【摘要】 目的 探討丙型肝炎病毒非結(jié)構(gòu)蛋白NS4B對肝細(xì)胞內(nèi)p53表達(dá)的影響，以及在肝癌發(fā)生中的作用與機(jī)制。 方法 設(shè)置空白對照組、空白載體組、轉(zhuǎn)染NS4B組、轉(zhuǎn)染p53組、共轉(zhuǎn)染NS4B及p53組。使用脂質(zhì)體介導(dǎo)轉(zhuǎn)染法，轉(zhuǎn)染丙型肝炎病毒非結(jié)構(gòu)蛋白重組質(zhì)粒PCXN2-NS4B及突變型p53基因重組質(zhì)粒pC53-CX22AN3進(jìn)入Chang肝細(xì)胞內(nèi)，并用G418篩選獲得穩(wěn)定表達(dá)細(xì)胞。采用免疫細(xì)胞化學(xué)法檢測p53表達(dá)率。 結(jié)果 空白對照組無p53表達(dá)，空白載體組及轉(zhuǎn)染NS4B組呈弱陽性表達(dá)，轉(zhuǎn)染p53組及共轉(zhuǎn)染組呈陽性表達(dá)；轉(zhuǎn)染p53組、共轉(zhuǎn)染組分別與空白對照組、空白載體組及轉(zhuǎn)染NS4B組比較，差異均有統(tǒng)計學(xué)意義 （P lt;0.05）。 結(jié)論 NS4B可能抑制p53表達(dá)，也可能阻止其進(jìn)入細(xì)胞核，但NS4B與突變型p53關(guān)系不明確。NS4B導(dǎo)致肝細(xì)胞異常增生，誘導(dǎo)肝癌發(fā)生可能不依賴p53的異常表達(dá)及突變。
【Abstract】 Objective To investigate the effect of hepatitis C Virus on-structural protein 4B(HCV NS4B) on expression of p53 in hepatic cell, and to study the role and mechanism in development of hepatocellular carcinoma. Methods The experiment was divided into negative control, pure vector PCXN2, PCXN2-NS4B, PC53-cx22AN3, and co-transfection group. Recombinant plasmid PCXN2-NS4B and mutant p53 gene--PC53-cx22AN3, PC53-cx22AN3 with PCXN2-NS4B, blank vectors were transfected into Chang liver cell by liposome-mediated transfection respectively. Positive cells were screened by G418. The expression rate of p53 was measured by immunocytochemistry. Result No expression rate of p53 gene in control group was found, lower positive expression in group PCXN2 and PCXN2-NS4B. The expression of p53 gene in group PC53-CX22AN3 and co-transfection was ber than the others (P lt;0.005). Conclusion HCV-NS4B may inhibit the expression of p53 gene, and it may play a crucial role in inhibiting p53 transfered to hepatic cells nuclear. But it isn’t clear that the. HCV-NS4B can enhance the role of mutant p53 gene. It suggested that HCV-NS4B induce proliferation of hepatic cell not through regulating the expression of p53.
引用本文： 徐建玉,李昌平,陳楓,陳霞,王忠瓊. 轉(zhuǎn)染丙型肝炎病毒非結(jié)構(gòu)蛋白NS4B對p53表達(dá)的影響. 華西醫(yī)學(xué), 2010, 25(8): 1426-1428. doi: 復(fù)制
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- 2. Muramatsu S, Ishido S, Fujita T, et al. Nuclear localization of the NS3 protein of hepatitis c virus and factors affecting the localization[J]. J Virol, 1997, 71(7): 4954-4961.
- 3. Beerheide W, Tan YJ, Teng E, et al. Downregulation of proapoptotic proteins Bax and Bcl-X(S) in p53 overexpressing hepatocellular carcinomas[J]. Biochem Boophys Res Commun, 2000, 273(1): 54-61.
- 4. 劉重陽, 劉為紋, 陳東風(fēng), 等. 丙型肝炎病毒核心蛋白對細(xì)胞凋亡的影響[J]. 世界華人消化雜志, 2001, 9(10): 1125-1128.