CAJ | 학술논문

目的：探讨胃泌素对人大肠癌细胞株SW480增殖、凋亡的影响，以及P38-丝裂素活化蛋白激酶（ MAPK）信号转导通路的关系。方法：体外培养大肠癌SW480细胞株，将细胞分为对照组、胃泌素组、丙谷胺组和胃泌素＋丙谷胺组，对照组不加药，其余组加不同浓度的药物处理。运用MTT法观察细胞凋亡情况，确立5-肽胃泌素和丙谷胺处理SW480细胞的最佳浓度；流式细胞术检测各组细胞增殖指数的变化；qRT-PCR检测大肠癌SW480细胞株胃泌素受体／胆囊收缩素-B受体（ CCK-2R） mRNA表达情况；Western blot 检测P38蛋白表达及其磷酸化水平。采用方差分析和SNK-q检验进行统计学分析。结果：SW480细胞中存在CCK-2R mRNA表达。胃泌素在6.25～200.00 mg／L范围内能抑制大肠癌SW480细胞的凋亡，且当浓度达12.50 mg／L时为最佳浓度（ P＜0.05）；单独丙谷胺对大肠癌SW480细胞凋亡无明显影响（ P＞0.05），但丙谷胺在8.00～256.00 mg／L范围内能明显拮抗胃泌素抑制SW480细胞的凋亡作用，其最佳浓度为16.00 mg／L （ P＜0.05）。胃泌素（12.50 mg／L）组细胞增殖指数为（34.56±1.41）％，显著高于对照组的（28.74±1.61）％和胃泌素（12.50 mg／L）＋丙谷胺（16.00 mg／L）组的（28.72±1.78）％（P＜0.05），而丙谷胺（16.00 mg／L）组细胞增殖指数为（27.75±2.29）％，与对照组比较差异无统计学意义（P＞0.05）；P38蛋白及其磷酸化在胃泌素组（12.50 mg／L）中表达水平低于对照组（P＜0.01），也低于胃泌素（12.50 mg／L）＋丙谷胺（16.00 mg／L）组（ P＜0.01）。结论：胃泌素可以抑制大肠癌细胞株SW480的凋亡、促进增殖，其作用可能是通过抑制P38及其磷酸化水平来实现的，但能够被其受体拮抗剂丙谷胺抑制。
목적：탐토위비소대인대장암세포주SW480증식、조망적영향，이급P38-사렬소활화단백격매（ MAPK）신호전도통로적관계。방법：체외배양대장암SW480세포주，장세포분위대조조、위비소조、병곡알조화위비소＋병곡알조，대조조불가약，기여조가불동농도적약물처리。운용MTT법관찰세포조망정황，학립5-태위비소화병곡알처리SW480세포적최가농도；류식세포술검측각조세포증식지수적변화；qRT-PCR검측대장암SW480세포주위비소수체／담낭수축소-B수체（ CCK-2R） mRNA표체정황；Western blot 검측P38단백표체급기린산화수평。채용방차분석화SNK-q검험진행통계학분석。결과：SW480세포중존재CCK-2R mRNA표체。위비소재6.25～200.00 mg／L범위내능억제대장암SW480세포적조망，차당농도체12.50 mg／L시위최가농도（ P＜0.05）；단독병곡알대대장암SW480세포조망무명현영향（ P＞0.05），단병곡알재8.00～256.00 mg／L범위내능명현길항위비소억제SW480세포적조망작용，기최가농도위16.00 mg／L （ P＜0.05）。위비소（12.50 mg／L）조세포증식지수위（34.56±1.41）％，현저고우대조조적（28.74±1.61）％화위비소（12.50 mg／L）＋병곡알（16.00 mg／L）조적（28.72±1.78）％（P＜0.05），이병곡알（16.00 mg／L）조세포증식지수위（27.75±2.29）％，여대조조비교차이무통계학의의（P＞0.05）；P38단백급기린산화재위비소조（12.50 mg／L）중표체수평저우대조조（P＜0.01），야저우위비소（12.50 mg／L）＋병곡알（16.00 mg／L）조（ P＜0.01）。결론：위비소가이억제대장암세포주SW480적조망、촉진증식，기작용가능시통과억제P38급기린산화수평래실현적，단능구피기수체길항제병곡알억제。
Objective:To investigate the effects of gastrin on the proliferation and apoptosis of human large intestinal carcinoma cell line SW 480 and the relationship of signal transduction pathway in p38 mitogen-activated protein kinases(P38-MAPK).Methods:SW 480 cell line of human large intestinal cancer was in vitro incubated and divided into groups of control ,gastrin use,proglumide intervention and gastrin +proglumide.The control group were free of medication,and the remaining groups were treated with corresponding drugs in different dosage .MTT reduction assay was performed to detect the cell ap-optosis status and determine the optimal dosage of pentagstrin and proglumide affecting SW480 cell line proliferation.Flow cytometry was used to estimate the proliferation index in each group,and qRT-PCR was performed to detect the expression of gastrin receptor (CCK-2R) mRNA in SW480 cells.Western blot was performed to examine the expression of P38 protein and its phosphorylation levels.All results were analyzed with χ2 test and SNK-q test.Results:qRT-PCR detection showed that CCK-2R mRNA was expressed in the SW480 cell line.Pentagastrin inhibited SW480 cell apoptosis in a dose ranging from 6.25 to 200 mg/L,and the optimal dosage was 12.50 mg/L(P<0.05).Proglumide was capable of inhibiting the cell proliferation in dosage of 8.00 -256.00 mg/L,and the optimal effect was in dose of 16.00 mg/L(P<0.05).The proliferation index in pentagastrin group was (34.56 ±1.41)%,which was significantly higher than that of control group(28.74 ±1.61) % and combined treatment group(28.72 ±1.78)%(P<0.05),and the index differ-ence was not significant between proglumide group and the controls(P>0.05).P38 protein expression and its phosphorylation level were lower in penta-gastrin group as compared with the controls and combination treatment group(P<0.01).Conclusion:Gastrin can inhibit the apoptosis of human large in-testinal cancer cell line SW480 and improve the cell proliferation.This possible mechanism may be associated with suppressed p38 protein and its phospho-rylation level due to inhibition of proglumide,a gastrin receptor antagonist.