摘要
目的:研究雷丸蛋白对人胃癌细胞MC-4凋亡的诱导作用。方法:以聚乙烯吡咯烷酮(PVP)为溶剂提取雷丸饮片蛋白质,BCA法测定蛋白浓度后以DMEM培养液稀释到180mg.mL-1,120mg.mL-1,90mg.mL-1,60mg.mL-1和45 mg.mL-1,体外作用于胃癌细胞MC-4,37℃、5%CO2条件下培养18h后,光镜检查作用结果。以Hoechst 33258染色后于荧光显微镜下观察细胞凋亡情况,以Annexin V-PI双染色后以流式细胞仪检查细胞的死亡和凋亡情况。结果:随着蛋白质用量的增加,MC-4细胞死亡率与剂量间呈现量效关系,180mg.mL-1时达最高值29.6%。凋亡率和剂量间呈现较好的剂量-效应曲线,120mg.mL-1时凋亡率最高,可达29.4%,聚乙烯吡咯烷酮无作用。结论:PVP荷载雷丸蛋白对人胃癌细胞MC-4有很好的杀伤作用和诱导凋亡作用,杀伤机理和凋亡机理有待深入研究。
Objective：To observe the apoptosis of human gastric epithelial MC-4 cell induced by proteins of Omphalia lapideacens conjugation to polyvinylpyrrolidone（PVP）.Methods：Proteins extracted from Omphalia lapideacens in PVP solution were diluted with DMEM medium after be determined concentration by BCA method.The MC-4 cell were exposed to 180mg·mL-1,120 mg·mL-1,90mg·mL-1,60mg·mL-1 and 45 mg·mL-1 respectively for 18 hr in a humidified atmosphere of 95% air plus 5%CO2 at 37℃.The morphological changes were observed under microscope at 200×magnification.Hoechst 33258 staining was performed to evaluate chromosome condensation.The apoptotic cells were determined by Annexin V-PI double staining followed by flow cytometric analysis.Results： There was a significant correlation between the protein concentration and the percentage of dead cells and apoptosis cells.The peak value of mortality rate and apoptosis rate was 29.6% and 29.4% respectively in 180mg·mL-1 and 120mg·mL-1 concentration at 18 hr.However,PVP had no effect on the MC-4 cell.Conclusion：The proteins of Omphalia lapideacens in conjugation with PVP can elicit apoptosis and damage the viability of MC-4 cell.But the mechanism was needed further study to demonstrate.
出处
《中华中医药学刊》
CAS
2011年第6期1296-1298,共3页
Chinese Archives of Traditional Chinese Medicine
基金
浙江省自然科学基金资助项目(Y207765)
浙江省教育厅科研项目(20050817)
关键词
雷丸
蛋白
聚乙烯吡咯烷酮
细胞凋亡
Omphalia lapideacens
protein
polyvinylpyrrolidone
apoptosis
