Notch1/Hes1信号通路在调控低分化胃腺癌细胞钙离子和侵袭能力中的作用

目的探讨Notch1/Hes1信号通路对胃癌细胞钙离子水平的影响及其与侵袭的关系。方法采用免疫组化法检测32例原发性低分化胃腺癌组织及32例癌旁正常组织中Notch1、Hes1、Calpain-1和E-cadherin蛋白的表达。将MGC-803细胞分为对照组、Lipofectamine 2000组、Notch1 siRNA组和Hes1 siRNA组,采用Transwell法对比分析Notch1和Hes1基因沉默MGC-803细胞侵袭能力。应用Flou-4实验检测细胞钙离子水平。运用Western blot法检测Notch1、Hes1、Calpain-1和E-cadherin蛋白的表达。结果 32例低分化胃腺癌组织中Notch1、Hes1、Calpain-1表达(93.75%、96.88%、96.88%)明显高于癌旁正常组织(21.88%、18.75%、25.0%)(P<0.05),E-cadherin表达(25.0%)明显低于癌旁正常组织(93.75%,P<0.05)。MGC-803细胞Transwell和Flou-4实验可见,Notch1 siRNA组和Hes1 siRNA组癌细胞的迁移量明显低于对照组(P<0.05),癌细胞钙离子水平也明显低于对照组(P<0.05)。Western blot检测显示,Notch1 siRNA组和Hes1 siRNA组癌细胞Calpain-1和vimentin的相对含量低于对照组(P<0.05),而E-cadherin相对含量高于对照组(P<0.05)。结论低分化胃腺癌细胞侵袭能力与Notch1/Hes1蛋白及细胞钙离子有关,Notch1/Hes1信号通路参与Calpain-1相关的细胞钙离子调控。

Notch1/Hes1 signaling pathway associated with regulation of calcium ions and invasion in poorly differentiated gastric adenocarcinoma

Purpose To investigate the effect of Notch1/Hes1 signaling pathway on the calcium ions and invasion in gastric adenocarcinoma.Methods Immunohistochemistry was used to detect the expression of Notch1,Hes1,E-cadherin,and Calpain-1 in 32 cases of primary poorly differentiated gastric adenocarcinoma and para-cancerous tissues.MGC-803 cells were divided into control group,Lipofectamine 2000 group,Notch siRNA group and Hes1 siRNA group.Transwell was used to detect and analysis the invasive ability of Notch1 and Hes1 gene silence MGC-803 cells.Flou-4 was used to detect the level of calcium ions in different groups of cells.The expressions of Notch1,Hes1,Calpain-1 and E-cadherin in MGC-803 cells was detected by Western blot.Results The expression of Notch1,Hes1,and Calpain-1 in 32 poorly differentiated gastric adenocarcinoma(93.75%,96.88%and 96.88%,respectively)was significantly higher than that in paracancerous tissues(21.88%,18.75%and 25.0%,respectively)by immunohistochemistry(P<0.05),and meanwhile,the expression of E-cadherin(25.0%)was significantly decreased compared with paracancerous tissues(93.75%,P<0.05).Transwell and Flou-4 experiments of MGC-803 cells showed that the migration of cancer cells in Notch1 siRNA group and Hes1 siRNA group was significantly decreased compared with the control group(P<0.05),and the calcium ion level of cancer cells was also significantly lower than that in the control group(P<0.05).Western blot showed that the relative content of Calpain-1 and vimentin in Notch1 siRNA group and Hes1 siRNA group was significantly lower than that in control group(P<0.05),while the relative content of E-cadherin in Notch1 siRNA group and Hes1 siRNA group was significantly higher than that in control group.Conclusion The invasion in poorly differentiated gastric adenocarcinoma is associated with Notch1/Hes1 and calcium ions,in which Notch1/Hes1 signaling pathway involved in the regulation of calcium ions via Calpain-1.