本文選題：Lmo4 + 胃癌��； 參考：《江蘇大學(xué)》2017年碩士論文

【摘要】：目的:探討Lmo4在胃癌和癌旁組織、不同胃癌細胞株中的表達差異,闡明Lmo4在胃癌發(fā)生發(fā)展中的作用及機制,為胃癌的分子診斷與靶向治療提供新思路。方法:應(yīng)用q RT-PCR和免疫組織化學(xué),檢測90例胃癌患者的癌組織及對應(yīng)癌旁組織中Lmo4mRNA的表達差異,分析Lmo4表達與胃癌臨床病理因素的相關(guān)性。采用Western blot方法比較不同人胃癌細胞株中Lmo4的表達,篩選高表達和低表達細胞。通過RNA干擾技術(shù)抑制BGC-823胃癌細胞中Lmo4的表達,應(yīng)用平板克隆形成實驗、Transwell遷移實驗、劃痕試驗、CCK8實驗及Western blot方法檢測Lmo4被沉默后對胃癌細胞的生長遷移能力及細胞通路(Vimentin通路、MMP9通路、TGF-β通路、AKT通路、β-Catenin通路)的影響。用pcDNA構(gòu)建過表達Lmo4質(zhì)粒Lmo4-pcDNA,并通過Western blot檢測其對胃癌細胞增殖、遷移、凋亡及EMT相關(guān)蛋白的影響,運用細胞周期試驗評價Lmo4過表達后增殖情況。以細胞計數(shù)、平板克隆形成實驗和Transwell遷移實驗評價胃癌細胞生長和遷移能力的變化。結(jié)果:q RT-PCR結(jié)果顯示胃癌組織中Lmo4相對表達量明顯高于對應(yīng)的癌旁組織(P0.001),且Lmo4在胃癌組織中的表達與腫瘤的分型有關(guān)(P0.001)、浸潤程度有關(guān)(P0.001),而與年齡、性別、腫瘤大小無關(guān)。免疫組化結(jié)果發(fā)現(xiàn)Lmo4基因特異性表達于胃癌細胞(胞漿與胞核),而在癌旁組織或腸化生組織中未見其表達。高表達Lmo4的BGC-823胃癌細胞遷移能力顯著強于低表達Lmo4的MGC-803胃癌細胞。沉默Lmo4后的細胞的遷移能力顯著減弱,細胞克隆形成數(shù)量減少,促進了細胞凋亡,抑制EMT發(fā)生,使E-cadherin表達升高,TWIST、N-cadherin、Vimentin表達降低;Lmo4過表達在體外促進胃癌細胞生長和遷移,抑制了細胞凋亡;促進EMT的發(fā)生,使E-cadherin表達降低,TWIST、N-cadherin、Vimentin表達升高。結(jié)論:Lmo4在胃癌的發(fā)生、進展中具有重要作用,可能通過誘導(dǎo)胃癌細胞發(fā)生EMT而促進胃癌的轉(zhuǎn)移。Lmo4不僅可作為胃癌分子診斷一個新的分子標志,而且可能成為治療胃癌的一個新的靶標。
[Abstract]:Objective: to investigate the difference of Lmo4 expression in gastric cancer and adjacent tissues, and to elucidate the role and mechanism of Lmo4 in the development of gastric cancer, and to provide a new idea for molecular diagnosis and targeted therapy of gastric cancer. Methods: the expression of Lmo4 mRNA in cancer tissues and adjacent tissues of 90 patients with gastric cancer was detected by Q RT-PCR and immunohistochemistry, and the correlation between Lmo4 expression and clinicopathological factors of gastric cancer was analyzed. The expression of Lmo4 in different human gastric cancer cell lines was compared by Western blot, and high and low expression cells were screened. The expression of Lmo4 in BGC-823 gastric cancer cells was inhibited by RNA interference technique. The effect of Lmo4 on the growth and migration of gastric cancer cells after being silenced and the effect of Vimentin pathway, MMP9 pathway, TGF- 尾 pathway and 尾 -Catenin pathway on the growth and migration of gastric cancer cells were detected by scratch test, CCK8 test and Western blot method. Overexpression of Lmo4 plasmid Lmo4-pcDNAwas constructed with pcDNA. the effects of Lmo4-pcDNAs on proliferation, migration, apoptosis and EMT related proteins of gastric cancer cells were detected by Western blot. Cell cycle test was used to evaluate the proliferation of Lmo4 after overexpression. Cell count, plate clone formation assay and Transwell migration assay were used to evaluate the growth and migration ability of gastric cancer cells. Results the relative expression of Lmo4 in gastric cancer tissues was significantly higher than that in adjacent tissues of gastric cancer. The expression of Lmo4 in gastric cancer tissues was related to the type of tumor, and the degree of invasion was related to the degree of invasion, but not to age, sex and tumor size. Immunohistochemical results showed that Lmo4 gene was specifically expressed in gastric cancer cells (cytoplasm and nucleus) but not in paracancerous tissues or intestinal metaplasia. The migration ability of BGC-823 cells with high expression of Lmo4 was significantly stronger than that of MGC-803 cells with low expression of Lmo4. After silencing Lmo4, the migration ability of the cells decreased significantly, the number of cell clone formation decreased, which promoted apoptosis, inhibited the occurrence of EMT, increased the expression of E-cadherin and decreased the expression of Vimentin in TWISTT-N-cadherinine and promoted the growth and migration of gastric cancer cells in vitro. The expression of E-cadherin was decreased and the expression of Vimentin was increased. Conclusion.Lmo4 plays an important role in the carcinogenesis and progression of gastric cancer. It may promote the metastasis of gastric cancer by inducing EMT in gastric cancer cells. Lmo4 can not only be used as a new molecular marker for the molecular diagnosis of gastric cancer. And may become a new target for the treatment of gastric cancer.
【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.2

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相關(guān)期刊論文 前3條

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本文編號：2001091