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Aim To observe the effect of methylation inhibitors of 5-aza-2'-deoxycytidine (5-Aza-CdR) on SPC-Al - lung cancer cell proliferation, cell scratches and apoptosis, to explore the influence of secretion curl associated protein 1 (SFRPl) and 06-methylguanine-DNA-methyltransferase (MGMT) gene promoter region in which DNA methylation mRNA and protein expression and meaning. Methods The effects of 5-Aza-CdR at different concentrations on the proliferation of human lung cancer SPC-A 1 cells were determined by CCK-8 assay. The effect of 5-Aza-CdR on the migration ability of SPC-A 1 cells was determined by scratch assay. The apoptosis of lung cancer SPC-A 1 cells was detected by Hoechst 33258 staining after treatment with 5-Aza-CdR for 24 h. mRNA and protein expressions of SFRPl and MGMT in SPC-Al cells were detected by RT-PCR and Western blot. Results 5-Aza-CdR could reduce the proliferation of SPC-A 1 cells by concentration gradient,and IC50 was 21.2 jjimol • L
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OBJECTIVE： To investigate the mechanism of calycosin （CA） inhibiting the proliferation and migration of lung adenocarcinoma cells by regulating miR-21/PTEN signaling pathway. METHODS： Using lung adenocarcinoma SPC-A1 cells as objects， cell proliferation was detected by MTT method after treated with different doses of CA （5， 15， 25， 50， 75， 100 μg/mL） for 12， 24， 48， 72 h. Cell survival rate， 30％ cell growth inhibition concentration （IC30） and half inhibition concentration （IC50） were calculated. Transwell migration test was used to detect the migration of cells after treated with low-dose， medium-dose and high-dose of CA （50， 75， 100 μg/mL） for 24 h. The number of stained cells was recorded and inhibition rate of cell migration were calculated. Western blotting assay and real-time PCR were used to detect the expression of miR-21 as well as the proteins and their mRNAs expression of PTEN， VEGF， MMP-9 after treated with low-dose， medium-dose and high-dose of CA （50， 75， 100 μg/mL） for 24 h. After transfected with miR-21 mimics and miR-21 inhibitor， the effects of CA （75 μg/mL） on the expression of miR-21 and the protein expression of PETN， VEGF and MMP-9 were detected. RESULTS： After treated with 50， 75， 100 μg/mL CA for 12， 24， 48 h， 25， 50， 75， 100 μg/mL CA for 72 h， cell survival rate was decreased significantly （P＜0.05 or P＜0.01）. IC30 of CA were 82.24， 50.45， 46.34， 31.81 μg/mL ； IC50 of CA were 108.06， 73.35， 70.08， 49.89 μg/mL during 12-72 h. Compared with normal control group， the number of stained cells in CA groups， protein expression of VEGF in CA low-dose group， expression of miR-21 as well as proteins and their mRNAs expression of VEGF， MMP-9 in CA medium-dose and high-dose groups were decreased significantly； the medium-dose and high-dose groups were significantly less or lower than low-dose group； the high-dose group was significantly less or lower than medium-dose group （P＜0.05 or P＜0.01）. Cell migration rate of CA groups as well as protein and its mRNA expression of PTEN in CA medium-dose and high-dose groups were increased significantly； the medium-dose and high-dose groups were significantly higher than the low-dose group； the high-dose group was significantly higher than the medium-dose groups （P＜0.05 or P＜0.01）. After transfected with miR-21 mimics， expression of miR-21 as well as protein expression of VEGF and MMP-9 were increased significantly in miR-21 mimic group， compared with normal control group； protein expression of PTEN was decreased significantly （P＜0.01）. After intervened by CA， expression of miR-21 as well as protein expression of VEGF and MMP-9 in cells were decreased significantly， compared with miR-21 mimic group； protein expression of PTEN was increased significantly （P＜0.05 or P＜0.01）. After transfected with miR-21 inhibitor， expression of miR-21 as well as protein expression of VEGF and MMP-9 were decreased significantly in miR-21 inhibitor group， compared with normal control group； protein expression of PTEN was increased significantly （P＜0.05 or P＜0.01）. After intervened by CA， the expression of miR-21 and above protein had no significant change in cells， compared with miR-21 inhibitor group （P＞0.05）. CONCLUSIONS： CA can inhibit the proliferation and migration of lung adenocarcinoma SPC-A1 cells in a dose-dependent manner， which may be associated with the regulation of miR-21/PTEN signaling pathway.
