摘要
目的研究微量元素锂（ＬｉＣｌ）在体外对肿瘤细胞系凋亡的诱导作用。方法在细胞培养体系中加入Ｌｉｃｌ进行诱导，以流式细胞分析、ＤＮＡ电泳分析及细胞形态学改变为指标检测凋亡，并用ｍＲＮＡ狭缝杂交和Ｗｅｓｔｅｒｎｂｌｏｔ测定凋亡相关基因的改变。加入大分子合成抑制剂或第二信使相关药物观察对锂诱导凋亡的影响。结果ＬｉＣｌ可以诱导人髓性细胞白血病细胞系ＨＬ－６０、人单核细胞白血病细胞系Ｕ９３７和人卵巢癌细胞系Ａ２７８０产生凋亡。蛋白合成抑制剂放线菌酮（Ｃｙｃｌｏｈｅｘｉｍｉｄｅ，ＣＨＸ）或ＲＮＡ合成抑制剂放线菌素Ｄ（ＡｃｔｉｎｏｍｙｃｉｎＤ，ＡＣＤ）均不能抑制ＬｉＣｌ诱导ＨＬ－６０细胞凋亡，提示该过程中不需要蛋白重新合成。此外，升高胞内ｃＡＭＰ水平的药物霍乱毒素（ｃｈｏｌｅｒａｔｏｘｉｎ，ＣＴ）抑制ＬｉＣｌ诱导ＨＬ－６０细胞凋亡；ＰＫＣ的抑制剂Ｄ－ｓｐｈｉｇｏｓｉｎｅ（ＤＳ）能诱导ＨＬ－６０细胞产生一定水平的凋亡，与锂共同作用未测得加合效应；胞内钙离子螯合剂Ｎｉｃａｒｄｉｐｉｎｅ或胞外钙离子螯合剂ＥＧＴＡ对ＬｉＣｌ诱导ＨＬ－６０凋亡均无影响。锂处理ＨＬ－６０细胞引起ｂｃｌ－２、ｃ－ｍｙｃ表达水平的降低；处理Ｕ９３７细胞引?
The induction of apoptosis by trace element lithium in three human tumor cell lines was studied.Lithium(LiCl) was found to be able to induce apoptosis in HL60,U937 and A2780 in vitro. When HL60 cells were pretreated with actinomycin D(ACD) or cycloheximide (CHX) then incubated with LiCl,neither ACD nor CHX could inhibit apoptosis induced by LiCl,indicating that protein synthesis is not required for these cells to undergo apoptosis.Cholera toxin(CT) which can enhance intracellular cAMP level resulted in inhibition of apoptosis induced by LiCl.Dsphigosine(DS),a protein kinase C inhibitor,alone could cause apoptosis in HL60.However,as DS in combination with lithium the additive effect could not be obtained.Our data also showed that neither nicardipine nor EGTA had any influence on apoptotic induction by LiCl.These results reveal that the induction of apoptosis of HL60 by LiCl is associat with cAMP and PKC,but is independent on Ca2+.Furthermore,the effects of lithium on the expression of apoptosis associated genes bcl2,cmyc and p53 were detected in both mRNA and protein levels by mRNA Slotblot and Westernblot.The results of influence on expression of these genes by lithium were as follows:①In HL60,bcl2 and cmyc were downregulated in both mRNA and protein levels;②In U937,p53 and cmyc reduced in mRNA level;③As for A2780,the levels of p53 and cmyc protein and bcl2 mRNA decreased.The results suggest that these genes may play some role in the process of apoptosis induced by lithium.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1998年第1期5-11,共7页
Chinese Journal of Microbiology and Immunology
基金
中国医学科学院青年基金
关键词
锂
细胞凋亡
肿瘤细胞
Lithium\ Apoptosis\ Cell line Tumor cell,cultured
