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投稿时间：2022-04-14 网络发布日期：2022-11-20
投稿时间：2022-04-14 网络发布日期：2022-11-20
中文摘要: 目的 探讨LIM结构域蛋白4（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。方法应用qRT-PCR和免疫组织化学技术，检测2019年4月至2020年6月浦口区中心医院90例胃癌患者的癌组织及对应癌旁组织中Lmo4 mRNA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、CCK-8实验及western blot等方法检测Lmo4沉默对胃癌细胞生长迁移能力及细胞通路的影响。构建Lmo4过表达质粒Lmo4-pcDNA，并通过western blot检测其对胃癌细胞增殖、迁移及上皮细胞间质转化（EMT）相关蛋白的影响，运用细胞周期试验评价Lmo4过表达后细胞增殖情况的改变。结果 qRT-PCR和免疫组织化学结果均显示胃癌组织中Lmo4相对表达量明显高于对应的癌旁组织（P<0.01），且Lmo4在胃癌组织中的表达与肿瘤的分型、浸润程度有关（P<0.01），而与年龄、性别、肿瘤大小无关（P>0.05）。沉默Lmo4后的胃癌细胞迁移能力显著减弱，细胞克隆形成数量减少，抑制EMT发生；Lmo4过表达促进胃癌细胞生长和迁移，促进EMT的发生，调节TGF-β信号通路（P<0.05）。结论 Lmo4在胃癌的发生及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。
Abstract:Objective To investigate the expression difference of LIM domain protein 4 (Lmo4) in gastric cancer, adjacent tissues and different gastric cancer cell lines, and to clarify the role and mechanism of Lmo4 in the genesis and development of gastric cancer. Methods The expression difference of Lmo4 mRNA in 90 gastric cancer tissues and the corresponding paracancerous tissues collected at Pukou District Central Hospital from April 2019 to June 2020 were detected by qRT-PCR and immunohistochemistry. The correlation between Lmo4 expression and clinicopathological factors was analyzed. Western blot was used to compare the expression of Lmo4 in different human gastric cancer cell lines for screening high and low expression cells. The expression of Lmo4 in BGC-823 gastric cancer cells was inhibited by RNA interference technique. Clone formation assay, Transwell migration test, wound-healing assay, cell counting kit-8 and western blot were performed to explore the impact of silencing Lmo4 on gastric cancer cell proliferation and metastasis ability and cell pathway. Lmo4 overexpression plasmid Lmo4-pcDNA was constructed, and its effects on the proliferation, migration and EMT related proteins of gastric cancer cells were detected by western blot. Cell cycle test was used to evaluate the changes of cell proliferation after overexpression of Lmo4. Results The expression of Lmo4 in gastric cancer tissues was significantly higher than that in paracancerous tissues (P<0.01), and the expression of Lmo4 in gastric carcinoma was related to tumor type and infiltration degree (P<0.01), but not to age, gender, tumor size (P>0.05). After silencing Lmo4, the migration ability of gastric cancer cells and the number of cell clones was reduced, and the occurrence of epithelial mesenchymal transformation (EMT) was inhibited. Overexpression of Lmo4 promoted the growth and migration of gastric cancer cells, promoted the occurrence of EMT and regulates TGF-β signaling.Conclusions Lmo4 plays an important role in the development and progression of GC and may promote the metastasis of GC by inducing epithelial-mesenchymal transition in GC cells.
keywords: LIM domain protein 4 Gastric cancer Metastasis Epithelial-mesenchymal transition TGF-β signaling pathway
文章编号： 中图分类号： 文献标志码：A
基金项目:南京医科大学科技发展基金（NMUB2020335）
| 作者 | 单位 |
| 梁怀盼1 | 1. 江苏省人民医院浦口分院 浦口区中心医院检验科，江苏 南京 211800； |
| 许珈豪2 | 2. 江苏大学医学院，江苏 镇江 212000； |
| 陈璐1 | 1. 江苏省人民医院浦口分院 浦口区中心医院检验科，江苏 南京 211800 |
| Author Name | Affiliation |
| LIANG Huai-pan*, XU Jia-hao, CHEN Lu | Department of Laboratory, Pukou Branch of Jiangsu Peoples Hospital, Pukou District Central Hospital, Nanjing, Jiangsu 211800, China |
引用文本：
梁怀盼,许珈豪,陈璐.LIM结构域蛋白4对胃癌细胞转移及增殖能力的作用及机制[J].中国临床研究,2022,35(11):1510-1515.
梁怀盼,许珈豪,陈璐.LIM结构域蛋白4对胃癌细胞转移及增殖能力的作用及机制[J].中国临床研究,2022,35(11):1510-1515.