生长激素释放肽对人胃癌 AGS 细胞的增殖作用。

The proliferative effects of ghrelin on human gastric cancer AGS cells.

机构信息

Department of Gastroenterology, Jinshan Hospital, Fudan University, Shanghai Medical College of Fudan University, Shanghai, China.

出版信息

J Dig Dis. 2012 Sep;13(9):453-8. doi: 10.1111/j.1751-2980.2012.00616.x.

DOI:10.1111/j.1751-2980.2012.00616.x

PMID:22908970

Abstract

OBJECTIVE

To investigate the role of ghrelin in the gastric cancer cell line AGS and its probable mechanism.

METHODS

Cell proliferation was detected by MTT assay after treated with ghrelin or des-acyl ghrelin. The expression of growth hormone secretagogue receptor 1a (GHS-R1a) and 1b (GHS-R1b) mRNA was detected using reverse transcription polymerase chain reaction (RT-PCR). The activity of extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt was measured by Western blot in cells either treated with ghrelin or inhibitors for ERK1/2 and phosphoinositide-3 kinase (PI3K). The distribution of cell cycle phases was determined by flow cytometry analysis of DNA content.

RESULTS

GHS-R1a and GHS-R1b mRNA were expressed in the AGS cells. Ghrelin and des-acyl ghrelin induced AGS cell proliferation at concentrations of 1 nmol/L and 10 nmol/L but had no proliferative effect at a concentration of 100 nmol/L. The treatment of AGS cells with 10 nmol/L of ghrelin and des-acyl ghrelin resulted in the progression of the increased cells in the S phase. ERK1/2 and Akt were activated by ghrelin and des-acyl ghrelin. Specific ERK1/2 inhibitor PD98059 and PI3K inhibitor wortmannin reduced phosphorylation of ERK1/2 and Akt, respectively and blocked ghrelin- and des-acyl ghrelin-induced AGS cell proliferation.

CONCLUSION

Ghrelin and des-acyl ghrelin stimulate the proliferation of gastric cancer cells via the activation of the ERK1/2 and PI3K/Akt pathway.

摘要

目的

研究 ghrelin 在胃癌细胞系 AGS 中的作用及其可能的机制。

方法

用 MTT 法检测 ghrelin 或去酰化 ghrelin 处理后细胞的增殖情况。采用逆转录聚合酶链反应（RT-PCR）检测生长激素促分泌素受体 1a（GHS-R1a）和 1b（GHS-R1b）mRNA 的表达。用 Western blot 法检测 ghrelin 或 ERK1/2 和磷酯酰肌醇-3-激酶（PI3K）抑制剂处理细胞后细胞外信号调节激酶 1/2（ERK1/2）和 Akt 的活性。通过流式细胞术分析 DNA 含量来确定细胞周期各时相的分布。

结果

AGS 细胞中表达 GHS-R1a 和 GHS-R1b mRNA。ghrelin 和去酰化 ghrelin 在 1 nmol/L 和 10 nmol/L 浓度下诱导 AGS 细胞增殖，但在 100 nmol/L 浓度下无增殖作用。10 nmol/L 的 ghrelin 和去酰化 ghrelin 处理 AGS 细胞导致 S 期细胞增多。ghrelin 和去酰化 ghrelin 激活 ERK1/2 和 Akt。ERK1/2 特异性抑制剂 PD98059 和 PI3K 抑制剂 wortmannin 分别降低 ERK1/2 和 Akt 的磷酸化，阻断 ghrelin 和去酰化 ghrelin 诱导的 AGS 细胞增殖。