生长激素干预肝癌细胞及共培养脐静脉内皮细胞生长

Effects of recombinant human growth hormone on ECV304 co-cultured with hepatoma cells in vitro

目的观察重组人生长激素（rhGH）对人肝癌细胞株SMMC-7721和Bel-7402，共培养脐静脉内皮细胞株ECV304增殖的影响。方法ECV304及其与Bel-7402、SMMC-7721共培养三组分别行rhGH50及250μg／L、贝伐单抗50mg／L及其联合rhGH250μg/L的干预。Bel-7402和SMMC-7721分别接受两浓度rhGH干预。流式细胞术测细胞周期比。逆转录-聚合酶链反应（RT—PCR）／酶联免疫吸附试验（ELISA）法测定肝癌细胞血管内皮细胞生长因子（VEGF）mRNA／蛋白表达。结果Bel-7402表达生长激素受体（GHR），SMMC-7721不表达GHR；两剂量rhGH干预后Bel-7402的PI指数（42．69±0.40和44．10±0．19）较空白组（39．40±0.48）明显升高，同环境ECV304的PI指数显著升高[（45．62±0．87）％和（47．64±1．28）％，呈浓度依赖（P〈0．05）；贝伐单抗干预后，Bel-7402共培养ECV304的PI指数显著下降[（42．69±1．05）％和（42．84±0．77）％，P〈0．05）。与对照组[mRNA／蛋白：（14．179±3．110）ng／L/0．171±0．064]比较，两rhGH浓度干预，Bel-7402表达VEGFmRNA／蛋白水平也显著增加[（125．499±2．680）和（131．312±2．560）ng／L／0．296±0.024和0．460±0．037，P〈0．05]；SMMC-7721各组均未出现类似情况。结论rhGH明显促GHR阳性表达人肝癌细胞株Bel-7402增殖，促其过量分泌VEGF致共培养内皮细胞ECV304增殖。

Objective To observe the ECV304 proliferation which co-cultured with GHR（ ＋/-） hepatoma ceils by rhGH intervention. Methods Three groups were assigned：ECV304 group, Bel-7402/ ECV304 ,/ECV304FCM co-cultured group. Each group was treated with rhGH 50 μg/L、rhGH 250 μg/L, Bevacizumab 50 mg/L,Bevacizumab 50mg/L/rhGH 250 μg/L. was respective treated with rhGH 50 μg/ L,rhGH 250 μg/L. Cycle ratio were measured. RT-PCR was selected to display the VEGF mRNA expression of hepatoma ceils, and ELISA was selected to display the VEGF concentration. Results Hepatoma cell Bel-7402 expressed GHR,but there was no GHR expressed in Hepatoma cell SMMC-7721. The proliferation indices （PI） of Bel-7402 were significantly increased in rhGH group （42.69± 0.40 and 44.10± 0.19） compared with control group （39.40 ± 0.48 ）, as well as ECV304 cultured with Bel-7402 （45.62 ± 0.87 ） % and （47.64 ± 1.28 ） % , which were showing rhGH dose-dependent manner （ P 〈 0.05 ）. Treased with Bevacizumab, the proliferation indices （PI） of ECV304 cultured with Bel-7402 were significandy decreased [ （42.69 ± 1.05 ） % and （42.84 ± 0.77 ） %, P 〈 0.05 ]. Compared with control group [ （14. 179±3.11 ） ng/L/（0. 171 ± 0. 064）] ,the VEGF mRNA expression and the VEGF secretion of Bel-7402 were significantly higher after rhGH intervention [ （125. 499 ± 2.68 ） and （131. 312 ± 2.56 ） μg/L/0. 296 ± 0. 024 and 0. 460 ± 0. 037, P 〈 0.05 ]. However, we have not seen the manifestation in SMMC-7721GHR（ - ）. Conclusion The rhGH could stimulates the proliferation of GHR（ ＋ ） hepatoma cells, induces the VEGF secretion and promotes the proliferation of ECV304 cultured with Bel-7402.