Clinical Observation Of Qilianyisui Granule Combined Chemotherapy In The Treatment Of Acute Leukemia And Its Effect On NB4 Telomere-binding Protein In Leukemia Cell Line

Objective:The purpose of this study is to observe the clinical improvement of Qilianyisuiqingdu Granule(QG)combined with chemotherapy in treatment of acute leukemia,evaluate QG combined with chemotherapy efficacy and safety of acute leukemia,investigated the possible mechanism of quercetin on the proliferation and apoptosis of NB4 cells by testing the expression of three telomere-binding protein,to seek better experimental support,in order to provide new ideas and methods to improve the clinical efficacy of leukemia.Methods:1.Clinical test: Choose 30 patients with acute leukemia from department of Integrated TCM &Western Medicine and hematopathy in Gansuprovince tumor hospital in September 2013 to December 2015.According to the visiting sequence were randomly divided into two group,treatment group(Chinese medicine combined with chemotherapy)were 15 cases,control group(chemotherapy alone)were 15 cases.Control group were treated by chemotherapy and symptomatic treatment,treatment group take QG(3 bags each time,three times a day)on the basis of control group.After 2 course of chemotherapy,to compare the clinical efficacy,total score and peripheral blood,toxic side effects and complications of two groups of patients.2.In vitro:(1)Cultured the human acute single nuclear leukemia NB4 cells.(2)Cellular viability was measured by CCK-8 assay,and the cells were incubated in the absence or presence of increasing concentration of QG fordifferent time,which determined the IC50 of the experimental group.(3)The protein expression levels of each group of RAP1?TRF1?TRF2?TZAP were assessed by Western-blot analysis.(4)The levels of RAP1?TRF1?TRF2?TZAP m RNA were detected by real-time fluorescent quantitative PCR(RT-PCR)analysis.Results: 1.Clinical trials:The total score and improvement rate of TCM symptoms comparison:after treatment,was significantly lower thanthat of control group,the difference was statistically significant(P<0.05);Hemogram comparison: the change of WBC,RBC,Hb,PLT of two groups after treatment display that the hemogram recovery degree of treatment group was significantlybetter than that of control group,the difference was statistically significant(P<0.05).2.In vitro:(1)CCK-8 assay showed that inhibited cell growth and decreased cell viability in a dose-and timedependent manner,the experience group of IC50 was 62?g/m L.Therefore,in our study,we choose with the medium concentration 60?g/m L(lower than IC50)at 48 h.(2)As shown by western blot analysis,as epicted showed that compared with controls,the experimental group elevated TRF1 protein level and degraded TRF2?TZAP?RAP1 protein level(P<0.01),there were significant differences.In addition,the findings demonstrated that TZAP and TRF1 were positively correlated(r=0.922),TRF2 and TRF1,TZAP were negatively correlated(r1=-0.863?r2=-0.843).Conclusion:QG has a synergistic and attenuated effect on chemotherapy,and is effective and safe in the clinical adjuvant treatment of acute leukemia.In vitro experimental study of QG showed a certain degree of anticancer activity,and its mechanism may be related to elevated TRF1?TZAP?RAP1 protein level and degraded TRF2 protein level in order to induce the apoptosis of leukemia cells,and provide experimental theoretical basis for the clinical application of QG.