生长激素对结肠癌细胞侵袭和迁移的影响

The effect of growth hormone on invasion and migration of colon cancer cells

目的 探讨生长激素(GH)对结肠癌细胞增殖、周期及侵袭迁移的影响及其可能的作用机制。方法 在体外培养生长激素型垂体腺瘤GH3细胞,通过ELISA检测GH3细胞培养上清液中GH分泌情况。取对数生长期的人结直肠腺癌LoVo细胞随机分为对照组和实验组,对照组中加入磷酸盐缓冲液(PBS),实验组中加入高浓度重组人GH。两组细胞在相同条件下进行体外培养,通过CCK-8法检测细胞增殖水平,流式细胞仪检测细胞周期,Transwell法检测细胞迁移和侵袭能力,Western blot检测细胞E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和蜗牛蛋白-1(Snail-1)的表达水平。结果 GH3细胞可分泌大量GH,其培养上清液GH浓度为(1 208±9)ng/mL。GH在一定浓度范围内呈剂量依赖性促进细胞生长,GH 200 ng/mL为其最佳干预浓度,用作后续实验浓度。与对照组比较,实验组中细胞周期从G_(1)期向S期和G_(2)转变,G_(1)期细胞比率下降,S期、G_(2)期细胞比率升高(P<0.05);与对照组比较,实验组细胞侵袭和迁移数量增加(P<0.05),细胞N-cadherin、Vimentin和Snail-1表达水平上调,E-cadherin表达水平下调(P<0.05)。结论 高浓度GH可促进结肠癌细胞增殖、侵袭及迁移,诱导细胞周期从G_(1)期向S期和G_(2)转变,其机制可能与高浓度GH促进结肠癌细胞上皮间质转化(EMT)相关。

Objective To explore the effects of growth hormone(GH)on the proliferation,cycle,invasion,and migration of colon cancer cells and its possible mechanism.Methods GH3 cells with growth hormone-type pituitary adenoma were cultured in vitro,and the secretion of growth hormone in the supernatant of GH3 cells was detected by ELISA.Colon cancer LoVo cells in logarithmic growth phase were randomly divided into the control group and the experimental group.PBS was added to the control group,while high concentrations of recombinant GH were added to the experimental group.The two groups of cells were cultured in vitro under the same conditions.CCK-8 method was used to detect the proliferation of the cells.Flow cytometry was used to detect the cell cycle.Transwell assay was used to detect the effect of growth hormone on the invasion and migration of the cells.Western blot was used to detect the expressions levels of E-cadherin,N-cadherin,Vimentin,and Snail-1 proteins in the cells.Results The results of ELISA showed that GH3 cells could secrete a large amount of GH,and the concentration of GH in the supernatant was(1208±9)ng/mL.GH promoted cell growth in a dose-dependent manner within a certain concentration range,and GH 200 ng/mL was the optimal intervention concentration for subsequent experiments.Compared with the control group,the cell cycle in the experimental group changed from G_(1) phase to S phase and G_(2) phase,the ratio of G_(1) phase cells decreased,and the ratio of S phase cells and G_(2) phase cells increased(P<0.05).Compared with the control group,the number of the cell invasion and migration increased in the experimental group(P<0.05),the expression levels of N-cadherin,Vimentin,and Snail-1 was up-regulated,while the expression level of E-cadherin was down-regulated(P<0.05).Conclusion High concentration of GH promotes the proliferation,invasion and migration of colon cancer cells,and induces the transition of cell cycle from G_(1) to S and G_(2) phases.The mechanism may be related to the epithelial-mesenchymal transition(EMT)of colon cancer cells promoted by high concentration of GH.