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Objective To investigate the inhibitory effects ofPolyphyllin D (Paris saponin [PS]Ⅰ) and formosanin C (PS Ⅱ) on human glioblastoma U251 cells and its mechanism.Methods Glioma cell line U251 was cultured in vitro.Different treatments (PSⅠ and PS Ⅱ) were added into the medium cultured human malignant glioma U251 cells; and blank control group was also established.The effects of PSⅠ and PS Ⅱ on proliferation of glioma cells U251 in vitro was examined using methyl thiazolyl tetrazolium (MTT) assay.The nuclear morphology changes of apoptotic cells were detected by Hoechst33258 fluorescent staining.Apoptotic rate was quantified by flow cytometry (FCM) using Annexin-V/PⅠ dual staining.Western blotting was used to evaluate the changes of Fas,Caspase-8 and Caspase-3 proteins in U251 cells.Results PS Ⅰ and PS Ⅱ inhibted U251 cell proliferation with a timeand dose-dependent manner; as compared with that in the blank control group,the proliferation rate in the PSⅠ and PS Ⅱ treatment groups was significantly increased (P＜0.05); PS Ⅰ had stronger inhibited effect than PS Ⅱ.Typical morphological changes of apoptosis were observed in U251 cells of the PSⅠ and PS Ⅱ treatment groups with Hoechst 33258 staining.The early apoptotic rates of the PSⅠ and PS Ⅱ treatment groups were all significantly higher than that of the blank control groups (F=81.434,P=0.000).As compared with those in the blank control group,the expressions ofFas,Caspase-8 and Caspase-3 were significantly increased (P＜0.05).Conclusion PSⅠ and PS Ⅱ inhibite the proliferation ofglioma cells in vitro; they increase the expressions ofFas,Caspase-8 and Caspase-3,and accelerate the cell apoptosis,which might be the mechanism of inhibition.
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Objective:To understand the molecular pat hophysiology of hepatocellular carcinoma and pancreatic cancer.Methods: We studied novel gene expression by cDNA microarray method. The PCR pro ducts of 4 096 genes and 12 800 gene were spotted onto a kind of chemical-mater ial-coated-glass slide in array. Both the mRNAs from 5 cases of hepatocellular carcinoma and 3 cases of pancreatic cancer were reversely transcribed to cDNAs with the incorporation of fluorescent-labeled dUTP to prepare the hybridization probes. After hybridization, BioDoor4096 and BioDoor12800 cDNA microarray were scanned for the fluorescent intensity. Tumor invasion-related gene expression w as screened through the analysis of difference in gene expression profile.Results:Among 4 096 and 12 800 target genes, there were 15 genes who se expression level differed from normal and carcinoma tissues. Therefore, they might be associated with metastasis.Conclusion:Further analysis of these differentially expressed metastasis-associated genes will be helpful for understanding the molecular mechanism of malignant carcinoma.
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Objective:To understand the molecular pat hophysiology of hepatocellular carcinoma and pancreatic cancer.Methods: We studied novel gene expression by cDNA microarray method. The PCR pro ducts of 4 096 genes and 12 800 gene were spotted onto a kind of chemical-mater ial-coated-glass slide in array. Both the mRNAs from 5 cases of hepatocellular carcinoma and 3 cases of pancreatic cancer were reversely transcribed to cDNAs with the incorporation of fluorescent-labeled dUTP to prepare the hybridization probes. After hybridization, BioDoor4096 and BioDoor12800 cDNA microarray were scanned for the fluorescent intensity. Tumor invasion-related gene expression w as screened through the analysis of difference in gene expression profile.Results:Among 4 096 and 12 800 target genes, there were 15 genes who se expression level differed from normal and carcinoma tissues. Therefore, they might be associated with metastasis.Conclusion:Further analysis of these differentially expressed metastasis-associated genes will be helpful for understanding the molecular mechanism of malignant carcinoma.