Purification And Characterization Of Intracellular Protein PSPB-2 From Pholiota Squarrosa AS5.245

At present,cancers have still been one of the important factors in human death.The main treatment mcthodologies to tumors include operation,chemotherapy,radiotherapy, which in one side bring hopes and on the other side induce various injuries to patients.And the side effects of chemotherapy,radiotherapy are much bigger to postpone the recovery. Therefore high attention has been paid recently in thefields of biology and medine to select new drugs,especially natural medicines with higher efficiency,lower toxicity and low cost.Pholiota squarrosa is mycelium of fungi,which belongs to Basidiom ycotina, Hymenomycete,Agaricales,Strophariaceae,Pholiota(Fr.) kumm.At present there is no research in the antitumor activity of Pholiota squarrosa.In this study water-soluble proteins from Pholiota squarrosa by femtation have been isolated and its biological activities have been studied.1:According to the Pyriculariaoryzae bioactivity method,We build Rhizoctonia solani bioactivity method.The activity substance of Pholiota squarrosa have been studied guided by Rhizoctonia solani bioactivity method.2:The proteins was purified by ammonium sulfate precipitation,ion exchange chromatography and gel filtration,SDS-PAGE electrophoresis of the purified fraction mitigated a simple band,The molecular weight of the enzyme was 38.28KDa The purified protein have markable antiproliferative activity.In vitro,the antiproliferative activity toward tumor cells A549 cells induced by Pholiota squarrosa proteins were assayied with antiproliferative method,when contains 20μg/ mL proteins the growth inhibition was higher than 65%.3:The study shows that the proteins purified from Pholiota squarrosa can induce cancer cell apoptosis.We used JC-1 for in-situ detection of mitochondrial membrane transition events in live cells,which provides an early indication of the initiation of cellular apoptosis.The collapse in the electrochemical gradient across the mitochondrial membrane,we found the proteins can induce collapse of mitochondrial.Futher more we use the TUNEL kit,it's able to see the Fluorometric of break DNA 3'-OH. 4:Endonuclease activity had been found toward plasmid PUC18.