| Objective : The current treatment for leukemia substantially rely on chemotherapy . Most drugs sued in chemotherapy have serious side effects , which include the suppression of bone marrow's hemopoetic function and the suppression of human body's immunological function and these side effects bring about many undeniable complications such as gravious infection , bleeding , and so on . As a result , the life qualities of the patients who are diagnosed as leukemia are badly affected , even their dear lives are severely endangered .Since 1980s' , researchers all over the world have discovered some natural medicine which have only minor side effects and at the same time have many advantages including the emphasis on integration and the consideration for both action and compensation . the effective component extracted from traditional Chinese medicine and natural medicine has become another important ami -leukemia option . In recent years , Chinese researchers have begun to experiment on traditional Chinese medicine's effect in vitro of inducing tumor cell apoptosis and differentiation . Until now , many effective components extracted from traditional Chinese medicine have been found to induce tumor cell's apoptosis and differentiation . Kangbailing is an improved agent based on Ginseng-largehead atractylodes rhizome mixture , which is gabricated in our hematology department for many years . the objective of our study is to give the experimental evidence for a high efficiency-low toxicitytraditional Chinese medicine which poses the characteristic of selectively kill the leukemia cells . To examine KBL's functional mechanism at both cellular level and molecular level , the in vitro experiment is designed to test whether KBL can kill leukemia cells , induce apoptosis and differentiation of leukemia cells .Methods: 1 . MTT assay and trypan blue resisting staining method are used to find out KBL's inhibition on HL-60 (a human leukemia cell line) .2 . Morphological analysis , flow cytometry (FCM) assay and electrophoresis assay are used to examine cell apoptosis .3 . Using retinoic acid as the positive control , morphological analysis , NET reduction assay and ink phagocytizing assay are used to examine HL-60' differentiation after treating HL-60 with KBL (6.25 mg/ml) for 72 hours4 . 2-step method is used to cloning culture primary leukemia progenitor cells and normal progenitorcells and observe different KBL concentrations' effect on HL-60's cloning formation.Results :1 . HL-60 cells' proliferation was inhibited by KBL concentrations from 6.25 mg/ml to 50 mg/ml .This effect shows dosage and time-dependent relation . IC50 is 25.0 mg/ml. 2 . Typical morphological changes were found in morphological analysis ; the sub-G1 peak was found in FCM analysis and the typical ladder was found in DNA agarose gel electrophoresis analysis . These results showed KBL's definite effect on HL-60's apoptosis . 3 . In NBT reduction assay and ink phagocytizing assay , after treating HL-60 cells with KBL (6.25 mg/ml) for 72 hours . The NBT positive reacting rate and in phagocytizing rate were 50.8% and 56.0% respectively , which were much higher than the negative control group , and statistical analysis showed significant difference between the two groups .The rates of the two assays in the retinotic acid group (the positive control group) is 55.3% and 69.0% , which were slightly higher than the KBL group , but statistical analysis showed no significant difference between them . 4 . In the cell culture analysis , KBL can markedly inhibit leukemia progenitor cell cloning formation but it couldn't inhibit normal progenitor cell cloning formation. Statistical analysis showed significant difference between these two groups .Conclusion : KBL can inhibit HL-60's proliferation and this effect shows dosage , time-dependent relation . KBL can induce apoptosis and it can interfere the cell-cycle phase in HL-60 and cause cell remain in d phase . KBL can induce HL-60's differentiation , and the effect is similar with retinoic acid . KB...
|
PDF Full Text Request