摘要
目的:研究新型抗肿瘤药物6'-羟基爵床素A(6'-hydroxy justicidin A,JR6)对HepG2细胞的内质网应激及胞内钙离子浓度变化的影响及其意义。方法:HepG2细胞经过终浓度分别为0,0.615,2.45,9.8μmol.L-1的JR6处理24h后采用实时定量RT-PCR技术、蛋白印迹(Western blot)法检测细胞中X-盒结合蛋白1(XBP-1)mRNA,IRE1α(inositol-requiring enzyme-1α),GRP78/BiP(glucose regulated protein/BiP),及Bcl-2(B cell leukemia 2)蛋白表达量的变化。激光扫描共聚焦显微镜测JR6作用24h后细胞内静息钙浓度变化及IP3R钙泵抑制剂光溜海绵素C干预后JR6的作用变化。结果:IRE1,XBP-1mRNA,Bip,Bcl-2蛋白的表达量均与JR6剂量负相关。高剂量组的各蛋白表达量与空白组相比,均下降了40%左右。共聚焦显微镜观察结果显示JR6引起细胞内钙离子浓度升高了2倍左右;加入IP3R钙泵抑制剂后,JR6不能引起细胞内钙升高。结论:JR6通过内质网钙泵IP3R升高细胞内的钙浓度,破坏细胞内的钙稳态;同时引起了内质网应激,并抑制了未折叠蛋白反应的相关因子。
Objective： To research the effect of 6-hydroxy justicidin A（JR6） on endoplasmic reticulum stress（ERS） and Ca2 ＋ in HepG2 cells.Method： Semiquantitative RT-PCR and Western blot were used to investigate the expression levels of X-box binding protein 1（XBP-1） mRNA,Inositol-requiring enzyme-1α（IRE1α）,Glucose regulated protein/BiP（GRP78/BiP）,B cell leukemia 2（Bcl-2） proteins in HepG2 cells which were treated with 0,0.615,2.45,9.8 μmol.L-1 JR6 separately for 24 h.Laser confocal scanning microscopy was used to study the difference of intracellular calcium between medicine group and IP3R（inositol 1,4,5-trisphos-phate receptor） inhibited group.Result： The expression of IER1,GRP78 / BiP Bip,Bcl-2 proteins and XBP-1 mRNA have a reduction in approximately 40% with high JR6 group.It is found that the JR6 could elevate the intracellular calciumion concentration more than two times,and this effect was disappeared with the IP3R inhibitor（Xestospongin C）.Conclusion： JR6 through IP3R way elevates intracellular free Ca2 ＋ concentration,and arouse ERS.At the same time,JR6 can inhibit elements which related to unfolded protein response（UPR）.
出处
《中国实验方剂学杂志》
CAS
北大核心
2013年第13期212-215,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(30873090)
关键词
HEPG2
肿瘤
内质网应激
钙稳态
未折叠蛋白反应
HepG2
tumor
endoplasmic reticulum stress
intracellular free Ca2 ＋ concentration
unfolded protein response
