miR-122對(duì)乙型肝炎病毒抗原表達(dá)的影響.pdf

1、目的:探討miR-122對(duì)乙型肝炎病毒HBsAg、HBeAg表達(dá)的影響。方法:以空白對(duì)照組(陰性對(duì)照組),GFP(反義寡核苷酸序列)組為陰性對(duì)照,設(shè)計(jì)合成2條miR-122的反義寡核苷酸序列anti-miR-122,LNA-antimiR-122(反義鎖核苷酸)以及人miR-122的寡核苷酸序列(hsa-miR-122)。脂質(zhì)體包裹后分別以400nm/L(GFP組),400nm/L(anti-miR-122組),40nm/L(LNA-a

2、ntimiR-122組),400nm/L(hsa-miR-122)轉(zhuǎn)染HepG2.2.15細(xì)胞。細(xì)胞轉(zhuǎn)染24小時(shí),48小時(shí)后取培養(yǎng)液上清,時(shí)間分辨熒光免疫法檢測(cè)HBsAg,HBeAg的表達(dá)。轉(zhuǎn)染48小時(shí)后,提取總RNA,Taqman技術(shù)實(shí)時(shí)熒光定量PCR檢測(cè)anti-miR-122組,LNA-antimiR-122組,hsa-miR-122組,GFP組,和陰性對(duì)照組內(nèi)細(xì)胞miR-122的含量。結(jié)果:①轉(zhuǎn)染24小時(shí)后HBsAg表達(dá):hsa

3、-miR-122組HBsAg表達(dá)低于陰性對(duì)照組(p<0.05),has-miR122組HBsAg表達(dá)低于anti-miR-122組(p<0.01),anti-miR122組HBsAg表達(dá)高于GFP組(p<0.01)。②轉(zhuǎn)染24小時(shí)后HBeAg表達(dá):anti-miR122組HBeAg表達(dá)高于陰性對(duì)照組(p<0.01),LNA-antimiR-122組HBeAg表達(dá)顯著高于陰性對(duì)照組(p<0.01),hsa-miR-122組HBeAg表達(dá)低

4、于anti-miR122組(p<0.01),GFP組HBeAg表達(dá)低于anti-miR122組(p<0.01)。③轉(zhuǎn)染48小時(shí)后HBsAg表達(dá):GFP組相對(duì)于陰性對(duì)照組,HBsAg表達(dá)兩者沒(méi)有差異,(p=1.000),anti-miR-122組HBsAg表達(dá)高于陰性對(duì)照組(p<0.05),hsa-miR-122組HBsAg表達(dá)低于anti-miR122組(p<0.01),hsa-miR-122組HBsAg表達(dá)低于LNA-antimiR-

5、122組(p<0.05),anti-miR122組HBsAg表達(dá)高于GFP組(p<0.05)。④轉(zhuǎn)染48小時(shí)后HBeAg表達(dá):LNA-antimiR-122組HBeAg表達(dá)高于陰性對(duì)照組的表達(dá)(p<0.05),hsa-miR-122組HBeAg表達(dá)低于LNA-antimiR-122組(p<0.05),GFP組HBeAg表達(dá)低于LNA-antimiR-122組(p<0.05)。⑤實(shí)驗(yàn)組各組細(xì)胞內(nèi)miR-122的表達(dá):LNA-anti-mi