白细胞介素增强因子3 对胃癌细胞侵袭转移能力的影响及机制

Effect of ILF3 on metastasis and invasion of gastric cancer cells and mechanism

摘要:

目的 探讨白细胞介素增强因子3（ILF 3）对胃癌细胞侵袭转移能力的影响及相关机制。方法 免疫组织化学技术检测ILF3 在胃癌组织、转移淋巴结组织、正常胃黏膜组织中表达情况；收集患者临床资料，分析ILF3 与胃癌患者临床病理特征的关系。Western blot 检测不同胃癌细胞株中ILF3 表达。合成针对ILF3的小干扰RNA 转染人胃癌细胞株MGC803 ；MTT 法检测MGC803 细胞活性；细胞划痕实验和Transwell小室侵袭实验检测细胞的迁移侵袭能力；实时荧光定量聚合酶链反应（qRT-PCR）和Western blot 检测MMP -7、MMP -9、TIMP -1 基因的mRNA 和蛋白表达情况。结果 胃癌组织中ILF3 蛋白阳性率高于癌旁组织；转移淋巴结中ILF3 蛋白阳性率高于胃癌组织（P <0.05）。胃癌组织中ILF3 蛋白阳性表达与患者的肿瘤浸润深度、分化程度、淋巴结转移、临床分期有关（P <0.05）。各胃癌细胞株中MGC803 的ILF3 蛋白表达最强，抑制MGC803 细胞内源性ILF3 表达后细胞的活性减弱；迁移和侵袭能力也降低（P <0.05）。ILF3-siRNA转染MGC803 后细胞的MMP-2、MMP-7 表达减弱，而TIMP-1 的表达增高（P <0.05）。结论 ILF3 对胃癌的侵袭转移有促进作用，ILF3 可能是通过调节MMP -7、MMP -9、TIMP -1 基因而参与了胃癌的侵袭转移。

Abstract:

Objective To investigate the effect of interleukin enhancer-binding factor 3 (ILF3) on metastasis and invasion in gastric cancer cells and the mechanism. Methods Immunohistochemical technique was applied to test ILF3 expression in gastric cancer tissues, metastatic lymph node, normal gastric mucosa tissues. The clinical data were also recorded. Then the relationships between ILF3 protein and clinical data were explored. Western blot assay was used to test the ILF3 protein level in different gastric cell lines. ILF3-siRNA was synthesized and transfected into MGC803 cell line. MTT assay was applied to detect the cell activity. The ability of MGC803 migration and invasion was observed via wound healing assay and Transwell chamber assay. The mRNA and protein expressions of MMP7,MMP9 and TIMP1 were also detected by qRT-PCR and Western blot. Results The positive rate of ILF3 protein in the gastric cancer tissues was significantly higher than that in the normal gastric mucosa tissues, and the positive rate of ILF3 protein in the metastatic lymph node was higher than that in the gastric cancer tissues (P < 0.05). The expression of ILF3 was related to tumor invasion, differentiation, lymph node metastasis and TNM stage (P < 0.05).In different gastric cell lines, the strongest ILF3 protein expression was found in MGC803 cell line, and the cell activity, migration and invasion ability of MGC803 significantly decreased after ILF3-siRNA was transfected into MGC803 cells (P < 0.05). After transfection, the expressions of MMP7 and MMP9 decreased, whereas the expression of TIMP-1 increased (P < 0.05). Conclusions ILF3 gene could promote migration and invasion of gastric cancer by regulating MMP7, MMP9 and TIMP1 genes.