本文關(guān)鍵詞： microRNA 乙肝病毒 復(fù)制 RNA干擾　出處：《中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版)》2017年04期 　論文類型：期刊論文

【摘要】：【目的】探討靶向乙型肝炎病毒的外源性microRNA(amiRNA)的構(gòu)建及其抑制HBV復(fù)制的作用。【方法】應(yīng)用Invitrogen公司miRNA在線設(shè)計(jì)軟件(BLOCK-iT~(TM) RNAi Designer)針對(duì)HBV基因設(shè)計(jì)4條amiRNA序列;構(gòu)建amiRNA真核表達(dá)質(zhì)粒,Lipofectamine2000轉(zhuǎn)染至人肝癌細(xì)胞株HepG2.2.15;熒光定量PCR檢測(cè)HBV-DNA,ELISA檢測(cè)HBsAg和HBeAg表達(dá)水平。【結(jié)果】設(shè)計(jì)合成的4條amiRNA均可下調(diào)培養(yǎng)體系中HBV-DNA水平以及HBsAg和HBeAg水平,與陰性對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P0.05)。其中以miR3抑制HBV-DNA復(fù)制的效果最為顯著�！窘Y(jié)論】靶向HBV基因適當(dāng)位點(diǎn)設(shè)計(jì)合成的amiRNA在體外可以有效抑制HBV的復(fù)制。
[Abstract]:[objective] to investigate the construction of exogenous microRNAi RNAs targeting hepatitis B virus (HBV) and its inhibitory effect on HBV replication. [methods] four amiRNA sequences were designed for HBV gene by Invitrogen miRNA online design software (BLOCK-iTTM) RNAi designer. AmiRNA eukaryotic expression plasmid was constructed and transfected into human hepatoma cell line HepG2.2.15.The expression levels of HBsAg and HBeAg were detected by fluorescence quantitative PCR. [results] the four amiRNA fragments designed and synthesized could down-regulate the levels of HBV-DNA, HBsAg and HBeAg in the culture system. Compared with the negative control group, the difference was statistically significant (P 0.05). The inhibition of HBV-DNA replication by miR3 was the most significant. [conclusion] amiRNA targeting at the appropriate site of HBV gene can effectively inhibit the replication of HBV in vitro.
【作者單位】： 廣東藥科大學(xué)臨床醫(yī)學(xué)院;廣東藥科大學(xué)生命科學(xué)與生物制藥學(xué)院;
【基金】：國家自然科學(xué)基金(81000923) 廣東省科技計(jì)劃項(xiàng)目(2013B021800089,2016A02021602,2014A020212468)
【分類號(hào)】：R373.21
【正文快照】： 慢性乙型肝炎病毒(hepatitis B virus,HBV)感染是導(dǎo)致肝硬化、重型肝炎及肝細(xì)胞癌的主要原因。在中國,有大約9%的人口存在慢性HBV感染。干擾素和核苷(酸)類似物等抗病毒治療藥物仍然是目前治療慢性HBV的唯一選擇[1]。但上述藥物副作用大,不能徹底清除肝細(xì)胞核內(nèi)的共價(jià)閉合環(huán)狀

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 郜玉峰;余莉;葉s，

本文編號(hào)：1491543