CAJ | 학술논문

目的观察大蒜素主要成分二烯丙基硫(DAS)诱导人成骨肉瘤细胞株MG63的增殖抑制作用及其对血管内皮生长因子(VEGF)表达的影响,并探讨其机制.方法以0、5、10、20、40、80g/L的DAS作用于MG63细胞24、48、72 h,噻唑蓝(MTT)比色法检测细胞增殖活性,利用倒置相差显微镜及Hoechst33258荧光染色法观察细胞形态学改变;流式细胞术检测0、10、20、40 g/L的DAS作用于MG63 48 h后细胞周期分布及凋亡率;半定量逆转录-聚合酶链反应(RT-PCR)和Western blot分别检测VEGF-mRNA和VEGF的表达水平.结果大蒜素能明显抑制MG63细胞增殖,并呈时间-效应和剂量-效应变化.当大蒜素浓度＞10 g/L时,药物对MG63细胞增殖的抑制作用明显强于空白对照组(P＜0.05).光镜下可见肿瘤细胞皱缩变圆,部分细胞脱壁悬浮.10、20、40g/L作用于MG63细胞48 h后细胞凋亡率分别为(4.7±1.4)%、(18.6±0.7)%、(24.9±1.1)%,均明显高于对照组48 h自然凋亡率(1.70±0.36)%(P＜0.05).DAS可使MG63细胞中VEGF-mRNA及蛋白含量明显减少,并呈浓度依赖性.结论 DAS能通过抑制人成骨肉瘤细胞株MG63血管生成诱导肿瘤细胞的凋亡.
목적 관찰대산소주요성분이희병기류(DAS)유도인성골육류세포주MG63적증식억제작용급기대혈관내피생장인자(VEGF)표체적영향,병탐토기궤제.방법 이0、5、10、20、40、80g/L적DAS작용우MG63세포24、48、72 h,새서람(MTT)비색법검측세포증식활성,이용도치상차현미경급Hoechst33258형광염색법관찰세포형태학개변;류식세포술검측0、10、20、40 g/L적DAS작용우MG63 48 h후세포주기분포급조망솔;반정량역전록-취합매련반응(RT-PCR)화Western blot분별검측VEGF-mRNA화VEGF적표체수평.결과 대산소능명현억제MG63세포증식,병정시간-효응화제량-효응변화.당대산소농도＞10 g/L시,약물대MG63세포증식적억제작용명현강우공백대조조(P＜0.05).광경하가견종류세포추축변원,부분세포탈벽현부.10、20、40g/L작용우MG63세포48 h후세포조망솔분별위(4.7±1.4)%、(18.6±0.7)%、(24.9±1.1)%,균명현고우대조조48 h자연조망솔(1.70±0.36)%(P＜0.05).DAS가사MG63세포중VEGF-mRNA급단백함량명현감소,병정농도의뢰성.결론 DAS능통과억제인성골육류세포주MG63혈관생성유도종류세포적조망.
Objective To investigate the effect of diallyl sulfide (DAS) on the growth and vascular endothelial growth factor (VEGF) expression of human MG63 cells in vitro, and to explore the curative possibility of osteosarcoma with DAS. Methods The effect of DAS at the concentrations of 0, 10, 20, 40 g/L on growth and proliferation was assessed by methylthiazol tetrazolium (MTT) assay. The morphology of the cells was examined under the inverted phase contrast microscopy and Hoechst 33258 fluorescent staining. The apoptosis was detected by annexin-V/PI double-labeled cytometry after treated with DAS at different concentrations of 0, 10, 20, 40 g/L for 48 h. The effect of DAS on the cell cycle of MG63 cells was studied by flow cytometry. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to analyze the changes of VEGF mRNA and protein levels. Results Proliferation of MG63cells was significantly inhibited by DAS in a time- and dose-dependent fashion. As compared with the control group, the DAS over the concentration of 10 g/L significantly inhibited the proliferation of MG63 cells (P＜0.05). MG63 cells grew round and small obviously after being treated with DAS under the inverted microscopy. The apoptotic rate was(4.7±1.4)%, (18.6±0.7)%, (24.9±1.1)% after the cells were treated with DAS at the concentrations of 10, 20, 40 g/L for 48 h, respectively, which was significantly different from that in the control group (1.70±0.36)%, P＜0.05. Allicin reduced the secretion of VEGF and inhibited the level of VEGF mRNA expression in a dose-dependent manner. Conclusion DAS can inhibit the proliferation of MG63 cells in vitro. The antitumor effects of allicin may be related to down-regulation of the expression of VEGF mRNA.