Experimental study on Qinghuang Powder through inhibition of BCL-2,XIAP-1,c-IAP synergetic induced apoptosis in KG1a cells
WU Jian-wei1 HUANG Jian-xu1 ZHENG Rong1 YE Yong-bin2 FAN Jia-xin1 LI Yuan-ming1 ZENG Yingjian3▲ GUO Kun-yuan4
1.Department of Hematology,Jiangmen Hosptial of Traditional Chinese Medicine Affiliated to Jinan University,Guangdong Province,Jiangmen 529000,China;2.Department of Hemoendocrinology,People′s Hosptialof Zhongshan City,Guangdong Province,Zhongshan 528400,China;3.Department of Hematology,Hosptial Affiliated to Jiangxi University of Traditional Chinese Medicine,Jiangxi Province,Nanchang 330000,China;4.Department of Hematology,Zhujiang Hosptial,Southern Medical University,Guangdong Province,Guangzhou 510282,China
Abstract：ObjectiveTo study the effects of Qinghuang Powder(composed of two components of Indigo naturalis and Realgar),which the active ingredients are Indirubin and Arsenic Disulfide(As2S2),alone and in combination with cytotoxic effect on non M3 AML KG1a cells,and to investigate the mechanism of cytotoxic effect.MethodsThe cell morphology of KG1a cells acted on with Indirubin and As2S2was observed by Wright Gimsa staining;the rate of cell apoptosis of KG1a cells acted on with Indirubin and As2S2was detected by flow cytometry;CCK-8 detected for apoptosis rate of KG1a cells acted on with Indirubin and As2S2;the expression of Bcl-2,Smac,XIAP-1,c-IAP,Caspase-3 protein of KG1a cells was detected by Western blot.ResultsBoth Indirubin and As2S2had inhibitory effect on proliferation of KG1a cells.Compare with single dose,the combination of Indirubin and As2S2had a higher cell proliferation inhibition rate and apoptosis rate(P＜0.05).When Indirubin and As2S2combined with 1∶1,the combined index was less than 0.9 through CompuSyn,which could also influenced the apoptosis highly,increased the expression of Caspase-3 and Smac protein,and inhibit the expression of BCL-2,XIAP-1 and c-IAP protein.ConclusionThe Indirubin and As2S2,the ingredient of Qinghuang Powder,can inhibit the proliferation of KG1a cells and induce its apoptosis,the mechanism may be related to inhibition of the expression of BCL-2,XIAP-1,c-IAP protein,promote the expression of Caspase-3,Smac protein.Qinghuang powder can be used for the treatment of AML of conventional drug resistance and provides the relevant experimental basis for the treatment of non-M3 type AML.
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