重组人生长激素联合5-氟尿嘧啶对表达或不表达生长激素受体人肝癌细胞的体外干预

Effects of Recombinant Human Growth Hormone Combined with 5-Fluorouracil on GHR^+ or GHR^- Human Liver Cancer Cell Lines in vitro

目的研究重组人生长激素(rhGH)在体外对表达或不表达生长激素受体(GHR)的人肝癌细胞系的影响。方法采用免疫组织化学方法检测人肝癌细胞系的GHR表达。每种肿瘤细胞系分4组进行处理:未处理组、5-氟尿嘧啶(5-Fu)处理组、rhGH处理组及5-Fu+rhGH联合处理组。采用四甲基偶氮唑蓝(MTT)比色法及流式细胞术分析不同浓度rhGH及其与5-Fu合用对人肝癌细胞系的生长抑制、凋亡、细胞周期及增殖指数(PI)等的影响。结果Bel-7402细胞表达GHR;与未处理组相比,各浓度rhGH组的生长率、G2/M期比例和PI显著升高,细胞凋亡率显著降低(P均<0.05);与5-Fu处理组相比,各浓度rhGH+5-Fu组的G2/M期比例和PI显著升高,抑制率和细胞凋亡率显著降低(P均<0.05)。SMMC7721不表达GHR;不同浓度rhGH对该细胞系的分裂增殖无明显影响(P均>0.05);rhGH+5-Fu联合处理组与5-Fu处理组间的细胞抑制率、凋亡率及PI差异也无显著性(P均>0.05)。结论rhGH在体外能促进GHR+的肝癌细胞增殖,减弱5-Fu对GHR+细胞的抗癌作用,但对GHR-的肝癌细胞无明显影响,也不能明显干扰5-Fu对GHR-细胞的抗癌功能。

Objective To investigate the effects of recombinant human growth hormone （rhGH） on growth hormone receptor （GHR）^＋ or GHR^- human liver cancer cell lines in vitro. Methods The expression of GHR in human liver cancer cell lines was detected by immunohistochemistry. Four groups were assigned according to the cell line treatment modes： untreated group, 5- Fluorouracil （5-Fu） -treated group, rhGH-treated group and rhGH ＋ 5-Fu combination group. By MTT assay and flow cytometry, effects with different concentrations of rhGH and/ or in combination with 5-Fu on the cell inhibitory rate, apoptosis rate, cell cycle, and proliferation index （PI） of human liver cancer cell lines Bel-7402 and SMMC7721 were analyzed. Results GHR was found in Bel- 7402 cells. Compared with the untreated group, the growth rates, percentages of cells in G2/M phase, and PI significantly increased in groups with different concentrations of rhGH, with cell apoptosis rates significantly decreased （ P 〈 0.05 ）. Compared with the 5-Fu group, the percentages of cells in G2/M phase significantly increased and the inhibitory rates and cell apoptosis rates significantly decreased in groups treated with different concentrations of rhGH ＋ 5-Fu. No GHR was expressed in SMMC7721 cell line, and different concentrations of rhGH had no remarkable effect on the division and proliferation of this cell line （ P 〉 0.05 ）. The cell inhibitory rate, apoptosis rate, and PI had no significant differences between 5-Fu-treated group and rhGH ＋ 5-Fu combination group （ all P 〉 0.05 ）. Conclusions rhGH can stimulate the growth of GHR ＋ tumor cell lines such as Bel-7402 and weaken the inhibitory effect of 5-Fu on GH^＋ tumor cells. However, such effect was not remarked for GHR^- tumor cells in vitro.