PDGF-D对人肝癌细胞增殖及VEGF表达影响研究*

The Influence of Platelet-derived Growth Factor D on the Proliferation of Human Hepatocarcinoma Cells and Expression of VEGF

    摘要
  • 摘要: 目的：研究血小板源性生长因子D（PDGF-D）对人肝癌细胞株BEL-7402增殖及其血管内皮生长因子（VEGF）表达的影响。方法：体外培养肝癌细胞株BEL-7402和肝癌旁非瘤性细胞株QSG-7701，采用RT-PCR 方法检测PDGF-D 与PDGFR βmRNA 在BEL-7402和QSG-7701的表达情况；将浓度分别为0（对照）、5、10、20、50、100、200 μ g/mL 的人重组PDGF-DD蛋白加入BEL-7402中，采用四甲基偶氮唑蓝比色法检测肝癌细胞的生长曲线；流式细胞仪检测细胞周期变化；半定量RT-PCR 检测VEGF及PDGFR β mRNA 表达情况，ELISA 检测PDGF-DD干预细胞后培养上清中VEGF蛋白的表达情况。结果：PDGF-D 及PDGFR βmRNA 在BEL-7402中高表达，与QSG-7701相比差异有统计学意义（P<0.05）。 PDGF-DD干预细胞后，可促进人BEL-7402增殖，浓度为100 μ g/mL 时达最高峰；细胞周期分布变化，G0/G1 期细胞数减少，S 期细胞数增加，与对照组相比差异有统计学意义；RT-PCR 结果显示，VEGF 及PDGFR β RI 值，实验组（除5 μ g/mL 组外）与对照组相比差异有统计学意义；ELISA 结果显示，加入PDGF-DD各浓度组VEGF蛋白浓度较对照组增高，差异有统计学意义，并呈量效依赖性关系。结论：PDGF-DD能促进BEL-7402的增殖，上调PDGFR β 及VEGF的表达。PDGF-D 及其信号传导系统在肝癌的发生、发展中可能发挥重要的作用，可作为肝癌预后预测指标和治疗靶点。

     

    Abstract: Objective: To investigate the influence of platelet-derived growth factor D (PDGFD) on the proliferation of BEL-7402 human liver tumor cells and the expression of vascular endothelial growth factor (VEGF). Methods:Human liver tumor cell strain BEL- 7402 and paracarcinoma cell strain QSG-7701 were culturedin vitro . PDGF-D and PDGFR β mRNA expression were assessed by reverse transcription-polymerase chain reaction (RT-PCR). BEL-7402was treated with differ -ent concentrations ( 0, 5, 10, 20, 50, 100 , and 200 μ g/mL) of exogenous PDGF-D. Cel l prol i feration was measured using the MTT assay. The effect of exogenous PDGF-D on the cell cycle of BEL- 7402cells was assessed by flow cytometry. The expression of VEGF was detected by RT-PCR and ELISA in BEL- 7402 cells that were exposed to different concentrations of PDGF-D. Results: PDGF-D and PDGFR β mRNA expression in human l iver tumor cel l strain BEL-7402 was considered significantly higher than that in paracarcinoma cell strain QSG-7701 (P<0.05). Exogenous PDGF-D promoted proliferation of tumor cells in a dose-dependent manner from 10μ g/mL to 100 μ g/mL (P<0.05). The effect of PDGF-D at 100 µg/mL was stronger than at other concentrations. Incubation of tumor cells with PDGF-D markedly decreased the percentage of cells in G0/G 1-phase and increased the percentage of cel ls in S-phase. Compared wi th the control group, VEGF and PDGFR β mRNA expression in the experimental groups (PDGF-D from10µg/mL to 200 µg/mL) was increased ( P<0.05). VEGF pro -tein expression in the experimental groups was significantly enhanced by exogenous PDGF-D in a dose-dependent man -ner, except in the 5 µg/mL group. Similarly, VEGF protein expression in the experimental groups was significantly higher than that in the control group. Conclusion:PDGF-D can promote proliferation of hepatocarcinoma BEL-7402 cells and in-crease expression of VEGF in a dose-dependent manner. PDGF-D may play an important role in the development of HCC and can be used as an index for prognosis evaluation and a target of treatment for liver cancer.

     

    • HTML全文
    • 参考文献(0)
    • 相关文章
    • 施引文献
  • 资源附件(0)

/

返回文章
返回