南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2015年
6期
883-887
,共5页
乳腺癌细胞%氯尼达明%三磷酸腺苷%葡萄糖调节蛋白78%凋亡抑制蛋白家族
乳腺癌細胞%氯尼達明%三燐痠腺苷%葡萄糖調節蛋白78%凋亡抑製蛋白傢族
유선암세포%록니체명%삼린산선감%포도당조절단백78%조망억제단백가족
breast cancer cells%lonidamine%ATP%GRP78%inhibitor of apoptosis protein
目的：探讨氯尼达明诱导人乳腺癌MCF-7细胞凋亡的作用及其可能机制。方法 MTT法及集落形成实验检测氯尼达明对乳腺癌细胞MCF-7增殖的抑制作用；PI/Annexin-V双染检测细胞凋亡；ATP检测试剂盒检测细胞内ATP水平；Western blot检测葡萄糖调节蛋白78（GRP78）、凋亡抑制蛋白家族cIAP1及caspase-8蛋白的表达。结果 MTT结果显示，50~250 mmol · L-1氯尼达明可抑制MCF-7细胞的增殖活性，且呈浓度和时间依赖性。集落形成实验同样证明了上述结果。PI/Annexin-V双染结果表明，随着氯尼达明浓度的增加，MCF-7细胞的凋亡率也随之增加。50、150和250 mmol · L-1氯尼达明作用于MCF-7细胞5 h后，细胞内ATP水平与对照组相比分别为80.67%、62.78%和30.73%。Western blot检测发现，随着氯尼达明作用时间的延长， GRP78蛋白表达上调，cIAP1蛋白表达下调，同时增强了caspase-8的活性。结论氯尼达明可以抑制乳腺癌细胞MCF-7的增殖活性，诱导其产生凋亡，其机制可能与减少细胞内ATP的产生诱导内质网应激反应，并下调cIAP表达及增强caspase-8的活性有关。
目的：探討氯尼達明誘導人乳腺癌MCF-7細胞凋亡的作用及其可能機製。方法 MTT法及集落形成實驗檢測氯尼達明對乳腺癌細胞MCF-7增殖的抑製作用；PI/Annexin-V雙染檢測細胞凋亡；ATP檢測試劑盒檢測細胞內ATP水平；Western blot檢測葡萄糖調節蛋白78（GRP78）、凋亡抑製蛋白傢族cIAP1及caspase-8蛋白的錶達。結果 MTT結果顯示，50~250 mmol · L-1氯尼達明可抑製MCF-7細胞的增殖活性，且呈濃度和時間依賴性。集落形成實驗同樣證明瞭上述結果。PI/Annexin-V雙染結果錶明，隨著氯尼達明濃度的增加，MCF-7細胞的凋亡率也隨之增加。50、150和250 mmol · L-1氯尼達明作用于MCF-7細胞5 h後，細胞內ATP水平與對照組相比分彆為80.67%、62.78%和30.73%。Western blot檢測髮現，隨著氯尼達明作用時間的延長， GRP78蛋白錶達上調，cIAP1蛋白錶達下調，同時增彊瞭caspase-8的活性。結論氯尼達明可以抑製乳腺癌細胞MCF-7的增殖活性，誘導其產生凋亡，其機製可能與減少細胞內ATP的產生誘導內質網應激反應，併下調cIAP錶達及增彊caspase-8的活性有關。
목적：탐토록니체명유도인유선암MCF-7세포조망적작용급기가능궤제。방법 MTT법급집락형성실험검측록니체명대유선암세포MCF-7증식적억제작용；PI/Annexin-V쌍염검측세포조망；ATP검측시제합검측세포내ATP수평；Western blot검측포도당조절단백78（GRP78）、조망억제단백가족cIAP1급caspase-8단백적표체。결과 MTT결과현시，50~250 mmol · L-1록니체명가억제MCF-7세포적증식활성，차정농도화시간의뢰성。집락형성실험동양증명료상술결과。PI/Annexin-V쌍염결과표명，수착록니체명농도적증가，MCF-7세포적조망솔야수지증가。50、150화250 mmol · L-1록니체명작용우MCF-7세포5 h후，세포내ATP수평여대조조상비분별위80.67%、62.78%화30.73%。Western blot검측발현，수착록니체명작용시간적연장， GRP78단백표체상조，cIAP1단백표체하조，동시증강료caspase-8적활성。결론록니체명가이억제유선암세포MCF-7적증식활성，유도기산생조망，기궤제가능여감소세포내ATP적산생유도내질망응격반응，병하조cIAP표체급증강caspase-8적활성유관。
Objective To investigate the effect of lonidamine on apoptosis of human breast carcinoma cells MCF-7 and the possible mechanisms. Methods MTT assay and colony-forming assay were used to evaluate the growth inhibition induced by lonidamine in breast cancer MCF-7 cells. PI/Annexin-V staining was used to detect the apoptotic cells. The ATP levels in the cells were detected using an ATP assay kit. The expression of glucose regulated protein 78 (GRP78), inhibitor of apoptosis protein (cIAP1) and caspase-8 were analyzed with Western blotting. Results MTT assay and colony-forming assay showed that 50-250 mmol/L lonidamine caused a time- and concentration-dependent inhibition of MCF-7 cell proliferation. Exposure to increased concentrations of lonidamine resulted in significantly increased apoptosis rate in MCF-7 cells. In MCF-7 cells treated with 50, 150 and 250 mmol/L lonidamine for 5 h, the intracellular ATP levels were lowered to 80.67%, 62.78%and 30.73%of the control level, respectively. Western blotting showed that lonidamine up-regulated the expression of GRP78, down-regulated the expression of cIAP1 and promoted caspase-8 activation as the treatment time extended. Conclusion Lonidamine can inhibit the proliferation and induce apoptosis in MCF-7 cells, and these effects are probably mediated by reducing ATP level, inducing endoplasmic reticulum stress response, down-regulating cIAP1, and promoting caspase-8 activation in the cells.