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OBJECTIVE:To study the antitumor activity of Anthopleura xanthogrammica crude extract on human lung cancer SPC-A1 cells in vitro. METHODS:A. xanthogrammica crude extract obtained by the methods of repeated freezing and thawing,ac-etone precipitation. After treated with crude extract 0(blank control),0.625,1.25 and 2.5 mg/ml for 24,48 and 72 h,the activity of SPC-A1 cells were measured by MTT assay. The growth inhibition rate and IC50 were also calculated. 24 h later,the morphologi-cal changes of SPC-A1 cells were observed by HE staining and AO/EB fluorescence staining. RESULTS:MTT assay showed that A. xanthogrammica crude extract has significant inhibitory effect on the proliferation of human lung cancer SPC-A1 cells;with the increasing of the concentration and the extension of the time,the inhibitory rate was increased. Its 24 h,48 h ,72 h IC50 were 1.81,1.32 and 1.18 mg/ml. HE staining and AO/EB staining appeared obvious morphological changes of apoptosis that cell mor-phology narrowed,vacuoles arose in the cytoplasm,karyopyknosis and part of nuclear disappearance occurred. CONCLUSIONS:A. xanthogrammica crude extract has an inhibitory effect on the proliferation of human lung cancer SPC-A1 cells.
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AIM@#In an effort to identify novel, small molecules which can affect the proliferation of lung cancer cells, F-01A, a polyether antibiotic isolated from the fermentation broth of Streptomyces was tested.@*METHOD@#F-01A was tested for its antitumor properties on the lung cancer cell line SPC-A-1, at six doses (0.1, 0.5, 1, 2.5, and 5 μmol·L(-1)), using various cellular assays. Cell viability was measured by the MTT assay, Hochest 33258 was used to study nuclear morphology; DNA ladder and the loss of mitochondrial membrane potential were also evaluated.@*RESULTS@#F-01A induces apoptosis against SPC-A-1 cells in a dose-dependent manner. The IC50 is 0.65 μmol·L(-1), and the inhibition at 5 μmol·L(-1) is 87.89%. Further, JC-1 staining indicates F-01A could induce the loss of mitochondrial membrane potential, and the DNA fragment is evident.@*CONCLUSION@#Mechanistic analysis showed that F-01A induced apoptosis of cancer cells probably in the mitochondrial pathway. The antitumor actions of F-01A involve activation of the apoptotic pathway against SPC-A-1 cells, and it may be valuable for further drug development.
Subject(s)
Humans , Anti-Bacterial Agents , Metabolism , Pharmacology , Apoptosis , Cell Line, Tumor , Cell Proliferation , Cell Survival , Growth Inhibitors , Pharmacology , Lung Neoplasms , Membrane Potential, Mitochondrial , Streptomyces , Chemistry , MetabolismABSTRACT
OBJECTIVE: To study the effect of various polarity fractions extracted from Sancaofang (SCF) and its combination on proliferation of human lung adenocarciaoma SPC-A-1 cells for bolting the most effective position of anti-tumor and suitable compatibility regimes.METHODS: Ethanol extraction and water extraction were adopted to prepare 95%,60% and 30% ethanol extract portion,water extract portion of SCF and compound decoction.The MTT assay was used to determine the effect of various polarity fractions of SCF,decoction and its combination on SPC-A-1 cells proliferation.RESULTS: IC50 of 60% ethanol extract was the smallest for SPC-A-1 cells.60% ethanol extract combined with 95% ethanol extract acts as a stimulus to anti-tumor activity significantly.CONCLUSION: The best suitable compatibility regimes were 95% ethanol extract combined with 60% ethanol extract.The liposolubility extract of SCF can be applied for anti-tumor.
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OBJECTIVE:To explore the effects of ethanol extracts of Solanum lyratum Thunb(ST) on induction of apoptosis and the expression of apoptosis associated genes fas and caspase-3 in human lung cancer SPC-A-1 cells.METHODS:Cultured human lung cancer SPC-A-1 cells were randomly divided into the control group,ethanol extracts of ST treated groups(2.5、5、10 mg?L-1)and the positive control group(cisplatin).After treatment with drug for 48 h,the proliferation inhibitory rate was evaluated by MTT assay,induction of cell apoptosis rate was determined by TUNEL method,the expression of fas and caspase-3 mRNA was detected by semi-quantitive RT-PCR.RESULTS:Compared with control group,the inhibitory rate was increased obviously(P
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Objective:To construct an eukaryotic radiation-inducible expressing vector of the human perforin N-terminal(hPFN-N),and to investigate the distribution and the killing effect of human perforin N-terminal truncated 118 amino acid polypeptide (rhPFN-N,22-139aa) on tumor cells.Methods:The gene hPFN-N was amplified by PCR from the plasmid pcDNA3.1(+)/hPFN and an enkaryotic radiation-inducible expression vector pcDNA3.1(+)/ Egr-hPFN-N was constructed after DNA recombination.After transfecting SPC-A1 cells with this recombination vector via liposome mediation,the expression of the hPFN-N protein was detected by RT-PCR and Immunocytochemical method and the killing effect of hPFN-N protein was assessed by standard MTT chromatometry.Results:DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic radiation-inducible expressing vector pcDNA3.1(+)/ Egr-hPFN-N had been constructed successfully.After the recombinant plasmid being transfected into SPC-A1 cells and being irradiated by X ray,RT-PCR verified the expression of hPFN-N mRNA.The result of Immunocytochemical assay was positive and in MTT assay the killing activity of rhPFN-N on target cells was 29.2%.Conclusion:After being irradiated the hPFN-N gene is expressed on the cell membrane and the killing activity of rhPFN-N on target cells is 29.2%.