The Effect Of Cucurmosin Against Chronic Myeloid Leukemia Cell K562

Objective: The aim of this study is observation the inhibiting effect in human Chronic Myelogenous Leukemia cell line (K562) by Cucurmosin(CUS). Estimate the anti-tumor effect of CUS in vivo with Human Chronic Myelogenous Leukemia cell in suit and xenograft in NOD/SCID mice model. To provide some potential theoretical foundation on anti-CML of Cucurmosin.Methods: Compared with TCS, detect the inhibiting effect in human Chronic Myelogenous Leukemia cell line (K562) by Cucurmosin(CUS)with MTT. K562 cells were injected subcutaneously into the severe combined immunodefiency (SCID) mice repeatedly to establish the Tumor transplanted subcutaneously model in the NOD/SCID mice.The human leukemia mice model was established by implanting K562 cells into the caudal vein of SCID mice . Peripheral blood,bone marrow with Wright staining, myeloid antigens( CD13,CD33,CD117) of bone marrow cell with flow cytometry, RT-PCR detect bcr/abl fusion gene of bone marrow and histipathology were used to identify model. Judging the therapeutic effect of CUS in in vivio .Results:Cucurmosin inhibited proliferation of K562 cells in a dose-dependent manner, its efficacy was stronger than that of TCS.Their IC50 of Cucurmosin for 24h,48h and 72h were 8.59 mg/L,3.85 mg/L and 2.39mg/L respectively. K562 cells were injected subcutaneously into the severe combined immunodefiency (SCID) mice repeatedly to establish the Tumor transplanted subcutaneously model in the NOD/SCID mice. The mouse were in awful condition and weight loss obvious after model building in 5 weeks; Part of animals showed tumor block, for example eyes,crus and ovarian etc;Numeration of leukocyte of Blut-Routine-Untersuchung brought into correspondence with Peripheral under the microscope.PWBC was very low after irradiation ,WBC were seen rarely. So WBC is not proportional to the day of model building , Mature granulocyte mainly in PB and observed the immature leukemia cells in peripheral hemogram ,rarely; Peripheral Blood NAP stain integral was very low ;Bone marrow cell with Wright Stain under the low-magnification microscope showed immature cells visible easily; Dynamic viewing(after model building in 2,4,6 and 8weeks ) myeloid antigens( CD13,CD33,CD117) of bone marrow cell with flow cytometry ,report showed the blank group(irradiate without inject k562 cells)were not higher and mutative, on the contrary negative group (irradiate with inject k562 cells)myeloid antigens increase gradually and proportional to the days of model building;Ngative group is near death ,which bone marrow cell detect CD13,CD13,CD117 with FCM, meanwhile blank group did it. The two groups were statistically significant;The CD33 of liver and spleen with flow cytometry showed no difference between blank group and negative group. The total positive rate of CD33 expression was low and no change in them; Ovaries of female mouse were infringed with cancer. Organs infringed with cancer,liver,spleen,lung and kidneys did pathology.Organs infringed with cancer showed typical tumor tissue forms,the others did not see cancer infringed and obviously abnormal; RT-PCR detect bcr/abl fusion gene, bone marrow ,tumour,and K562 cells can be detected bcr/abl fusion gene,the others(liver,spleen and blood) not see.In vivo, the tumor weight inhibitory rate of Cucurmosin at 1,0.5,0.25 mg/kg was 59.43%,53.45% ,19.18% ,respectively;blood model, life extension rate of Cucurmosin at 1,0.5,0.25 mg/kg was 16.1%,36% ,24.9% ,respectively. Conclusion:1 Cucurmosin inhibited proliferation of K562 cells in a concentration- and time- dependent manner..2 .The method of qualification test of CML blood model of NOD/SCID with K562 , which give first place to bone marrow slides, myeloid antigens( CD13,CD33,CD117) of bone marrow cell with flow cytometry, RT-PCR detect bcr/abl fusion gene of bone marrow.3 Cucurmosin can significantly inhibit the growth of human CML cell line K562 in NOD-SCID mice.