發(fā)布時間：2018-02-26 17:00

  本文關(guān)鍵詞： 乙肝病毒 HepG2.2.15 NLRC5 主要組織相容性復(fù)合物1　出處：《瀘州醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過對核苷酸結(jié)合寡聚化結(jié)構(gòu)域樣受體家族（Nucleotide-binding oligomerization domain receptors,NOD-like receptors, NLRs）中C5（Caspase recruitment domain5,CARD5）型受體在穩(wěn)定轉(zhuǎn)染HBV人肝癌細(xì)胞株（HepG2.2.15）中的表達(dá)調(diào)控，觀察該細(xì)胞表面分子主要組織相容性復(fù)合體1（major histocompatibility complex1，MHC-I）的表達(dá)。方法：1.觀察NLRC5和MHC-I在HepG2、HepG2.2.15細(xì)胞株的表達(dá)情況；通過質(zhì)粒轉(zhuǎn)染HepG2.2.15過表達(dá)NLRC5，，觀察對MHC-I的表達(dá)調(diào)控。采用反轉(zhuǎn)錄聚合酶鏈鎖反應(yīng)（reverse transcription-polymerase chain reaction，RT-PCR）,免疫印跡試驗（Western-blot）及流式細(xì)胞術(shù)檢測NLRC5和MHC-Ⅰ的基因和蛋白表達(dá)水平。2.實驗分組，按照細(xì)胞及干預(yù)因子不同分為7組： A組為僅培養(yǎng)HepaG2細(xì)胞（空白對照組）；B組為HepaG2+IFN-γ組；C組為僅培養(yǎng)HepaG2.2.15； D組為HepaG2.2.15+IFN-γ組； E組為HepaG2.2.15+pcDNA3.1-NLRC5；F組為HepaG2.2.15+pcDNA3.1；G組為HepaG2.2.15+pcDNA3.1-NLRC5+來霉素B（LMB）。結(jié)果：1. NLRC5和MHC-Ⅰ在HepaG2.2.15細(xì)胞中，其基因和蛋白水平表達(dá)較HepaG2明顯降低（P0.05）；2.在HepaG2.2.15細(xì)胞中，轉(zhuǎn)染質(zhì)粒過表達(dá)NLRC5后，可上調(diào)MHC-Ⅰ的表達(dá)；3. IFN-γ可誘導(dǎo)NLRC5在HepG2、HepG2.2.15細(xì)胞株的表達(dá)。結(jié)論：1.HBV感染能夠明顯下調(diào)HepG2細(xì)胞NLRC5及MHC-Ⅰ表達(dá)；2.在HepG2.2.15細(xì)胞中，NLRC5可正調(diào)控MHC-Ⅰ的表達(dá)；3. IFN-γ可誘導(dǎo)NLRC5在HepG2、HepG2.2.15細(xì)胞株的表達(dá)。
[Abstract]:Aim: to regulate the expression of nucleotide-binding oligomerization domain receptor (NLRs) in nucleleotide-binding oligomerization domain receptor (NLRs) in stable transfection of HBV human hepatocellular carcinoma cell line HepG2.2.15. The expression of major histocompatibility complex1MHC-I) on the cell surface was observed. Methods: 1. The expression of NLRC5 and MHC-I in HepG2.2.15 cell line was observed. The expression and regulation of HepG2.2.15 were observed by plasmid transfection with HepG2.2.15. Reverse transcription-polymerase chain reactionation was used to detect the gene and protein expression levels of NLRC5 and MHC- 鈪

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