Effects Of Berberine Hydrochloride On The Apoptosis And Expression Of Survivin Gene In Jurkat Cells

Acute lymphoblastic leukemia (ALL) is a malignant tumor derived from a single B or T lymphocyte precursor cells.ALL was common in children and adolescent,which had a better prognosis in children.In adults,the use of standard chemotherapy could often achieve the first remission of ALL,but the recurrence rate was as high as70-80%.Traditional chemotherapy drugs had toxic effects on normal tissues,especially the hematopoietic system,which limited the dose of chemotherapy drugs,so seeking efficiency and low toxicity treatment became the focus of current research.Berberine can be extracted from the Chinese medicine Coptis, Phellodendron, berberidis and other genus Berberis plants.The modern research shows that berberine had anti-tumor activity against a variety of solid tumors and hematological malignancies.The mechanism are related to induce tumor cell apoptosis,cell cycle arrest and inhibition of angiogenesis,but it has tolerable side effect to normal tissue.Our research was carried out to investigate whether berberine hydrochloride could affect the proliferation,apoptosis,cell cycle and expression of Survivin gene in Jurkat cells.IC50value and cytotoxity of Jurkat cells were detected by modified MTT assay,nuclear changes by fluorescent staining with Hoechst33342;cell apoptotic rate,by FCM(Flow cytometry) with Annexin-V-FITC+PI staining assay;cell cycle,by FCM(Flow cytometry) with PI staining assay; expression of Survivin gene,by Semi-quantitative RT-PCR.The result shows that the cell viabilites were significantly inhibited by berberine hydrochloride at the concentration of0-10μg/ml.The cell survival rate was decreased with the increasing concentration of berberine hydrochloride. Furthermore,berberine hydrochloride inhibited cell growth in a time dependent manner.If treated at the same concentration of berberine hydrochloride,fewer cells would survive when the cells were treated for an increasing time.The24h,48h,72h IC50of berberine hydrochloride on Jurkat cell were6.94μg/ml,3.94μg/ml,2.45μg/ml. After treated by0μg/ml,5μg/ml berberine hydrochloride for48h,Jurkat cells observed by fluorescent staining with Hoechst33342,showed a typical apoptotic morphology,nuclear deformation,nuclear chromatin condensation and apoptotic bodies. After treated by0,0.1,0.5,1,5,10μg/ml berberine hydrochloride for48h,the Jurkat cells in early apoptosis were2.06%±0.51%,3.60%±1.05%,3.38%±1.37%,3.97%±1.53%,12.77%±1.07%,14.27%±2.11%.The results of PI staining assay show that the cell cycle was arrested in the S phase,Furthermore, the expression of Survivin gene decreased as measured by RT-PCR in Jurkat cells treated by berberine hydrochloride.In conclusion,berberine hydrochloride can inhibit Jurkat cells proliferation,induce cells apoptosis and cell cycle arrest,which may through down-regulated the expression of Survivin gene.