hPPARγ2高表达细胞模型的建立及与细胞恶性程度的关系

Establishment of hPPARγ2-overexpressing cell model and the relationship with tumor malignancy

目的:构建人过氧化物酶体增殖物激活受体γ2(hPPARγ2)的真核表达载体并建立hPPARγ2高表达细胞株,探讨hPPARγ2表达与肿瘤形成及恶性程度的关系。方法:利用常规分子生物学方法构建hPPARγ2真核表达载体,并转染NIH3T3细胞及人乳腺癌MCF-7,MDA-MB-231细胞。利用细胞穿透人工基底膜实验及致瘤性实验检测转染后细胞恶性程度的改变。采用RT-PCR和Western blot的方法检测hPPARγ2以及ERK,p-ERK的表达情况。结果:成功构建hPPARγ2的真核表达载体并建立了hPPARγ2高表达的细胞模型,转染后的细胞侵袭能力增强、致瘤性增加。结论:hPPARγ2的高表达与肿瘤高侵袭、恶性程度的增高呈正相关。

Objective：To establish hPPARγ2 plasmid and cell lines overexpressing hPPARγ2,then to investigate the relationship of hPPARγ2 with tumor malignancy.Methods：By common methods of molecular biology,hPPARγ2 plasmid was established and stably transfected into cells.The malignancy of the transfected cells was detected by invasion through reconstituted basement membrane assay and oncogenicity assay.The levels of hPPARγ2,ERK,and p-ERK in the stably transfected cells were analyzed by RT-PCR and Western blot assays.Results：In the stably transfected cells,hPPARγ2 increased tumor formation and invasion abilities.These changes related to ERK phosphorylation.Conclusion：hPPARγ2 contributes to tumor formation and tumor growth,which is mediated by the ERK signal pathway.