中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2014年
21期
64-67
,共4页
肖芬%戈伟%郑永法%吴方红
肖芬%戈偉%鄭永法%吳方紅
초분%과위%정영법%오방홍
吗啡%HGC27细胞%Caspase-3%p16%bcl-2%bax
嗎啡%HGC27細胞%Caspase-3%p16%bcl-2%bax
마배%HGC27세포%Caspase-3%p16%bcl-2%bax
Morphine%HGC27 cell%Caspase-3%p16%bcl-2%bax
目的：探讨吗啡对人胃癌细胞系HGC27细胞增殖的影响及其机制。方法体外培养的HGC27细胞，用不同浓度吗啡处理24、48、72 h，四甲基偶氮唑蓝(MTT)法测定其对HGC27细胞增殖的影响；倒置显微镜下观察细胞的形态学变化；实验组用吗啡(浓度分别为0.05、0.1、0.2μmol/L)处理HGC27细胞48 h，对照组加入不含药物的培养基，分别采用流式细胞仪检测细胞凋亡、分光光度计检测细胞凋亡蛋白酶-3(Casepase-3)相对活性、west-ern blot法检p16、bcl-2、bax基因表达情况。结果吗啡(浓度0.025～0.4μmol/L)能抑制HGC27细胞的增殖，呈剂量和时间依赖性；倒置显微镜下可观察到典型的细胞凋亡形态；吗啡(浓度分别为0.05、0.1、0.2μmol/L)作用HGC27细胞48 h后，细胞凋亡率分别为9.4%、11.5%、21.4%，而对照组凋亡率仅为2.2%；Casepase-3相对活性显著增加，处理后Casepase-3相对活性分别为(1.32依0.08)、(1.85依0.06)和(2.45依0.07)，对照组为(0.92依0.07)；p16和bax蛋白表达量升高，而bcl-2蛋白表达量降低，呈剂量依赖性。结论吗啡能抑制HGC27细胞的增殖并诱导其凋亡，其作用机制可能与上调p16表达，进而引起bax蛋白表达量升高，而bcl-2蛋白表达量降低，并增加细胞Casepase-3活性有关。
目的：探討嗎啡對人胃癌細胞繫HGC27細胞增殖的影響及其機製。方法體外培養的HGC27細胞，用不同濃度嗎啡處理24、48、72 h，四甲基偶氮唑藍(MTT)法測定其對HGC27細胞增殖的影響；倒置顯微鏡下觀察細胞的形態學變化；實驗組用嗎啡(濃度分彆為0.05、0.1、0.2μmol/L)處理HGC27細胞48 h，對照組加入不含藥物的培養基，分彆採用流式細胞儀檢測細胞凋亡、分光光度計檢測細胞凋亡蛋白酶-3(Casepase-3)相對活性、west-ern blot法檢p16、bcl-2、bax基因錶達情況。結果嗎啡(濃度0.025～0.4μmol/L)能抑製HGC27細胞的增殖，呈劑量和時間依賴性；倒置顯微鏡下可觀察到典型的細胞凋亡形態；嗎啡(濃度分彆為0.05、0.1、0.2μmol/L)作用HGC27細胞48 h後，細胞凋亡率分彆為9.4%、11.5%、21.4%，而對照組凋亡率僅為2.2%；Casepase-3相對活性顯著增加，處理後Casepase-3相對活性分彆為(1.32依0.08)、(1.85依0.06)和(2.45依0.07)，對照組為(0.92依0.07)；p16和bax蛋白錶達量升高，而bcl-2蛋白錶達量降低，呈劑量依賴性。結論嗎啡能抑製HGC27細胞的增殖併誘導其凋亡，其作用機製可能與上調p16錶達，進而引起bax蛋白錶達量升高，而bcl-2蛋白錶達量降低，併增加細胞Casepase-3活性有關。
목적：탐토마배대인위암세포계HGC27세포증식적영향급기궤제。방법체외배양적HGC27세포，용불동농도마배처리24、48、72 h，사갑기우담서람(MTT)법측정기대HGC27세포증식적영향；도치현미경하관찰세포적형태학변화；실험조용마배(농도분별위0.05、0.1、0.2μmol/L)처리HGC27세포48 h，대조조가입불함약물적배양기，분별채용류식세포의검측세포조망、분광광도계검측세포조망단백매-3(Casepase-3)상대활성、west-ern blot법검p16、bcl-2、bax기인표체정황。결과마배(농도0.025～0.4μmol/L)능억제HGC27세포적증식，정제량화시간의뢰성；도치현미경하가관찰도전형적세포조망형태；마배(농도분별위0.05、0.1、0.2μmol/L)작용HGC27세포48 h후，세포조망솔분별위9.4%、11.5%、21.4%，이대조조조망솔부위2.2%；Casepase-3상대활성현저증가，처리후Casepase-3상대활성분별위(1.32의0.08)、(1.85의0.06)화(2.45의0.07)，대조조위(0.92의0.07)；p16화bax단백표체량승고，이bcl-2단백표체량강저，정제량의뢰성。결론마배능억제HGC27세포적증식병유도기조망，기작용궤제가능여상조p16표체，진이인기bax단백표체량승고，이bcl-2단백표체량강저，병증가세포Casepase-3활성유관。
Objective To investigate the effect of morphine on cell proliferation in gastric cancer line HGC27 cells and discuss its possible mechanism. Methods HGC27 cells were cultured in vitro. After treatment by morphine at different concentrations respectively at different time, the cell survival was determined by the MTT method. The changes of cell morphology were observed by inverted microscope. Apoptosis was detected by flow cytometry. The relative activity of caspase-3 was monitored by spectrophotometer. The changes of protein expression of p16, bcl-2 and bax were detected by western blot. Results From the data of MTT, the cell proliferation of human gastricl cancer HGC27 cells was inhib-ited by morphine in a dose-dependent and time-dependent manner. Typical apoptosis morphology of HGC27 cells was observed by inverted microscope. Flow cytometry assays showed that morphine significantly induced apoptosis in HGC27 cells. After treated with morphine, the apoptosis rate of HGC27 cells was 9.4%, 11.5% and 21.4%respectively, which showed an obvious concentration-effect relationship. The relative activity of caspase-3 of morphine group was (1.32±0.08), (1.85±0.06) and (2.45±0.07) respectively, which of control group was (0.92±0.07). The data of western blot showed that morphine up-regulated p16 and bax and down- regulated bcl-2 expression in a dose-dependent manner. Conclusion Morphine can inhibit the proliferation of HGC27 cells and induce apoptosis, and the mechanism of mor-phine on apoptosis may be related to the up-regulation of p16 expression, thus results up-regulation of bax and down-regulation of bcl-2, as well as the increase of relative activity of caspase-3.
