胃泌素受体拮抗剂与环氧合酶-2抑制剂对胃癌细胞增殖和凋亡的影响  

Effects of gastrin receptor antagonist and cyclooxygenase-2 inhibitor on proliferation and apoptosis of gastric cancer cell

作　　者：孙为豪[1] 苏菡[2] 章礼久[3] 邵耘[1] 许海尘[1] 张涛[1] 薛绮萍[1] 丁国宪[1] 程蕴琳[1] 

机构地区：[1]南京医科大学第一附属医院老年医学科,210029 [2]江苏省省级机关医院内科 [3]安徽医科大学第一附属医院消化科

出　　处：《中华医学杂志》2006年第4期250-254,共5页National Medical Journal of China

基　　金：江苏省卫生厅"135工程"重点人才基金资助项目(苏卫科教[2003]19号)

摘　　要：目的探讨胃泌素受体拮抗剂丙谷胺与特异性环氧合酶-2(COX-2)抑制剂NS-398对人胃腺癌细胞株MKN-45增殖、凋亡的调控作用。方法MKN-45细胞常规培养于RPMI-1640培养液中,细胞长至亚单层后加丙谷胺(终浓度5.0mmol/L)和/或NS-398(10.0μmol/L),连续培养48h。四甲基偶氮唑蓝(MTT)比色分析检查细胞增殖,流式细胞仪检测细胞凋亡,逆转录聚合酶链反应(RT-PCR)和Western印迹法检测凋亡抑制基因bcl-2mRNA及蛋白表达。结果丙谷胺和NS-398协同抑制MKN-45细胞增殖;丙谷胺组、NS-398组和联合用药组的细胞凋亡率分别为24.7%±3.2%,26.7%±3.4%和36.1%±4.6%,显著高于对照组的1.6%±0.6%(均P<0.01);联合用药组的细胞凋亡率明显高于单一用药组(P<0.05)。丙谷胺和NS-398显著下调bcl-2mRNA及蛋白表达(均P<0.05)。结论丙谷胺、NS-398抑制MKN-45细胞增殖,通过下调bcl-2基因表达而诱导细胞凋亡,两者联合具有协同抗癌作用。Objective To investigate the regulative roles of the gastrin receptor antagonist proglumide and the specific cyclooxygenase （COX）-2 inhibitor NS-398 on the proliferation and apoptosis of gastric cancer cells. Methods Human gastric cancer cells of the line MKN-45 were routinely cultured in RPMI-1640 medium supplemented with 10% beat-inactivated fetal calf serum. Subconfluent cell cultures were treated with proglumide at a final concentration of 5 mmoL/L, NS-398 at a final concentration of 10.0 μmol/L, or proglumide in combination with NS-398 for 48 h. The growth and proliferation of MKN-45 cells were analyzed with MTr assay. Flow cytometric analysis was used to detect the apoptosis of the gastric cancer cells. RT-PCR and Western blotting were used to detect the expression of apeptosis-inhibited gene bcl-2 mRNA and protein. Results The apoptosis rates of the cells treated by proglumide, NS-398, and combination of two agents were 24.7% ± 3.2%, 26.7% ± 3.4%, and 36.1% ± 4.6% respectively, all significantly higher than that in the control group （1.6% ± 0.6%, all P〈0.01）. The apeptosis rates of the MKN-45 cells treated with proglumide combined with NS-398 was significantly greater than those of the cells treated by the two agents alone （ both P 〈 0.05 ）. Treatment with proglumide and NS-398 significantly reduced the bcl-2 mRNA and protein expression in the MKN-45 cells （P 〈 0.05 ）. Conclusion Both proglumide and NS-398 inhibit the proliferation and induce the apoptosis of human gastric cells. This apeptosis may be mediated by down-regulation of the expression of apoptosis-inhihited gene bcl-2. Cotreatment with proglumide and NS-398 have synergistic anticancer role.

关 键 词：胃肿瘤 脱噬作用 丙谷胺 受体 缩胆囊素 

分 类 号：R735.2[医药卫生—肿瘤]