胆管癌细胞增殖与生长激素-胰岛素样生长因子轴的关系

Relationship between the axis of GH-IGFs and the proliferation of bile duct cancer cell line QBC939 in vitro

目的探讨生长激素（GH）在体外对胆管癌细胞QBC939增殖的影响及其可能机制。方法将处于生长对数期的胆管癌细胞QBC939随机分为实验组（GH组）和对照组（Ns组），GH组按剂量和培养时间分为50ug/L 2h（GH50-2h），50ug／L24h（GH50-24h），100ug/L 2h（GH100—2h），100ug／L 24h（GH100—24h）四个亚组，NS组按培养时间也分为NS-2h、NS-24h两个亚组，2h和24h后分别吸取上清用酶联免疫吸附试验法检测胰岛素样生长因子（IGF）1／2并将细胞计数；胆管癌细胞用不同浓度GH培养24h后固定，以流式细胞仪测定细胞周期；同时在不同浓度GH干预的培养液中行细胞爬片后固定，用原位杂交法检测IGF12受体的mRNA（IGF1RmRNA／IGF2RmRNA）。结果培养液中加入GH2h后，QBC939细胞无明显增多（P〉0．05），但24h后细胞数目增多明显，差异有统计学意义（P〈0．05），且24h后流式细胞仪检测显示，与NS组（S％：34．60±1．37；PI：0．42±0．01）比较，GH组S％和细胞增值指数（PI）[GH50组：S％（39．36±1．62；PI：0．48±0．02）；GH100组：（S％：45．74±2．15；PI：0．53±0．02）]也明显增加（P〈0．05）。IGF1RmRNA／IGF2RmRNA在胆管癌中呈阳性表达，且GH可诱导细胞IGF1RmRNA表达，但不诱导IGF2RmRNA的表达增强。结论GH在体外能促进胆管癌QBC939细胞的增殖和分化，其机制可能是通过GH—IGF1-IGF1R轴发挥作用的。

Objective To explore the proliferation and the possible mechanism of human growth hormone on bile duct cancer cell line （ QBC939 cell） in vitro. Methods The QBC939 cells during exponential growth stage were harvested and divided into experimental group （group GH ） and control group （ group NS）. The experimental group was divided into 4 sub-groups （50 ug/L for 2 h,50 ug/L for 24 h, 100 ug/L for 2 h, 100 ug/L for 24 h） according to the doses of GH and culture time. The control group was also divided into two sub-groups （NS-2 h,NS-24 h） according to the culture time. After 2 and 24 h, insulin-like growth factor （IGF） 1/2 was detected by using EIJSA respectively. The QBC939 cells cultured for 24 h with various GH concentrations were made into single cell suspensions and the samples underwent subsequent cell cycle evaluation. IGF1R mRNA/IGF2R mRNA were tested with the method of hybridization in vitro. Results There was no statistical significance of the difference between group GH and group NS after 2 hour＇ s culture （ P 〉 0.05 ）. GH stimulated cell growth in vitro and also elevated percent S phase and proliferation index（PI） of cells after 24h＇ s culture （ P 〈 0.05 ）. IGF1R mRNA and IGF2R mRNA were expressed in QBC939 in contrast to blank control ,and the expression of IGF1R mRNA boosted up according to the dose of GH,but IGF2R mRNA was not. Conclusion CH stimulates QBC939 cells growth and proliferation in vitro and the possible mechanism is acted by the axis of GH-IGF1-IGF1R.