南方医科大学学报 ›› 2017, Vol. 37 ›› Issue (06): 774-.

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血清限制对人滋养细胞株HTR-8/SVneo细胞胰岛素样生长因子-1表达及细胞侵袭力的影响

周尚谦,沈朗,李雪媛,谢晓珍,芮塬,陈年坤,王志坚   

  • 出版日期:2017-06-20 发布日期:2017-06-20

Effect of serum restriction on insulin like growth factor-1 expressions and invasiveness in human trophoblast HTR-8/SVneo cells in vitro

  • Online:2017-06-20 Published:2017-06-20

摘要: 目的探讨血清限制对人滋养细胞株HTR-8/SVneo细胞胰岛素样生长因子-1(IGF-1)、基质金属蛋白酶-2(MMP-2)表达及 细胞侵袭力的影响。方法分别以1%、5%及10%胎牛血清(FBS)处理HTR-8/SVneo 细胞48 h 后,荧光定量PCR检测细胞的 IGF-1及MMP-2 mRNA水平,细胞免疫荧光检测细胞IGF-1、MMP-2蛋白表达,MTT检测细胞增殖能力,Transwell小室实验检 测细胞侵袭力;进一步添加人重组IGF-1处理后,观察HTR-8/SVneo细胞MMP-2表达、细胞增殖及侵袭能力的改变。结果随 着血清浓度降低,细胞增殖能力明显下降(P<0.05);血清限制抑制细胞IGF-1、MMP-2的mRNA及蛋白表达(P<0.05),导致细胞 侵袭力下降(P<0.05);补充人重组IGF-1能显著上调HTR-8/SVneo细胞MMP-2 表达水平(P<0.05),促进HTR-8/SVneo细胞侵 袭(P<0.05)。结论血清限制下调人滋养细胞株HTR-8/SVneo细胞IGF-1表达,可能通过引起其MMP-2表达降低来介导细胞 侵袭力下降;补充人重组IGF-1有助于促进HTR-8/SVneo细胞增殖及侵袭。

Abstract: Objective To explore the effect of serum restriction on the invasiveness and expressions of insulin-like growth factor-1 (IGF-1) and matrix metalloproteinase-2 (MMP-2) in human trophoblast HTR-8/SVneo cells in vitro. Methods HTR-8/ SVneo cells were cultured in the presence of 1%, 5%, or 10% fetal bovine serum (FBS) for 48 h. Fluorescence quantitative PCR and immunofluorescence staining were employed to examine the changes in IGF-1 and MMP-2 expressions at both the mRNA and protein levels in HTR-8/SVneo cells; MTT assay and Transwell invasion assay were used to assess the changes of the cell proliferation and the cell invasion ability, respectively. MMP-2 expression, cell proliferation and invasiveness were also assessed in the cells treated with recombinant human IGF-1. Results HTR-8/SVneo cells exhibited significantly lowered cell proliferation in cultures containing low concentrations of FBS (P<0.05). The expressions of IGF-1 and MMP-2 at both mRNA and protein levels were significantly down-regulated and the invasiveness was significantly lowered in cells cultured in the medium containing 1% FBS as compared with those of cells cultured in the presence of 5% and 10% FBS (P<0.05). Treatment of the cells with recombinant human IGF-1 significantly up-regulated MMP-2 expression (P<0.05) and increased the cell invasiveness (P<0.05). Conclusions FBS restriction down-regulates IGF-1 expression in human trophoblast HTR-8/SVneo cells and suppress the cell invasiveness possibly by suppressing MMP-2 expression. Treatment with recombinant human IGF-1 can up-regulate MMP-2 expression and promote the invasiveness of HTR-8/SVneo cells.