CAJ | 학술논문

目的：探讨毒毛旋花子阿洛糖苷诱导胃腺癌细胞 SGC -7901凋亡作用及其机制。方法将人胃腺癌细胞 SGC -7901分为对照组、0.125μg/ ml 组、0.250μg/ ml 组、0.500μg/ ml 组、1.000μg/ ml 组，四甲基偶氮唑盐微量酶反应比色法（MTT）检测毒毛旋花子阿洛糖苷对胃腺癌细胞 SGC -7901的生长抑制作用，Hoechst 染色和 Annexin V - FITC/碘化丙啶（PI）流式细胞术检测细胞凋亡情况，JC -1法检测线粒体膜电位变化，Western blotting 法检测线粒体途径相关蛋白细胞色素 C、Caspase -3和 Caspase -9的表达。结果 MTT 结果显示，24 h 和48 h 时不同组胃腺癌细胞 SGC -7901吸光度（A）、生长抑制率比较，差异均有统计学意义（P <0.05）；组间两两比较胃腺癌细胞 SGC -7901 A 值依次降低、生长抑制率依次升高（P <0.05）。荧光显微镜下观察发现，对照组细胞核显示为蓝荧光且较均匀，0.500μg/ ml组处理12 h 后细胞核呈紧密较亮荧光体，颜色较白。不同组胃腺癌细胞 SGC -7901总相对死亡率比较，差异有统计学意义（P <0.05）；组间两两比较胃腺癌细胞 SGC -7901总相对死亡率依次升高（P <0.05）。JC -1染色结果显示，毒毛旋花子阿洛糖苷处理胃腺癌细胞 SGC -790112 h 后，0.125μg/ ml 组、0.250μg/ ml 组、0.500μg/ml 组显示绿色荧光，随毒毛旋花子阿洛糖苷浓度的增加绿色荧光增强。Western blotting 法显示，随着细胞凋亡的发生，细胞色素 C 从线粒体中释放到细胞质中，同时，Caspase -3和 Caspase -9也被激活并发生剪切活化，活化片段表达增加。结论毒毛旋花子阿洛糖苷通过线粒体凋亡途径诱导体外培养的人胃腺癌细胞 SGC -7901凋亡。
목적：탐토독모선화자아락당감유도위선암세포 SGC -7901조망작용급기궤제。방법장인위선암세포 SGC -7901분위대조조、0.125μg/ ml 조、0.250μg/ ml 조、0.500μg/ ml 조、1.000μg/ ml 조，사갑기우담서염미량매반응비색법（MTT）검측독모선화자아락당감대위선암세포 SGC -7901적생장억제작용，Hoechst 염색화 Annexin V - FITC/전화병정（PI）류식세포술검측세포조망정황，JC -1법검측선립체막전위변화，Western blotting 법검측선립체도경상관단백세포색소 C、Caspase -3화 Caspase -9적표체。결과 MTT 결과현시，24 h 화48 h 시불동조위선암세포 SGC -7901흡광도（A）、생장억제솔비교，차이균유통계학의의（P <0.05）；조간량량비교위선암세포 SGC -7901 A 치의차강저、생장억제솔의차승고（P <0.05）。형광현미경하관찰발현，대조조세포핵현시위람형광차교균균，0.500μg/ ml조처리12 h 후세포핵정긴밀교량형광체，안색교백。불동조위선암세포 SGC -7901총상대사망솔비교，차이유통계학의의（P <0.05）；조간량량비교위선암세포 SGC -7901총상대사망솔의차승고（P <0.05）。JC -1염색결과현시，독모선화자아락당감처리위선암세포 SGC -790112 h 후，0.125μg/ ml 조、0.250μg/ ml 조、0.500μg/ml 조현시록색형광，수독모선화자아락당감농도적증가록색형광증강。Western blotting 법현시，수착세포조망적발생，세포색소 C 종선립체중석방도세포질중，동시，Caspase -3화 Caspase -9야피격활병발생전절활화，활화편단표체증가。결론독모선화자아락당감통과선립체조망도경유도체외배양적인위선암세포 SGC -7901조망。
Objectjve To explore the apoptosis - inducing effect of Strophalloside on gastric adenocarcinoma cells type SGC - 7901 and its mechanism. Methods The human gastric adenocarcinoma cells type SGC - 7901 were divided into control group,0. 125 μg/ ml group,0. 250 μg/ ml group,0. 500 μg/ ml group,and 1. 000 μg/ ml group. The growth inhibition effect of Strophalloside on gastric adenocarcinoma cells type SGC - 7901 was detected by MTT assay,and the apoptosis was measured by Annexin V - FITC/ PI flow cytometry and Hoechst staining. The changes of mitochondrial membrane potential were examined by JC - 1 kit. The expressions of mitochondrial pathway - related protein cytochrome C,Caspase - 3 and Caspase - 9 were detected by Western blotting analysis. Results According to MTT assay results,there were significant differences in value A and rate of growth inhibition of gastric adenocarcinoma cells type SGC - 7901 among different groups at 24 h and 48 h after treatment, respectively(P < 0. 05);comparison of the above two indicators between groups showed that value A of gastric adenocarcinoma cells type SGC - 7901 decreased sequentially,and rate of growth inhibition increased sequentially(P < 0. 05). By fluorescence microscope,the cell nucleus of control group showed homogeneous blue fluorescence,the cell nucleus of 0. 500 μg/ ml group showed compact bright fluorescence with white color. There was significant difference in total relative mortality of gastric adenocarcinoma cells type SGC - 7901 among different groups( P < 0. 05);comparison of the above indicator between groups showed that total relative mortality of gastric adenocarcinoma cells type SGC - 7901 increased sequentially( P < 0. 05 ). According to results of JC - 1 staining,gastric adenocarcinoma cells type SGC - 7901 showed green fluorescence 12 h after Strophalloside treatment,and fluorescence intensity increased as concentration of Strophalloside rised. According to Western blotting results,cytochrome C was released from mitochondria into cytoplasm C,and Caspase - 3 and Caspase - 9 were activated,shear and activation occurred,the expression level of activated fragments increased. Conclusjon Strophalloside can induce apoptosis of gastric adenocarcinoma cells type SGC - 7901 through the mitochondria - mediated pathway.