0.018 and 2.232+/-0.187 respectively; the levels of HBeAg were 0.749+/-0.019, 1.548+/-0.025 and 1.570+/-0.044 respectively and the levels of HBV DNA were 1.597+/-0.082, 3.381+/-0.297 and 3.610+/-0.063 respectively. The expressions of HBsAg and HBeAg and the HBV DNA level in HBS2 group were remarkably reduced as compared to the control (Z = -2.309, P value is less than 0.05); whereas the sense plasmid transfection (HBS4) did not affect HBeAg (Z = -0.866) and HBV DNA (Z = -1.155) levels in the culture supernate but slightly increased the HBsAg level (Z = -2.309). Antisense RNA might be a useful tool to repress HBV replication.
评估长链反义RNA对HepG2.2.15细胞中HBV复制的抑制作用。将HBV S基因的编码区以正义和反义方向克隆到pTARGET载体中,并将重组质粒转染到HepG2.2.15细胞中,这些细胞被分为HBS2(反义RNA)组、HBS4(正义RNA)组和对照组。通过ELISA检测培养上清液中的HBsAg和HBeAg。通过实时PCR对上清液中的HBV DNA进行定量。处理后,三组HepG2.2.15细胞上清液中HBsAg水平分别为0.621±0.027、3.399±0.018和2.232±0.187;HBeAg水平分别为0.749±0.019、1.548±0.025和1.570±0.044;HBV DNA水平分别为1.597±0.082、3.381±0.297和3.610±0.063。与对照组相比,HBS2组中HBsAg和HBeAg的表达以及HBV DNA水平显著降低(Z = -2.309,P值小于0.05);而正义质粒转染(HBS4)不影响培养上清液中HBeAg(Z = -0.866)和HBV DNA(Z = -1.155)水平,但略微增加了HBsAg水平(Z = -2.309)。反义RNA可能是抑制HBV复制的有用工具。