A Experimental Study On The Antineoplastic Activity And Induce-cell Apoptosis Activity Of Cortex Periplocae Extracts

Objective: To study the antineoplastic activity,induce-cellapoptosis activity and its mechanisms of Chinese medicinal herbCPE. To produce the scientific evidence for developing andmanufacturing new antitumor drugs.Methods: 1. The suppressive effect of CPE on theproliferation of BGC-823,K562,TE-13,MCF-7,SMMC-7721,BEL-7402,Hela and H22,colon26,S180 tumor cells wereinvestigated in vitro by using MTT colorimetric method andcolon forming method.2. The mouse modles bearing tumor were constructed byinjecting digitoplantarly colon26 tumor cells and injectingsubcutaneously S180 tumor cells inspectively, then were treatedwith CPE to observe the weight of tumor and the life span of themice to evaluate its antitumor activity in vivo.3. The apoptosis of human gastric carcinma cell BGC-823induced by CPE was studied by Gimsa staining, flow-cytometry(FCM), electric microscope and DNA agarose gel electrophoresistechnique. The mRNA and protein expression of bcl-2,bax andsurvivin were investigated by using semi-quantity RT-PCR andimmunocytochemistry staining technique, and the mechanism ofapoptosis was analysised.Results: 1. CPE inhibited markedly the proliferation oftumor cells such as BGC-823 ,K562 ,TE-13 ,MCF-7 ,SMMC-7721,BEL-7402,Hela cells from human ,H22,colon26and S180 cellsfrom mouse, with dose-dependent relationship. Forhuman gastric carcinma cell BGC-823,human erythroblest cellK562 and human esophageal carcinoma cell TE-13, the CPE hasa more inhibitive activity. The IC₅₀ for the three kinds of cell ofCPE are all less than 20μg/ml.2. After treatment with CPE, the general condition of testmice bearing tumor was much better than that of control micewith tumor growing slowly and life span being prolonged.3. After treatment with CPE, the BGC-823 cells presentedsome typical morphologic features and super-microstructuralchanges of apoptosis, including cell shrinkage, cytoplasmconcentration, nclius forming outwardly acute angle promineney,chromatin concentrating on karyotheca or forming meniscus,nuclear condensation, electric density increasing, nuclearfragmentation and formation of apoptotic bodies. Some typicalsubdiploid peaks before G₀/G₁ phase of BGC-823 cell induced byCPE were observed. The apoptotic rate of BGC-823 was 18.9%after 250μg/ml CPE treating for 48h. DNA agarose gelelectrophoresis showed characteristic "DNA ladder"pattern.After treatment with 250μg/ml CPE, the cells in G2/M phasewere blocked and increased remarkably. During the succedentresearch, used Semi-RT-PCR and immunocytochemistry staining...