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The Inducement and Differentiation of Chinese Medicine and New Chemical Compounds to Cells from Blood

Author: GaoXia
Tutor: LiXingYu
School: Shanghai Normal University
Course: Zoology
Keywords: BPA-4 cell differentiation Caesalpinia Sappan L. Liposome Leukemia Cell Line K562 Cell apoptosis
CLC: R285.5
Type: Master's thesis
Year: 2008
Downloads: 93
Quote: 1
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Abstract


Cancer has been the second killer to people,because of the industrial pollution and living habits of human beings.To find out low-toxicity,high-effective anti-cancer medicines,and to apply accurate data and methods for cancer-treating and helping treatment;we took the Chinese medicine Caesalpinia Sappan L（CSL）and new chemical compounds as targets,did research on their inducement and regulation to cancer cells and stem cells like CD117 from cord blood,screening out the effective medicines and effective concentration like IC50,and further tried to explain the reasons.Applying cell culture technology,molecular biology technology,nano-technology and immuno-technology,using fluorescent microscopy,optical microscopy,laser scanning confocal microscopy（LSCM）,scanning electronic microscopy（SEM）,transmitting electronic microscopy（TEM）,PCR and other research machines,we did researches on the positive regulation of Caesalpinia Sappan L（CSL）to the stem cells（like CD117+）and the negative-regulation of CSL to the K562 cells（leukemia cell line）,also using limposome capsulated the water solution of CSL to improve the medicine-delivery;In the mean time,we also screened about 200 new chemical compounds to find out effective medicines and did further research on their negative-regulation of CSL to the K562 cells.Results:1.the water solution of CSL inhibited the reproduction of K562 dramatically （p＜0.05）,and the concentration and the inhibition rate（IR）was correlated(r24h=0.90001004、r48h=0.984114411、r72h=0.973074678),with the climax at 48h 100μg/ml,25μg/ml of CSL had the same effect with 5-Fu（50μg/ml）;the CSL liposome could decrease the clone-forming (P＜0.05,r=0.978,IC50=129.92μg/ml);2.the CSL liposome induced cell apoptosis:the apoptosis modality like a decrease in cell volume,nuclear debris,apoptotic bodies and etc could be seen under the fluorescent microscopy,optical microscopy,LSCM and TEM,and the experimental group was apparently different from the control group（P＜0.05,0.9＜r＜1）; DNA ladder showed on the agarose gel electrophoresis and sub-double body showed on the FACS;3.genes associated with apoptosis like p53、p16、caspase-3、bcl-2、Rb were amplified by RT-PCR;4.the radius of the self-made immuno-fluorescence nanoparticles（IFN）was about 122nm under the AFM and SEM,and had no evident inhibition to the K562,Hela and mice primary bone cells（p＞0.05）;5.the solution of CSL could induce human cord blood stem cells differentiate to CD33+:the best concentration is 25μg/ml at 7d,50μg/ml at 2.5d and 3.5d with r2.5d=0.9663、r3.5d=0.715526;6.the solution of CSL could induce human cord blood stem cells differentiate to CD19+:the positive rate was over 70%at 2.5d and 3.5d, totally different from the Co.group（＜0.05）;when the cultured time was prolonged,the positive rate was even with the climax 30%at 50μg/ml;7.the solution of CSL induced human cord blood stem cells differentiate to CD4+:the positive rate was about 88%and 92% at 2.5d and 3.5d respectively,the correlation is evident,r2.5d=0.9818665,r3.5d=0.8355519;8. the solution of CSL could introduce CD117 stem cells differentiate to CD4+ mature cells with r=0.910386 and 37.5μg/ml as the best concentration;9.54 new chemical compounds,which inhibited the cell reproduction dramatically（p＜0.05）,were screened out from 200 compounds by MTT,and the IR and the concentration was correlated（0.9＜r＜1）;10.BPA-4 induced cell apoptosis,the modality like chromatin condensation,nuclear debris,apoptotic bodies and etc showed under the fluorescent microscopy,optical microscopy,ladder showed on the DNA agarose gel electrophoresis and fivefold sub-double body compared to the contrast showed on the FACS（18.6%:3.1%）,besides,cells were stayed at G0-G1 phase by FACS.Conclusion:1.the solution of CSL can inhibit the reproduction of K562 and introduce cell apoptosis,and maybe carry out this function through control the expression of genes like p53、bcl-2、p16、Caspase-3 and Rb.2.the solution of CSL have the positive-regulation function to introduce human cord stem cells differentiate to granule-single progenitor cells and mature immuno-cells.3.54 kinds of the new chemical compounds among 200 were screened out with obvious inhibition to the reproduction of K562,among them BPA-4 could induce K562 cell apotosis and cause the cells stayed at the G0-G1 phase.

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CLC: > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology
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