丙谷胺和NS-398协同诱导胃癌细胞凋亡的研究

Proglumide and NS-398 synergistically induces apoptosis in gastric cancer cell

目的:探讨胃泌素受体拮抗剂丙谷胺和特异性环氧化酶(COX)-2抑制剂NS-398对人胃腺癌细胞株MKN-45细胞凋亡的调控作用及其机制。方法:MKN-45细胞常规培养于含10%胎牛血清的RPMI1640培养液中,待细胞长至亚单层后加丙谷胺(5mmol/L)和(或)NS-398(0.01mmol/L),连续培养48h。采用流式细胞仪检测细胞凋亡率,RT-PCR法测定凋亡抑制基因bcl-2mRNA表达。结果:丙谷胺组、NS-398组和联合用药组的细胞凋亡率分别为(24.72±3.19)%、(26.69±3.35)%和(36.11±4.57)%,显著高于对照组(1.57±0.55)%(P<0.01),联合用药组的细胞凋亡率明显高于丙谷胺或NS-398单一用药组[(36.11±4.57)%vs(24.72±3.19)%和(26.69±3.35)%,P<0.05]。丙谷胺和NS-398显著下调bcl-2mRNA在MKN-45细胞的表达(P<0.05)。结论:丙谷胺、NS-398通过下调胃癌细胞bcl-2基因表达而诱导细胞凋亡,两者联合使用具有协同作用。

Objective ： To investigate the regulative effects and their mechanism of a gastrin receptor antagonist proglumide and a specific COX-2 inhibitor NS-398 on the apoptosis in a gastric cancer cell line, MKN-45. Methods： MKN-45 cells were routinely cultured in RPMI1640 medium supplemented with 10% heat-inactivated fetal calf serum. Subconfluent cell cultures were treated with either proglumide at a final concentration of 5 mmol/L, or NS-398 at a final concentration of 0.01 mmol/L, or proglumide in combination with NS-398 for 48 h. Flow cytometric analysis was used to detect the apoptosis status of MKN-45 cells. The expression of apoptosis-inhibited gene bcl-2 was assessed by RT-PCR. Results： Apoptosis rate was （24.72 ± 3.19）%, （26.69 ± 3.35）% and （36.11 ± 4.57）% in the MKN-45 cells treated with proglumid, NS-398, and combination of the two agents, respectively, which were significantly higher than that in the control group [ （ 1.57 ± 0.55 ） %, P 〈 0.01 ]. The apoptosis rates in the cells treated with proglumid and NS-398 were much greater than that treated with proglumid or NS-398 applied singly （P 〈 0.05 ）. Proglumide and NS-398 remarkably reduced the expression of bcl-2 mRNA （P 〈 0.05）. Conclusion： Both proglumide and NS-398 are able to induce the apoptosis in MKN-45 cells. This apoptosis may be mediated by down-expression of apoptosis-inhibited gene bcl-2. Costimulation with proglumide and NS-398 synergistically induces apoptosis in gastric cancer cells.