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AIM To find a new method for purifying the active compound 5F isolated from Pteris semipinnata L.(PsL ) and observe its enhanced cytotoxicity when combined with other antitumor agents. METHODS Silica gel combined with AgNO 3 was made to purify 5F. Cytotoxicity was detected with trypan blue dye exclusion assay. RESULTS After purification, the concentration of compound 5F in purified products was higher than 99%. The inhibitory rates of 5F combined with 5 Fu or CDDP or VCR were higher than that of these drugs used alone. CONCLUSIONS Compound 5F could be purified effectively using silica gel combined with AgNO 3. 5F could enhance the cytotoxicity on HL 60 and K562 cells of the drugs mentioned above. The different effect of these agents on the various phases of cell cycle kinetics may explain the enhanced effect of 5F.
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OBJECTIVE：To investigate the use of micellar paper chromatography on the isolation and identification of flavonoids and their soluble derivatives.METHOD:Quercetin,Genistein and their sulphate derivatives were separated and identified by SDS solution above critical micelle concentration (CMC) on paper chromatography.The concentration of SDS,ratio of organic modifier and their concentration,fluorescence were investigated.RESULTS:The best concentration of SDS micelle was 0.01～0.02mol*L-1,and the spot effects of micellar chromatography could be improved by adding 2%～4% isopropanol (or n-amyl alcohol)into the mobile phase.CONCLUSION:The simple,rapid,accurate and qualitative method could be used to analyze and isolate flavonoids and their soluble derivatives.
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AIM: To investigate the changes of cytosolic free calcium concentration ([Ca 2+ ] i) and expression of Bcl-2 in HL-60 cells treated by 6F isolated from Pteris semipinnata L. (PSL), and to discuss the relations between calcium ion and cytotoxicity and DNA fragment induction effects of 6F. METHODS: HL-60 cells were used as in vitro model. [Ca 2+ ] i was measured on fluorescent spectrophotometry using Fura-2/AM as Ca 2+ indicator. Bcl-2 expressing level was measured by flow cytometry. Tetrazolium salt (MTT) and diphenylamine staining methods were applied for cytotoxicity assay and DNA fragmentation detection, respectively. RESULTS: [Ca 2+ ] i increased obviously in a dose and time dependent manner after treated HL-60 cells with 6F. 6F decreased the expressing level of Bcl-2. Adding 2 mmol/L Ca 2+ to the medium, or 1 mmol/L EDTA to chelate Ca 2+ , or 4 ?mol/L calcium ionophore A 23187 to increase the concentration of cytosolic Ca 2+ , the DNA fragment induction by 6F was not affected, whereas the cytotoxicity of 6F was enhanced. 250 ?mol/L Zn 2+ attenuated the DNA fragment induction, and the cytotoxicity of 6F against HL-60 cells was enhanced significantly. CONCLUSION: It was speculated that the decreased expressing of Bcl-2 by compound 6F was related to increased [Ca 2+ ] i in HL-60 cells, and DNA fragment induction was possibly catalyzed by Ca 2+ - independent DNase.
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Object To prepare the water soluble quercetin arginine complex (QAC) and widen the administration path of quercetin (QUE). Methods Definite QUE and L arginine were refluxed in alcohol to prepare QAC. The QAC structure was identified by micellar paper chromatography, UV spectrometry, IR spectrometry, and X ray diffraction. Results QAC was prepared from QUE and L arginine in molar ratio 1∶1. The inhibitory activity of QAC that existed stably in room temperature on cancer cell growth was as strong as that of QUE, and the solubility of QAC in water was remarkably enhanced. Conclusion The above preparation method is simple and available, and it is suitable to improve the bioavailability.
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Objective To study the effect of a diterpenoid 5F (ent-11?-hydroxy-15-oxo-kaur-16-en-19-oic acid) isolated from Pteris semipinnata on the expressions of NF-?B (P65), Smad3 protein and NF-?B (P65), ETS-1 mRNA, and to investigate the antitumor mechanisms of 5F. Methods MTT assay was used to examine the effect of 5F on proliferation of HO-8910PM cells. The expressions levels of NF-?B (P65) and Smad3 proteins were assessed by Western blot analysis. RT-PCR assay was used to assess the expression levels of NF-?B (P65) and ETS-1 mRNA. Results The proliferation of HO-8910PM cells can be inhibited by 5F in a dose-dependent manner. The expression levels of NF-?B (P65) and Smad3 proteins decreased obviously in HO-8910PM cells after treatment with 25—100 ?mol/L 5F for 24 h, and the effects appeared in a dose-dependent manner. 5F down-regulated significantly the expression of NF-?B (P65) mRNA, down-regulated gently the expression of ETS-1 mRNA. Conclusion The antitumor activity of 5F is involved in the expression of NF-?B (P65), Smad3 protein and NF-?B (P65), ETS-1 mRNA.
