本文選題：原發(fā)性肝細(xì)胞癌 + HBV��； 參考：《廣西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:探討ADAMTS-18基因?qū)Ω伟┘?xì)胞系MHCC-97H、Hep G2體外增殖和侵襲能力的影響。方法:收集2016年經(jīng)廣西醫(yī)科大學(xué)第一附屬醫(yī)院肝膽外科手術(shù)治療和病理診斷為HBV陽(yáng)性原發(fā)性肝細(xì)胞癌的癌組織與癌旁組織標(biāo)本共62對(duì)。(1)用Trizol法分別提取癌與癌旁組織總RNA,分別利用q RT-PCR和免疫組化染色技術(shù)檢驗(yàn)癌組織與癌旁組織中ADAMTS-18的m RNA和蛋白的表達(dá)水平差異,并結(jié)合患者臨床病理特征做相關(guān)性分析。(2)提取MHCC-97H、Hep G2和HL-7702細(xì)胞系的總RNA,利用q RT-PCR檢測(cè)細(xì)胞系中ADAMTS-18基因的m RNA表達(dá)量。(3)應(yīng)用慢病毒介導(dǎo)的基因沉默和過(guò)表達(dá)技術(shù)分別構(gòu)建MHCC-97H、Hep G2、HL-7702細(xì)胞系的ADAMTS-18基因沉默和表達(dá)穩(wěn)轉(zhuǎn)細(xì)胞株,并通過(guò)q RT-PCR和Western blot檢測(cè)沉默和過(guò)表達(dá)效率。(4)應(yīng)用MTT法、細(xì)胞劃痕實(shí)驗(yàn)和Transwell小室體外侵襲實(shí)驗(yàn)分別檢測(cè)ADAMTS-18基因沉默和過(guò)表達(dá)前后肝癌細(xì)胞系的體外增殖和遷移侵襲能力的變化。結(jié)果:(1)在HCC癌組織中的ADAMTS-18 m RNA水平和蛋白表達(dá)水平均顯著高于癌旁組織(P0.05);(2)ADAMTS-18 m RNA在HL-7702、MHCC-97H、Hep G2、SMMC-7721、BEL-7404和QGY-7703細(xì)胞系中均有表達(dá)。(3)成功構(gòu)建ADAMTS-18沉默和過(guò)表達(dá)的穩(wěn)轉(zhuǎn)細(xì)胞株MHCC-97H、Hep G2、HL-7702。(4)細(xì)胞劃痕實(shí)驗(yàn)和Transwell小室侵襲實(shí)驗(yàn)結(jié)果顯示,上調(diào)ADAMTS-18的表達(dá)后,MHCC-97H、Hep G2的侵襲能力顯著提高(P0.05);在下調(diào)ADAMTS-18的表達(dá)后,MHCC-97H的侵襲能力明顯降低(P0.05)。(5)MTT方法檢測(cè)結(jié)果發(fā)現(xiàn),上調(diào)或下調(diào)ADAMTS-18的表達(dá)后,MHCC-97H、Hep G2、HL-7702細(xì)胞系的增殖能力隨之明顯升高和下降(P0.05)。結(jié)論:1、ADAMTS-18基因在HBV相關(guān)性HCC癌組織中的表達(dá)水平顯著高于癌旁組織;2、上調(diào)ADAMTS-18基因的表達(dá)可促進(jìn)MHCC-97H的體外遷移侵襲能力;下調(diào)ADAMTS-18基因的表達(dá)能抑制MHCC-97H的體外遷移侵襲能力;3、上調(diào)ADAMTS-18基因可促進(jìn)MHCC-97H、Hep G2和HL-7702細(xì)胞系的增殖能力。下調(diào)ADAMTS-18基因可抑制上述細(xì)胞系的增值能力。
[Abstract]:Aim: to investigate the effect of ADAMTS-18 gene on the proliferation and invasion of hepatocellular carcinoma cell line MHCC-97 Hep G2 in vitro. Methods: a total of 62 pairs of carcinomas and paracancerous tissues were collected from the primary hepatocellular carcinoma (HCC) specimens from the first affiliated Hospital of Guangxi Medical University in 2016 and pathologically diagnosed as HBV positive primary hepatocellular carcinoma (HCC). Cancer and paracancerous tissues were extracted by Trizol method. The expression levels of m RNA and protein of ADAMTS-18 in cancer tissues and adjacent tissues were detected by Q RT-PCR and immunohistochemical staining respectively. The total RNAs of MHCC-97 Hep G2 and HL-7702 cell lines were extracted, and the m RNA expression of ADAMTS-18 gene was detected by Q RT-PCR. The lentivirus mediated gene silencing and overexpression techniques were used, respectively. ADAMTS-18 gene silencing and expression stabilization of MHCC-97Hep G2HL-7702 cell line were constructed. MTT method, cell scratch assay and Transwell chamber invasion assay were used to detect the proliferation, migration and invasion ability of hepatoma cell lines before and after ADAMTS-18 gene silencing and overexpression, respectively. Results the level of ADAMTS-18 m RNA and protein expression in HCC tissues were significantly higher than those in adjacent tissues (P0.05ADAMTS-18m RNA) and in HL-7702MHCC-97Hep SMMC-7721 BEL-7404 and QGY-7703 cell lines.) the stable transformed cell line MHCC-97Hep Hep G2HL-7702.4 was successfully constructed with ADAMTS-18 silencing and overexpression. The results of scar test and Transwell chamber invasion test showed that, After upregulation of ADAMTS-18 expression, the invasion ability of MHCC-97Hep G2 increased significantly, and the invasion ability of MHCC-97H decreased significantly after down-regulation of ADAMTS-18 expression. The results of MTT assay showed that the proliferation ability of Hep G2HL-7702 cell line increased and decreased after up-regulating or down-regulating the expression of MHCC-97Hep G2HL-7702, and the proliferation ability of MHCC-97Hep G2HL-7702 cell line increased and decreased after down-regulating the expression of MHCC-97Hep Hep G2HL-7702. Conclusion the expression of ADAMTS-18 gene in HBV associated HCC tissues is significantly higher than that in adjacent HCC tissues. Upregulating the expression of ADAMTS-18 gene can promote the migration and invasion ability of MHCC-97H in vitro. Down-regulating the expression of ADAMTS-18 gene could inhibit the migration and invasion ability of MHCC-97H in vitro and up-regulate the proliferation of MHCC-97Hep G2 and HL-7702 cell lines. Down-regulation of ADAMTS-18 gene can inhibit the proliferation ability of the above cell lines.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

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