斯诺普利抑制循环肿瘤细胞粘附于脐静脉内皮细胞的作用研究

Effect of S-nitrosocaptopril on adhesion of circulating tumor cells to umbilical vein endothelial cells

目的:探索斯诺普利(Cap-NO)干预循环肿瘤细胞(CTCs)粘附于人脐静脉内皮细胞(HUVECs)的作用。方法:UV-Vis检测在不同时间点及不同浓度梯度下Cap-NO的特征吸收峰(300 nm)峰值;MTT法检测Cap-NO对HUVECs和HT-29的毒性;粘附实验检测Cap-NO抑制肿瘤细胞HT-29粘附到HUVECs的效率;流式检测Cap-NO对肿瘤细胞粘附分子及整合素活性的作用;qRT-PCR和三磷酸腺苷(ATP)检测Cap-NO对肿瘤细胞HK1、HK2、MMP2的mRNA表达和ATP水平。结果:Cap-NO的特征峰(300 nm)峰值,随着浓度的减小和时间的延长而减小;Cap-NO对2种细胞都表现为低毒性;Cap-NO可抑制肿瘤细胞粘附到人脐静脉内皮细胞,且与给药浓度成正比;对肿瘤细胞的分子CD44具有抑制作用,且与给药浓度成正比;Cap-NO也可抑制HK1、HK2、MMP2的mRNA表达并降低ATP的形成,抑制肿瘤能量代谢。结论:Cap-NO是一种有效的NO供体化合物,在低细胞毒性浓度范围内能有效干预肿瘤细胞粘附于血管内皮细胞,抑制肿瘤细胞能量代谢,可能具有预防肿瘤转移的作用。

Objective:To explore the effect of Cap-NO on the adhesion of circulating tumor cells(CTCs) to human umbilical vein endothelial cells(HUVECs).Methods:UV-Vis was used to detect the characteristic absorption peak(300 nm)of Cap-NO at different time points and different concentration gradients.The cytotoxicity of Cap-NO to HUVECs and HT-29 cells was detected by MTT assay.Adhesion assay was used to detect the Cap-NO adhesion inhibition efficiency of HT-29 tumor cells to HUVECs.Flow cytometric analysis was used to detect the effect of Cap-NO on tumor cells adhesion molecules and integrin activity.q RT-PCR and ATP experiment were used to detect the expression of tumor cells mRNA of HK1,HK2 and MMP2 and adenosine triphosphate(ATP) levels.Results:The characteristic peak of Cap-NO(300 nm)decreased with the decrease of concentration and the prolongation time.Cap-NO showed low toxicity of HT-29 cells and HUVECs.Cap-NO inhibited the adhesion of tumor cells to HUVECs with the concentration of it.Cap-NO inhibited the growth of CD44,which was proportional to the concentration of drug.Cap-NO inhibited the mRNA expression of HK1,HK2 and MMP2 and reduced the formation of ATP to inhibit tumor energy metabolism.Conclusion:The present studies proved that Cap-NO is an effective NO donor compound.It can effectively interfere with the adherence of tumor cells to vascular endothelial cells and inhibit the energy metabolism of tumor cells in a low cytotoxic concentration range.It may have the effect of preventing metastasis of cancer.