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目的分析溶血磷脂酸酰基转移酶β(lysophosphatidic acid acyltransferase,LPAATβ)在多种细胞系中的表达情况及其过表达对细胞体外增殖、迁移与黏附的影响。方法通过免疫印迹实验检测LPAATβ在多种细胞中的表达水平;构建pcDNA3.1A(+)/LPAATβ真核表达载体,转染人肺成纤维细胞(human lung fibroblast,HLF)并筛选稳定转染细胞株;通过MTT细胞增殖实验分析过表达LPAATβ对HLF细胞体外增殖的作用,通过划痕修复实验分析稳定株细胞的体外迁移能力,利用细胞黏附实验分析稳定株细胞对细胞外基质成分Matrigel的黏附情况。结果 LPAATβ蛋白在肿瘤细胞中表达水平存在差异。体外生物学实验显示,稳定转染LPAATβ的HLF细胞增殖能力明显增强,细胞迁移能力显著提高,与细胞外基质成分Matrigel的黏附能力显著增强。结论 LPAAT家族成员LPAATβ在人类多种细胞中的表达具有普遍性,其过表达明显促进HLF细胞的增殖、迁移及黏附能力,提示该基因在细胞体外生长过程中具有重要作用。
Objective To analyze the expression of lysophosphatidic acid acyltransferase (LPAATβ) in various cell lines and its effect on cell proliferation, migration and adhesion in vitro. Methods The expression of LPAATβ in various cells was detected by Western blotting. The eukaryotic expression vector pcDNA3.1A (+) / LPAATβ was constructed and transfected into human lung fibroblasts (HLF) The effect of LPAATβ overexpression on the proliferation of HLF cells was analyzed by MTT cell proliferation assay. The migration ability of stable cells in vitro was analyzed by scratch repair assay. The adhesion of stable cells to Matrigel was analyzed by cell adhesion assay . Results LPAATβ protein expression levels in tumor cells there are differences. In vitro biological experiments showed that the proliferation of HLF cells stably transfected with LPAATβ was significantly enhanced, the ability of cell migration was significantly increased, and the adhesion ability to Matrigel was significantly enhanced. Conclusion LPAAT family member LPAATβ is universally expressed in many human cells. Overexpression of LPAATβ can significantly promote the proliferation, migration and adhesion of HLF cells, suggesting that LPAATβ plays an important role in the in vitro growth of HL-60 cells.