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Objective:To investigate the change of apoptosis-associated genes in human lung adenocarcinoma cell lines A549/DDP cells,which were induced by the recombinant human interleukin-24(rhIL-24) combined with Cisplatin (DDP). Methods: Six genes expression level by GeXP genetic analysis system at the same time,after rhIL-24,DDP and rhIL-24+DDP were used to intervene in A549/DDP cells. Results:rhIL-24 could induce Bax gene,Caspase3 gene and Rb gene transcription up regulation;Bcl-2 gene and survivin gene transcription down regulation. But no regular change in the genes expression level of P53. Bax,Survivin and Rb were more obviously changed after rhIL-24 combined with DDP. Conclusion: RhIL-24 can induce apoptosis of A549/DDP cells through upregulated the genes expression level of Bax,Caspase3,Rb and down regulated of Bcl-2,Survivin.
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Objective To explore the effects of all-trans-retinoic acid (ATRA) on the proliferation of human lung adenocarcinoma cell line A549 and the expression of APLNR (apelin receptor) gene.Methods The inhibition of proliferation of human lung adenocarcinoma cell lines A549 cultured in vitro with or without ATRA was measured by MTT (methyl thiazolyl tetrazolium,MTT) method.The morphological changes in the cells were observed by light microscopy.The cell cycle and apoptosis were analyzed by flow cytometry.The levels of APLNR,cyclin D1 and p16 proteins were detected by western blot.Results After treatment of ATRA,the proliferation of A549 cells was obviously inhibited in dose-and time-independent manner (P ＜ 0.01).The cell morphology was significantly changed.The cycle of A549 cells was blocked at G0/G1 phase and the apoptosis rate was increased.With the increasing concentration of ATRA,the expressions of cyclin D1 and APLNR were down-regulated but the expression of p16 was up-regulated (P ＜ 0.01).Conclusion ATRA could inhibit the proliferation of A549 cells by retardant cell cycle of A549 cells at G0/G1 phase and inducing the apoptosis,and down-regulate the expression of APLNR gene.
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Objective To investigate the extraction method of total flavonoids from the leaves of ficus lacor and the protec tive effects of extraction on the cellular damage to provide a basis for the research on the phamaceutical value of ficus lacor leaves.Methods The ethanol extraction method was adopted to extract the total flavonoids in the leaves of ficus lacor and the extraction efficiency was calculated with rutin as the standard.The rotenone induced human lung adenocarcinoma cellular damage served as the model,then the influencesof the extraction on the cellular viability,cellular morphology,production of reactive oxygen species (ROS) and apoptosis were researched.Results The extraction efficiency of total flavonoids in the leaves of ficus lacor by 60％ ethanol was 5.02％;the extraction at the concentration of 32 mg/L could significantly inhibit the decrease of cell viability,cellular shape change,ROS production and apoptosis of A549 cells induced by 100μg/L rotenone.Conclusion The ethanol extraction method can be used to extract the total flavonoids in the leaves of ficus lacor and the extraction has the protective effects on the A549 cellular dam age induced by rotenone,the leaves of ficus lacor have the potential for further researching its pharmaceutical value.
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Objective: To investigate the effect of astragaloside IV on tumor cellular uptake and antitumor efficacy by ginsenoside compound K (GCK). Methods: The human lung adenocarcinoma cell line A549 was prepared as an antitumor model, the cytotoxicity of the mixtures of GCK and astragaloside IV to A549 cells was determined by MTT assay, the cellular uptake of GCK was detected by fluorescence microscopy, and the intracellular GCK was determined by HPLC. The enhancement of astragaloside IV in vivo efficacy by GCK was evaluated by nude mice bearing tumor model. Results: The effect of GCK on the inhibition of A549 cell proliferation was enhanced in the presence of astragaloside IV and astragaloside IV could increase the uptake rate of GCK in A549 cells, with the proportion of astragaloside IV increasing, the cytotoxicity was significantly stronger than GCK and the uptake rate was improved. In vivo antitumor assay of mice bearing tumor indicated that astragaloside IV could enhance the antitumor efficacy of GCK. Conclusion: Preliminary results indicate that the mixture of GCK and astragaloside IV have the effect of enhancing antitumor efficacy.
