The Effect And Mechanism Of Tanshinones On Hematological Malignant Tumor

Objectives: Based on anti-tumor effect of Tanshinones compounds,this study mainly detected the function of Tanshinones compounds(dihydrotanshinone?,tanshinone ?A,cryptotanshinone,tanshinone?,)on T cell lymphoma cell lines(Jurkat),observing inhibition of proliferation,cycle arrest andinduction of apoptosis.On that base,exploring the inhibition,induction of apoptosis and cycle arrest of Tanshinones compounds to Hematological malignant tumor cells(multiple myeloma cell lines U266,myeloid leukemia cell lines NB4 and K562,T cell lymphoma cell lines Jurkat)mediating by NF-?B,MAPK and PI3 K three signaling pathways.Methods:(1)Culturing Jurkat cell in vitro,According to the concentrat-ion of Tanshinones compounds(1.25,2.5,5,10,20,30?mol/L),Jurkat were divided into six groups for each of these drugs.It was designed with daunorubicin(0.7,1.4,2.8,5.6,11.2,16.8?mol/L)which own the considerable quality as positive control group.At the same time,it was designed a negative control.After cultured for 24,48,72 hours,the inhibition of the groups were detected by CCK8 method.Jurkat were treated by 20?mol/LTanshinones for24 hours.Then,PI single staining method was used to detect cell cycle arrest of the drugs(20?mol/L)on Jurkat cell line.Annexin V/PI double staining method was used to detect induction of apoptosis of the drugs(20?mol/L)on Jurkat cell line.(2)Culturing Hematological malignant tumor cell lines(NB4,K562,U266,Jurkat)which were collected After cultured for 24 hours with20?mol/L Tanshinones compounds stimulated.Extracted total protein,andstored at-80? for later use,then the concentration of the protein was detected meeting the requirements.Finaly,Western blot method was used respectively to detect the protein expression of NF-?B,MAPK and PI3 K three signaling for four kinds of cells.Results:1.Dihydrotanshinone?,Tan IIA,CPT and Tan I could inhibit proliferation of Jurkat.The effects of drugs depended on the time and concentration.Four Tanshinones compoundsstimulated Jurkat cell line: 24 hours,IC50 respectively were 16.40,12.53,49.47,54.19?mol/L;48 hours,IC50 respectively were 8.73,7.28,38.89,15.66?mol/L;72 hours,IC50 respectively were 2.95,2.65,17.43,10.28?mol/L.2.Jurkat cell line was treated by Tanshinones(20?mol/L)for 24 hours.Then,observing the cell cycle arrest by Flow cytometry.Compared with blank control cell group,in addition to Tan IIA,G0/G1 phase cells increased significantly(P<0.01);S phase and G2/M phase cells decreased,the S phase cells proportion of the group of CPT and the blank control group were no significant difference;The effect of the group of dihydrotanshinone?on Jurkat cell line was the most strongest.S phase cells of the group of the Tan IIA increased significantly,G0/G1 phase and G2/M phase cells decreased.3.Jurkat cell line were treated by tanshinones(20?mol/L)for 24 hours.Observing the induction of apoptosis by Flow cytometry.All the early apoptosis cells rate,the late apoptosis and necrosis cell rate increased significantly in the group of Dihydrotanshinone?which was Compared with the blank blank control cell group,Tan IIA,CPT and Tan I acting on Jurkat cell line(P<0.01).The effect of the group of dihydrotanshinone?on Jurkat cell line was the most remarkable,following respectively were Tan IIA >TanI>CPT.The effect of the group of CPT was the weakest.4.By using western blot method respectively,the protein expression of NF-?B,MAPK and PI3 K three signaling for four kinds of cells(NB4?K562?U266 ? Jurkat)were detected after cultured for 24 hours with 20?mol/L Tanshinones compounds stimulated.The result indicated that Tanshinones compounds significantly inhibited NF-?B and PI3 K two signaling pathways of Hematological malignant tumor cells line,but no effect for MAPK signal pathway.Conclusion:(1)Tanshinones compounds could inhibit proliferation of Jurkat cell line.The effects of drugs depended on the concentration and time.(2)Dihydrotanshinone?which own the strongest proliferation inhibition effect compared with daunorubicin which own the considerable quality,roling respectively after 24 hours,48 hours,the proliferation inhibition effect of Dihydrotanshinone ? to Jurkat cell line was significantly weaker than daunorubicin;72 hours later,the proliferation inhibition effect to Jurkat cell line Of the two drugs had no difference.(3)Tanshinones compounds on Jurkat cell lines,had the effects of cycle arrest and induction of apoptosis,the strongest one was the dihydrotanshinone?.(4)Tanshinones compounds could affect the cycle distribution of Jurkat cell line,the group of dihydrotanshinone??CPT and Tan I were blocked in G0 / G1 phase cell,the group of Tan IIA was blocked in S phase cell.(5)Tanshinones compounds acting on the Hematological malignant tumor cell lines(K562,NB4,U266,Jurkat),could significantly inhibit the NF-?B,PI3 K two signaling pathways.What,s more,mediating proliferation inhibition,cycle arrest and induction of apoptosis of Hematological malignant tumor cell.However,having no significant effect to MAPK signal pathway.