溶血磷脂酸酰基转移酶β促进人肺成纤维细胞的增殖、迁移与黏附

LPAAT β promotes proliferation,migration and adhesion of HLF cells

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 evaluate the expression of lysophosphatidic acid acyhransferase （LPAAT） 15 in different cancer cell lines and to investigate the effect of LPAAT 15 oyerexpression on cell proliferation, migration and adhesion in vitro. Methods Western blotting assay was used to detect the expression of LPAAT 15. The cDNA of LPAAT 13 was cloned into pcDNA3.1A（ ＋ ） and transfected into human lung fibroblast cells （HLF）. The proliferation of LPAAT 15 stably-ex- pressing HLF cells was analyzed by MTI assay. Wound healing assay was applied to evaluate the cell migration ability and adhesion assay was employed to examine the adhesion of LPAAT 15 stably-expressing HLF ceils to an extracellular matrix, Ma- trigel. Results LPAAT ~ protein expression levels were different in tumor cells. LPAAT 15 stably-expressing HLF cells dis- played elevated cell proliferation and migration ability. Moreover, the transfected cells adhered to Matrigel were also in- creased. Conclusion LPAAT 15 is universally expressed in eukaryotic and tumor cells, and its overexpression in HLF cells promotes proliferation, migration and adhesion ability of cells, indicating the important role of LPAAT 15 in cell growth in vitro.