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Purpose To investigate the application value in diagnosis of pleural effusion by cell block combined with immunohisto-chemistry. Methods 60 cases of pleural effusion were collected, and paraffin-embedded cell block was prepared and immunohisto-chemistry was used to detect the expression of CK7, TTF-1, E-cadherin, CEA and Calretinin. Results By use of cell block combined with immunohistochemistry, malignant detected rate was higher than that of the conventional centrifugal smear. There was statistical significance in the expression of CK7, TTF-1, E-cadherin, CEA and Calretinin between pleural effusion lung adenocarcinoma and reac-tive hyperplasia mesothelial cells (P<0. 05). CK7, TTF-1, E-cadherin and CEA were highly expressed in pleural effusion of lung ad-enocarcinoma cell. Calretinin was highly expressed in hyperplastic mesothelial cells. Conclusion Cell block and immunohistochemi-cal technique combination in the differential diagnosis of difficult pleural effusion has important clinical significance. It is worthy of popularization and further clinical application.
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Aim To investigate the cytotoxic effect and mechanism of ampelopsin sodium ( AMP-Na ) on hu-man lung adenocarcinoma cell line GLC-82 alone or combined with carboplatin ( CBP ) . Methods The cytotoxic effect of human lung adenocarcinoma cell line GLC-82 was investigated by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide ( MTT ) colori-metric assay. Ultrastructure change of apoptotic GLC-82 cells was observed with transmission electron micro-scope. The changes of the cell apoptosis and the ex-pression of caspase-3 were analyzed with flow cytome-ter. Results Combined with AMP-Na, the IC50 of CBP decreased from (17. 10 ± 4. 78) mg·L-1 to tron microscope and flow cytometric analysis, the apop-tosis and necrosis ratios also increased in the combina-tion group. The necrosis ratios increased from (2. 56 ± 0. 41 )% to ( 71. 83 ± 5. 43 )% ( P<0. 01 ) . The ex-pression of caspase-3 was increased significantly after treated with AMP-Na or combined with CBP. Conclu-sions There is a synergistic cytotoxic effect on GLC-82 cells treated with AMP-Na combined with CBP. Ap-optotic cells and necrotic cells are found in GLC-82 cells treated with AMP-Na alone or combined with CBP. One of the mechanisms to induce apoptosis is probably that activation of caspase-3 mediates signal transduction pathway in cells.
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Objective To purify recombinant methioninase and investigate the synergetic inhibitory effect of methioninase and cisplatin on the proliferation of human lung adenocarcinoma cell line GLC. Methods Recombinant methioninase was purified with GST-column from supernatant after ultrasonic disruption of cultured Escherichia coli in the prokaryotic expression system pGEX-4T-1-Met/Dh5a. MTT assay was used to determine the inhibition rate of methioninase in combined with cisplatin on cell proliferation,and their synergistic effect was evaluated by using the q value judge method. Results The concentration of recombinant methionine cleaving enzyme was 0.22 mg/mL, the purity was 95%, and the activity was 0.568 IU/mg. After 72 h culturing, the inhibition rate of cisplatin methionine was 24.80%and 27.49%respectively,while the inhibition rate of the combined drugs was 66.80% ( =1.47>1.15) which showed a significant synergistic effect. Conclusion Both methioninase and cisplatin have the inhibition effect,and the combined drugs display a significant synergistic effect on the proliferation of GLC.
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Objective To study the radioseusitization effect of gold nanoparticles modified by sodium glycididazole.Methods The sodium glycididazole was connected to gold nanoparticle,in dimension of about 18 nm,that had been modified with polyethylene glycol.The nanoparticle-swallowing efficiency of lung adenocarcinoma A549 cells was observed by a scanning electron microscope.Cells were divided into four groups:sodium glycididazole group,gold nanoparticles group,sodium glycididazole-gold nanoparticles group,and no drug control group.The radiosensitivity was detected by MTT and colony formation assays.Results Sodium glycididazole-gold nanoparticles could enter the cell cytoplasm and nucleus.The concentration of 0.003 mg/ml gold nanoparticles and sodium glycididazole-gold nanoparticles had no obvious cytotoxic effect.After irradiation of 2,4,6,8 Gy,the cell survival of the sodium glycididazole-gold nanoparticle group was significantly lower than that of the other three groups (F =4.8,14.5,5.7,7.6,P ＜0.05) and the D0 and Dq values of the sodium glycididazole-gold nanoparticle group were significantly lower than those of other three groups.Conclusion Sodium glycididazole-gold nanoparticles can enhance the radiosensitivity of lung adenocarcinoma cells.
