The Regulation Effect Of Notch And Correlated Signa Conduction System To Cell Proliferatiuon And Differation

Notch signal conduction is a highly conservative signal conduction.It plaies critical affection in wide field of cell differentiation and development process,especially in determining the cell differentiation fate.There are a aroup of possessing self-renewal potential stem cells in leukemia----leukemia stem cells(LSC).The existence of LSCs become an important factor of leukemia relapse and refractory leukemia.The role of Notch pathway must coordination with other signal pathway to play.The activation of Notch pathway through the phosphatidylinositol 3-kinase(PI3K)—Akt pathway to promote proliferation.The mammalian target of rapamycin (mTOR) was shown to be a key kinase acting downstreamof the activation of the PI3K to regulate cell proliferation.Notch,PI3K—Akt,mTOR pathway play critical regulative function in stem cells' self-renew, proliferation,differentiation and so on.【Objective】1,The affection of Notch,PI3K—Akt,mTOR pathway inhibitor and combinateapplication to leukemia stem/progenitor cells' clonal formation ablility,self-renew capability,proliferation,differentiation level, apoptosis level,cell generation cycle,and LSC quatility.2,To investigate the regulatory mechanism of LSCs' self-renew,proliferation,and differentiation capability.3,To discovery the best combination schedule to inhibit leukemia cells' proliferation,clear LSC and promote HSCs' self-renew capability.【Methods】1,The fresh bone marrow from preliminary diagnosis leukemia patients were isolated using Ficoll solution and then had been cultured with DAPT and/or rapamycin and/or Ly294002.2,To detection the express level of cell surface CD34;CDS8;CD123; CD117;CDIIb;CD14 by flow cytometry.3,To detection the apoptosis level and cell life cycle by flow cytometry.【Results】1,DAPT,rapamycin and Ly294002 can inhibit the cell proliferation,clonal formation,self-renew capability(P＜0.05).The combination schedule of DAPT with Ly294002 is better than DAPT in inhibiting self-renew.2,DAPT or rapamycin can decrease CD34~+ cells' quantity.DAPT,rapamycin and Ly294002 all can decrease CD117~+ cells' quantity.The combination schedule of DAPT with rapamycin is better than DAPT or rapamycin along.3,No find any evidence of DAPT,rapamycin and Ly294002 can induce leukemia cells to differentiate to mononuclear cells.4,DAPT can decrease the quantity of CD34~+D38~-,CD123~+ cells.The combination schedule of DAPT with Ly294002 is better than DAPT alone.5,The combination schedule of DAPT with Ly294002 can induce apoptosis. 6,The combination schedule of DAPT with Ly294002 can decrease the quantity of S stage cells.【Conclusion】1,DAPT,rapamycin and Ly294002 can regulate the cell proliferation,differentiation,elonal formation,self-renew capability,apoptosis and cell life cycle level.2,DAPT especially the combination schedules of DAPT with Ly294002 or rapamycin are better than only one inhibitor schedule.