文档介绍：半夏蛋白抗肿瘤活性组分的提取分离
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作者:付芸,黄必胜,李娟,田代志,陈科力
【摘要】目的分离半夏蛋白的抗肿瘤活性组分,研究该半夏蛋白组分的抗肿瘤作用。方法用凝胶柱分离纯化半夏粗总蛋白,采用四氮唑蓝比色法(MTT法)观察半夏蛋白对肝癌Bel-7402细胞增殖的抑制,应用流式细胞术(FCM)检测其对细胞凋亡的影响。结果半夏粗总蛋白中30%硫酸铵沉淀部分、60%硫酸铵沉淀部分对肿瘤抑制的量效关系不明显,30% mol/ mol/L NaCl的洗脱峰呈现较好的量效关系,且对Bel-7402细胞的抑制作用与空白对照组有显著性差异。30%硫酸铵沉淀蛋白有促进Bel-7402细胞凋亡的作用。结论半夏蛋白中30%硫酸铵沉淀部分对Bel-7402细胞生长具有明显抑制作用及促进Bel-7402细胞凋亡的作用。
【关键词】半夏;Bel-7402细胞;抗肿瘤蛋白组分;抗肿瘤活性测定
Abstract:Objective To isolate the anti-tumor protein groups from Pinellia ternata rhizome and investigate the anti-tumor activity of these protein groups on Bel-7402 cell. Methods The total raw protein was isolated with sepharose column chromatography. Methyl-thiazolyl-tetrazolinm (MTT) was used to analyze the effect of Pinellia ternata Protein on inhibiting growth of tumor cells. Apoptosis was detected by flow cytometry (FCM). Results The 30% and 60% (NH4)2SO4 deposition part of total proteins from Pinellia ternata rhizome have no certain relationship between quantity and inhibitory action, but protein peak 3 eluted with mol/L or mol/L NaCl from 30% (NH4)2SO4 deposition part showed a effect of concentration depending. FCM analysis showed that the protein of 30% (NH4)2SO4 deposition part could induce apoptosis. Conclusion The 30% (NH4)2SO4 deposition part of total proteins from Pinellia ternata rhizome could significan