摘要
目的研究P物质(substance P,SP)对体外培养的人低分化胃癌细胞MKN45内游离钙浓度的影响。方法在RPMI1640培养液中培养人胃癌细胞系MKN45,应用钙特异性荧光指示剂Furu-3/AM负载细胞,用ASN-1377642(NK-1受体拮抗剂)、尼卡地平和不同浓度的SP作用于人胃癌细胞MKN45,采用激光扫描共聚焦显微镜观测胃癌细胞内游离钙浓度的变化。结果在Hanks液(含钙离子)中加入10、50及100nmol/L SP可以显著升高胃癌细胞内钙离子浓度(P<0.05),并且呈剂量依赖性(P<0.05);在无外钙时,SP引起细胞内钙离子浓度升高的幅度低于有外钙时(P<0.05);在Hanks液中加入NK-1受体拮抗剂ASN-1377642和钙通道阻滞剂尼卡地平孵育后,SP引起胃癌细胞内钙离子浓度升高的幅度显著降低,与Hanks液组比较差异有统计学意义(P<0.05)。结论SP可以显著升高胃癌细胞内钙离子浓度,细胞内钙离子浓度的升高源于内钙释放和外钙内流。
Objective To study the effect of substance P （SP） on intracellular free calcium concentration in human poorly-differentiated gastric cancer cell in vitro. Methods Human gastric cancer cell line MKN45 was cultured in RPMI 1640. Then the cells were loaded with specific calcium fluorescent probe Furu-3/AM. ASN-1377642 （NK-1 receptor antagonist）, Nicardipine （calcium channel blocker） and different concentrations of SP were used to treat gastric cancer cells. The concentration changes of intracellular free calcium were detected by laser scanning confocal microscope. Results It was found that 10, 50 and 100 nmol/L SP could significantly increase the intracellular free calcium concentration of gastric cancer cells in Hanks solutions, which contain extracellular calcium （P〈 0.05）, and the change was in a dose-dependent manner （P〈0.05）. When there was extracellular calcium existed, the increasing amplitude of intracellular free calcium concentration was significantly higher than that when there was no extracellular calcium （P〈0.05）. And when Hanks solutions were pretreated with ASN-1377642 and Nicardip- ine, the effects of 100 nmol/L SP were partly inhibited, and the concentrations of intracellular free calcium were significantly lower than those in groups without pretreatments （P〈0.05）. Conclusion SP can significantly increase free calcium concentration in the gastric cancer cells. Releasing of stored calcium in the cells and influx of extracellular calcium may contribute to the elevation of intracellular free calcium concentration.
出处
《中国普外基础与临床杂志》
CAS
2008年第1期8-11,共4页
Chinese Journal of Bases and Clinics In General Surgery
基金
全军医药卫生科研基金(编号:01MB123)
陕西省自然科学研究科技基金(编号:2003C209)~~
关键词
P物质
胃癌细胞
钙离子
激光扫描共聚焦显微镜
Substance P Gastric cancer cell Calcium Laser scanning confocal microscope