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In order to investigate the inhibitory effects of Endostar(rh-endostatin,YH-16)in combination with radiotherapy on lung adenocarcinoma A549 in mice and the interaction mechanisms of combined therapy,the transplantation tumor models of A549 lung adcnocarcinoma were established.When the largest diameter of tumor reached 1.0 cm,all nude mice were randomly divided into 4 groups: Endostar group,radiotherapy group,radiotherapy plus Endostar(combined treatment)group,and control group(n=6 in each group).The largest diameter and the vertical diameter of tumor were measured at different time points.At the 16th day,mice were executed,and the tumors were applied to analysis of rate of tumor cell apoptosis,and the expression levels of basic fibroblast growth factor(bFGF)mRNA were detected by reverse transcription-polymerase chain reaction(RT-PCR)and those of vascular endothelial growth factor(VEGF)by immunohistochemistry.The results demonstrated that the rate of tumor inhibition in combined treatment group was higher than that in other groups.And the rate of tumor cell apoptosis in combined treatment group was also higher than that in other groups.Meanwhile,the levels of bFGF mRNA and VEGF expression in combined treatment group were lower than those in other groups.It was concluded that Endostar obviously enhanced the curative effectiveness of radiotherapy on lung adenocarcinoma A549 in mice.The underlying mechanisms may involve the down-regulation of bFGF mRNA and VEGF expression to inhibit angiogenesis by Endostar and the cooperative effect of Endostar and radiotherapy to synergistically promote tumor cell apoptosis.And Endostar inhibits angiogenesis by down-regulating the expression of bFGF mRNA and VEGF.
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Objective:To investigate the radiosensitization and the cell-cycle of mild hyperthermia(≤42℃)on human pulmonary adenocarcinoma cell line SPC-A-1 in vitro. Methods: The human pulmonary adenocarcinoma cell line SPC-A were treated with radiation and the combination of radiation with mild hyperthermia. Radiosensitivity was determined by clonogenic assay and quantified by calculating the thermal enhancement ratio (TER). Flow cytometry was used to observe the cell-cycle. Results: Do, Dq calculated from the dose-response curve for radiation combined with 41.5℃ were 1.390 Gy, 1.426 Gy, whereas 1.693 Gy, 2.453 Gy for radiation alone, respectively. TER was 1.218. The proportion of cells in S phase was found to be 14.81% in the radiation group. The values, after 48 hours and 72 hours, with 6Gy radiation combined immediate 41.5℃ one hour mild hyperthermia, were 5.89% and 9.08%, respectively, versus 18.8% and 31.91% with 6 Gy radiation alone. Conclusion:Radiosensitization of mild hyperthermia in SPC-A-1 cells associated with the hyper-radiosensitization of the cells in S phase.
