本文選題：乙型肝炎病毒X蛋白　切入點(diǎn)：miR-221　出處：《重慶醫(yī)科大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：乙型肝炎病毒X蛋白在肝細(xì)胞肝癌（hepatocellular carcinoma,HCC）的發(fā)生發(fā)展過(guò)程中發(fā)揮重要的作用。更重要的是，microRNA221(miR-221)過(guò)表達(dá)可促進(jìn)肝癌的形成，在肝癌的發(fā)生發(fā)展中發(fā)揮重要作用。此外乳腺癌中研究發(fā)現(xiàn)，miR-221可在轉(zhuǎn)錄后水平下調(diào)雌激素受體α(Estrogen Receptor α, ERα)的表達(dá)，而在肝癌中，ERα對(duì)HCC的發(fā)生發(fā)展具有保護(hù)作用。但是我們對(duì)于miR-221及ERα在HBV相關(guān)性肝癌發(fā)生發(fā)展中的作用尚不清楚。因此，本研究旨在探討miR-221在HBV感染相關(guān)性HCC中的促癌功能以及HBx蛋白誘導(dǎo)miR-221上調(diào)促進(jìn)HepG2細(xì)胞異常增殖的分子機(jī)制，為HBV相關(guān)性肝癌的靶向治療提供理論指導(dǎo)。 方法：通過(guò)Adeasy系統(tǒng)構(gòu)建HBx和GFP重組腺病毒載體，HBx腺病毒感染HepG2細(xì)胞后，流式細(xì)胞術(shù)、Western blot、real-time PCR檢測(cè)HBx腺病毒對(duì)細(xì)胞周期、ERα及miR-221的調(diào)控作用。Western blot檢測(cè)HBV陽(yáng)性細(xì)胞及肝癌組織中ERα的表達(dá)情況。miR-221mimic和miR-221inhibitor分別轉(zhuǎn)染HepG2細(xì)胞48h后，流式細(xì)胞術(shù)檢測(cè)細(xì)胞周期變化，Western blot檢測(cè)ERα表達(dá)情況。 結(jié)果：HBx腺病毒感染HepG2細(xì)胞后，可促進(jìn)HepG2細(xì)胞由G1期向S期進(jìn)展，，此外，HBx腺病毒可下調(diào)ERα蛋白、上調(diào)miR-221。MiR-221mimic可促進(jìn)HepG2細(xì)胞增殖，下調(diào)ERα蛋白的表達(dá)，而miR-221inhibitor則抑制細(xì)胞增殖并上調(diào)ERα蛋白的表達(dá)。 結(jié)論：HBx可能通過(guò)上調(diào)miR-221進(jìn)而下調(diào)ERα對(duì)肝癌的保護(hù)性效應(yīng)而促進(jìn)肝癌細(xì)胞異常增殖，靶向miR-221的策略具有抑制肝癌細(xì)胞增殖的治療潛能。
[Abstract]:Objective: hepatitis B virus X protein plays an important role in the occurrence and development of hepatocellular carcinoma (HCC), and the overexpression of microRNA221miR-221) can promote the formation of hepatocellular carcinoma. In addition, the expression of estrogen receptor 偽 -strogen Receptor 偽 (ER 偽) was down-regulated at post-transcriptional level in breast cancer. However, the role of miR-221 and ER 偽 in the occurrence and development of HBV associated hepatocellular carcinoma is not clear. The purpose of this study was to investigate the carcinogenic function of miR-221 in HBV infective associated HCC and the molecular mechanism of miR-221 up-regulation induced by HBx protein to promote the abnormal proliferation of HepG2 cells, and to provide theoretical guidance for the targeted therapy of HBV associated hepatocellular carcinoma. Methods: HBx and GFP recombinant adenovirus vectors were constructed by Adeasy system and infected with HepG2 cells. The regulation of HBx adenovirus on ER 偽 and miR-221 was detected by flow cytometry. Western blot was used to detect the expression of ER 偽 in HBV positive cells and hepatocellular carcinoma tissues. MiR-221 mimic and miR-221inhibitor were transfected into HepG2 cells for 48 h, respectively. Cell cycle changes were detected by flow cytometry and ER 偽 expression was detected by Western blot. Results after infection with HepG2 cell line, HepG2 cells were infected with 10% HBX adenovirus, which could promote the progression of HepG2 cells from G1 phase to S phase. In addition, adenovirus could down-regulate ER 偽 protein, up-regulate HepG2 cell proliferation and down-regulate ER 偽 protein expression. MiR-221inhibitor inhibited cell proliferation and up-regulated the expression of ER 偽 protein. Conclusion the strategy of targeting miR-221 may promote the abnormal proliferation of hepatoma cells by up-regulating miR-221 and down-regulating the protective effect of ER 偽 on HCC. The strategy of targeting miR-221 has the therapeutic potential of inhibiting the proliferation of HCC cells.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R512.62

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