Lmo4在胃癌发生发展过程中的作用及其机制

发布时间：2018-06-09 22:48

  本文选题：Lmo4 + 胃癌　； 参考：《江苏大学》2017年硕士论文

【摘要】：目的:探讨Lmo4在胃癌和癌旁组织、不同胃癌细胞株中的表达差异,阐明Lmo4在胃癌发生发展中的作用及机制,为胃癌的分子诊断与靶向治疗提供新思路。方法:应用q RT-PCR和免疫组织化学,检测90例胃癌患者的癌组织及对应癌旁组织中Lmo4mRNA的表达差异,分析Lmo4表达与胃癌临床病理因素的相关性。采用Western blot方法比较不同人胃癌细胞株中Lmo4的表达,筛选高表达和低表达细胞。通过RNA干扰技术抑制BGC-823胃癌细胞中Lmo4的表达,应用平板克隆形成实验、Transwell迁移实验、划痕试验、CCK8实验及Western blot方法检测Lmo4被沉默后对胃癌细胞的生长迁移能力及细胞通路(Vimentin通路、MMP9通路、TGF-β通路、AKT通路、β-Catenin通路)的影响。用pcDNA构建过表达Lmo4质粒Lmo4-pcDNA,并通过Western blot检测其对胃癌细胞增殖、迁移、凋亡及EMT相关蛋白的影响,运用细胞周期试验评价Lmo4过表达后增殖情况。以细胞计数、平板克隆形成实验和Transwell迁移实验评价胃癌细胞生长和迁移能力的变化。结果:q RT-PCR结果显示胃癌组织中Lmo4相对表达量明显高于对应的癌旁组织(P0.001),且Lmo4在胃癌组织中的表达与肿瘤的分型有关(P0.001)、浸润程度有关(P0.001),而与年龄、性别、肿瘤大小无关。免疫组化结果发现Lmo4基因特异性表达于胃癌细胞(胞浆与胞核),而在癌旁组织或肠化生组织中未见其表达。高表达Lmo4的BGC-823胃癌细胞迁移能力显著强于低表达Lmo4的MGC-803胃癌细胞。沉默Lmo4后的细胞的迁移能力显著减弱,细胞克隆形成数量减少,促进了细胞凋亡,抑制EMT发生,使E-cadherin表达升高,TWIST、N-cadherin、Vimentin表达降低;Lmo4过表达在体外促进胃癌细胞生长和迁移,抑制了细胞凋亡;促进EMT的发生,使E-cadherin表达降低,TWIST、N-cadherin、Vimentin表达升高。结论:Lmo4在胃癌的发生、进展中具有重要作用,可能通过诱导胃癌细胞发生EMT而促进胃癌的转移。Lmo4不仅可作为胃癌分子诊断一个新的分子标志,而且可能成为治疗胃癌的一个新的靶标。
[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.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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本文编号：2001091