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AIM: To explore the effect of luteolin on the metastasis of ovarian carcinoma HO-8910PM cells in vitro, and to elucidate its mechanisms. METHODS: The in vitro invasion and mobility of HO-8910PM were measured by Boyden chamber assay after exposure to luteolin for 6 h. Gelatinase activity was tested by gelatin zymography assay. The expression of tissue inhibitor of metalloproteinase-1 (TIMP-1), nm23 mRNA and that of ERK2 protein were tested by RT-PCR and Western blotting, respectively. RESULTS: Luteolin significantly inhibited in vitro invasiveness and mobility in HO-8910PM cells in a dose dependent manner as measured by Boyden chamber assay. Luteolin inhibited the MMP-9 secretion from HO-8910PM cells. Luteolin did not affect the mRNA expression of TIMP-1 and nm23. Luteolin also decreased ERK2 expression. CONCLUSION: Luteolin dose-dependently inhibited the metastasis of HO-8910PM cells in vitro. Inhibition of MMP-9 secretion and ERK2 expression may be involved in the anti-metastasic process of luteolin.
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Aim To study the effect of resveratrol on the expression of FAK and the level of phosphorylated FAK, and to investigate its possible mechanism of resveratrol on the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells line. Methods MTT assay was used to examine cytotoxicity of resveratrol on HO-8910PM cells after 24 h treatment; The expression of FAK protein and the level of phosphorylated FAK were assessed by Western blotting analysis. Results MTT assay showed that resveratrol had weak cytotoxicity on HO-8910PM cells after 24 h treatment, its IC50 value was 163.40?2.48 ?mol?L-1. The expression of FAK and the level of phosphorylated FAK decreased obviously in HO-8910PM cells treated with 25~100 ?mol?L-1 resveratrol for 24 h. Conclusion The mechanism of resveratrol inhibiting the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells line involves the expression of FAK and the level of phosphorylated FAK.
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Aim To study the effect of cantharidin on the expression of NF-?B(P65), FAK protein and the level of phosphorylated FAK, and to investigate posssible mechanism of cantharidin on the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells. Methods MTT assay was used to examine the effect of cantharidin on proliferation of HO-8910PM cells after 24 h treatment; The expression levels of NF-?B(P65), FAK protein and the level of phosphorylated FAK were assessed by Western blot analysis.Results The expression level of of NF-?B(P65) protein decreased obviously in HO-8910PM cells treated with 5～20 ?mol?L -1 cantharidin for 24 h, and the effects appeared in a dose-dependent manner. Cantharidin significantly up-regulated the expression of FAK, down-regulated the level of phosphorylated FAK.Conclusion The antitumor activity of cantharidin is involved in the expression of NF-?B(P65) and the level of phosphorylated FAK.
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Aim To investigate the inhibitory effect and its posssible mechanism of 5F from Pteris semipinnata L on the metastasis-associated ability of human highly metastatic ovarian carcinoma HO-8910PM cells in vitro.Methods MTT assay was used to examine the effect of 5F on proliferation of HO-8910PM cells after 24 hours treatment; Transwell Chamber assay was performed to determine the effect of 5F on invasion and migratory capacity of the cells; Effect on adhesion potential of HO-8910PM cells was tested by cell-Matrigel adhesion assay; The expression levels of NF-?B(P65) and VEGF proteins were assessed by Western blot analysis.Results 5F significantly inhibited invasion, migration, and adhesion capacity of HO-8910PM cells in vitro, their inhibitory rates after treatment with 50 ?mol?L -1 5F for 6 h were (37.57?0.62)%, (28.42?0.67)%, (46.07?4.49)%. 5F d istinctly down-regulated the expressions of NF-?B(P65) and VEGF protein. Conclusion 5F can inhibit the migration, invasion and adhesion of HO-8910PM cells in vitro,Its possible mechanism may be involved in the reduction of the expression level of NF-?B(P65) and VEGF. 5F might be potential drugs to inhibit tumor metastasis.