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Objective To study the effect of the recombinant matrilysin (rMMP-7) and its antisense oligonucleotide transfected by liposome (LIPO-MAON) on the proliferation of lung adenocarcinoma cell line A549 and human umbilical vein endothelial cells (HUVEC). Methods Antisense oligonucleotide of matrilysin was constructed by liposome transfection. The proliferation of HUVEC and A549 was detected by MTT assay in case of rMMP-7 or LIPO-MAON existence. The expression of MMP-7 mRNA in both HUVEC and A549 was examined by semi-quantitative RT-PCR assay. Results The proliferation of both HUVEC and A549 cell line was accelerated by high level of rMMP-7 (0.75, 1.0 ?g/ml), and the inhibited proliferation was only found in A549 cell line treated with high concentration of LIPO-MAON (7.5, 10 nmol/ml), but not in HUVEC. By RT-PCR assay, no expression of MMP-7 mRNA was found in HUVEC; however, enhanced or decreased expression of mRNA were found when A549 cell line was treated respectively with rMMP-7 or LIPO-MAON (P
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Objective:To investigate the inhibitory effects of cytokine induced-killer(CIK) cells combined with docetaxel(DTX) on the expansion of lung adenocarcinoma cell line SPC-A1/DTX in vitro and in vivo.Methods:Peripheral blood mononuclear cells(PBMC) from healthy donors were incubated in vitro to induce CIK cells in the presence of interferon-gamma(IFN-?),IL-2 and anti-CD3 monoclonal antibody.MTT assay was employed to evaluate the cytotoxic activity of DTX,CIK cells and their combination against SPC-A1/DTX cells in vitro.SPC-A1/DTX lung adenocarcinoma cells were injected subcutaneously into nude mice.On the 14 th day,normal saline(control group),docetaxel(DTX 1 mg/kg in 0.2 ml,DTX group),CIK cells(1?107,CIK group),and CIK cells combined with docetaxel(CIK+DTX group) were administered intraperitoneally respectively.All the nude mice were sacrificed at day 15 after treatment and the tumor were weighted out.Results:MTT assay showed that the IC50 to docetaxel in SPC-A1/DTX cells was 110.5 ?g/ml and 8.5 ?g/ml in wild SPC-A1 cells.CIK cells possessed a higher anti-tumor cytotoxic activity in SPC-A1/DTX cells in vitro than in wild SPC-A1 cells(P
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Objective To observe the effects of arsenic trioxide(As2O3)on the expressions of TNF-?,Fas and bcl-2 in lung adenocarcinoma cells(LAC)and to explore the mechanism of arsenic trioxide inducing apoptosis.Methods The expressions of TNF-?,Fas and bcl-2 in lung adenocarcinoma cells pretreated by arsenious acid were determined by the double antibody sandwich ABC-ELISA method.Results Compared with the control group,As2O3 showed no effects on the contents of bcl-2 in lung adenocarcinoma cells after 72 hours treatment,but increased the contents of TNF-? and Fas significantly,and the effects in different concentration groups had significant differences.The protein expressions of TNF-? and Fas showed a tendency of concentration-dependent increasing.Conclusions The results suggest that As2O3 induces the apoptosis of LAC cells possibly by up-regulating the expression of TNF-? and Fas.
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AIM To study the effects of ASA on proliferation and apoptosis of human lung Adenocarcinoma cell lines SPCA-1 in vivo. METHODS Cytotoxicity assay was tested by MTT method.Cell cycle was analyzed by flow cytometry(FCM).The morphology of the treated cells was observed by wright exclusion,Hoechest/PI exclusion, electron microscope. Apoptosis landder was evaluated by agarose gel electrophoresis of DNA. RESULTS ASA inhibited SPCA-1 cell proliferation in a time-and dose-dependent fashion(1.0～12.5 mmol?L -1,24 h～72 h). ASA increased the number of cells in G 0/G 1 and G 2/M phases,degrased the population of S phases at on 24 h and incresed apoptosis cells number. CONCLUSION ASA may inhibit the proliferation of SPCA-1 cell lines through effects on cell cycle and apoptosis.
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Objective To investigate the possibility of inhibiting the malignant proliferation of tumor cells in vitro through down-regulating the expression of MMP-7 by antisense gene technology. Methods A Phosphorothioate antisense oligodeoxynucleotide (ASODN) and scrambled oligodeoxynucleotide (SCODN) against MMP-7 were respectively transfected into A549 cells mediated by liposome. The expression of MMP-7 and cell proliferation in the cells were respectively examined by immunohistochemical assay and MTT. Flow cytometry was used to detect the cell cycles. Results After A549 cells were transfected with MMP-7 ASODN, the cell number was decreased. MTT method indicated that the proliferation of A549 cells was inhibited by MMP-7 ASODN at different concentrations, with the inhibition reaching the peak at 48 h. ASODN transfection downregulated the expression levels of MMP-7 protein, and inhibited the transition period of G_ 0 /G_ 1 phase to S phase. Conclusion The artificial MMP-7 ASODN can specifically inhibit the expression of MMP-7 protein and regulate cell cycle and cell proliferation in A549 cells.