GC-MS Analysis of Volatile Oil from Chaihu Guizhi Decoction and Study on Its Inhibitory Effects on in vitro Proliferation of Human Lung Adenocarcinoma A 549 Cells / 中国药房

OBJECTIVE：To study the compositi on of the volatile oil from Compound chaihu guizhi decoction ，and to evaluate its in vitro anti-proliferative activity on human lung adenocarcinoma A 549 cells. METHODS ：The volatile oil from Chaihu guizhi decoction was extracted according to the steam distillation method of general rules 2004 in the 2015 edition of Chinese Pharmacopoeia（part Ⅳ）. The volatile oil components were analyzed by GC-MS combined with Kováts index ，and the relative content of each component was calculated by peak area normalization method. Using different concentrations of cisplatin （4，8， 16，32，64 mg/L）as positive control ，MTT assay was used to detect the inhibitory effects of different concentrations of volatile oil from Chaihu guizhi decoction （25，50，100，200，400 mg/L）on in vitro proliferation of A 549 cell after 48 h of treatment. Negative control group （with cells but without drugs ）was set up. RESULTS ：A total of 71 chemical components were isolated from the volatile oil ，among which there were 59 compounds identified ，sum of peak areas accounting for 84.99％ of the total peak area. The compounds with relatively high content included ar-curcumene （17.65％），β-bisabolene（9.57％），β-ocimene（7.05％）， α-curcumene（5.35％），2，5-dimethylbenzaldehyde（4.24％），linalyl isobutyrate （2.70％），α-cedrene（2.48％），δ-cadinene （2.07％）. Compared with negative control group ，the proliferation rate of cells were decreased significantly in 4-64 mg/L cisplatin groups and 25-400 mg/L volatile oil from Chaihu guizhi decoction groups （P＜0.05）. IC 50 of cisplatin and volatile oil from Chaihu guizhi decoction to in vitro proliferation of A 549 cells were 10.150 and 73.526 mg/L. CONCLUSIONS ：The volatile oil from Chaihu guizhi decoction mainly includes ar-curcumene ，β-bisabolene，β-ocimene，α-curcumene，which shows certain inhibitory effect on in vitro proliferation of A 549 cells.

Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells

Spider venom is a potential source of pharmacologically important compounds. Previous studies on spider venoms reported the presence of bioactive molecules that possess cell-modulating activities. Despite these claims, sparse scientific evidence is available on the cytotoxic mechanisms in relation to the components of the spider venom. In this study, we aimed to determine the cytotoxic fractions of the spider venom extracted from Phlogiellus bundokalbo and to ascertain the possible mechanism of toxicity towards human lung adenocarcinoma (A549) cells. Methods: Spider venom was extracted by electrostimulation. Components of the extracted venom were separated by reversed-phase high performance liquid chromatography (RP-HPLC) using a linear gradient of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 95% acetonitrile (ACN). Cytotoxic activity was evaluated by the MTT assay. Apoptotic or necrotic cell death was assessed by microscopic evaluation in the presence of Hoechst 33342 and Annexin V, Alexa FluorTM 488 conjugate fluorescent stains, and caspase activation assay. Phospholipase A2 (PLA2) activity of the cytotoxic fractions were also measured. Results: We observed and isolated six fractions from the venom of P. bundokalbo collected from Aurora, Zamboanga del Sur. Four of these fractions displayed cytotoxic activities. Fractions AT5-1, AT5-3, and AT5-4 were found to be apoptotic while AT5-6, the least polar among the cytotoxic components, was observed to induce necrosis. PLA2 activity also showed cytotoxicity in all fractions but presented no relationship between specific activity of PLA2 and cytotoxicity. Conclusion: The venom of P. bundokalbo spider, an endemic tarantula species in the Philippines, contains components that were able to induce either apoptosis or necrosis in A549 cells.(AU)

Effect of cereblon on inhibition of human lung cancer A549 cell migration by thalidomide / 国际药学研究杂志

Objective:To investigate the influence of cereblon(CRBN)on the inhibitory effect of thalidomide on the migration of human lung adenocarcinoma A549 cells and related mechanism. Methods: The effect of thalidomide on the A549 cell proliferation was assayed by the MTS method. CRBN in A549 cells was knocked down(A549CRBN cells) with RNA interference and lentivirus infection. Transwell assay was performed to detect the effect of thalidomide on the cell migration of A549 and A549CRBN cells. RT-qPCR assay was performed to detect the gene expression of N-cadherin and vimentin. Results: The thalidomide at the 1, 10, 50 and 100 μmol/L concentration did not affect the proliferation of A549 cells, while the 100 μmol/L thalidomide could significantly inhibit the migration of A549 cells. After the CRBN gene knockdown the migration ability of A549 cells was significantly decreased(P<0.01), and the gene expression of N-cadherin and vimentin was also significantly reduced in the A549CRBN cells. The inhibitory effect of thalidomide on the cell migration in the A549 cells disappeared after CRBN gene knockdown. Thalidomide could inhibit the gene expression of N-cadherin and vimentin in a CRBN-dependent manner. Conclusion: Thalidomide could inhibit the migration of A549 cells and the expression of the migration-related genes via CRBN.

Expression of Acetaldehyde Dehydrogenase in Gefitinib-resistant Human Lung Adenocarcinoma HCC-827/GR Cells / 中国肺癌杂志

BACKGROUND@#Tumor recurrence and drug resistance are the main causes of death in tumor patients. The family of acetaldehyde dehydrogenase (ALDH) is closely related to the proliferation, migration, invasion and resistance of tumor cells, and different ALDH subtypes are expressed in different tumor cells. The aim of this study is to elucidate the ALDH subtype in human lung adenocarcinoma HCC-827/GR cells, which resistant to the gefitinib.@*METHODS@#The human lung adenocarcinoma HCC-827 cells were used to generate the gefitinib-resistant HCC-827/GR cells; the expression of ALDH subtype in either HCC-827 or HCC-827/GR was detected by flow cytometry; The proliferative capacity and sensitivity to gefitinib of hcc-827/GR cells were analyzed by MTT assay before and after treatment with 100 μmol/L diethyllaminaldehyde (DEAB); Real-time quantitative PCR was used to detect the expression of ALDH subtypes at mRNA levels in hcc-827 cells and hcc-827/GR cells.@*RESULTS@#Compared with HCC-827 cells, the positive rate of ALDH in HCC-827/GR cells increased. The proliferation ability of HCC-827/GR cells decreased after treatment with 100 μmol/L DEAB. Compared with HCC-827 cells, the expression of ALDH1A1 and ALDH1L1 mRNA was increased in hcc-827/GR cells, but the ALDH3B2 expression was decreased.@*CONCLUSIONS@#ALDH might be used as a molecular biomarker to test the gefitinib-resistant to lung adenocarcinoma cancer cells, and the ALDH1A1 may play a role in gefitinib resistance in lung cancer.

Protective effects of total flavonoids extraction from ficus lacor leaves on A549 cells / 重庆医学

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.

Effect of astragaloside IV on tumor cellular uptake and antitumor efficacy by ginsenoside compound K / 中草药

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.

Effects of Recombinant Adenovirus Carrying Apoptin Gene on the Apoptosis of Human Lung Squamous Carcinoma SK-MES-1 Cell Lines and Human Lung Adenocarcinoma NCI-H1299 Cell Lines / 中国药房

OBJECTIVE:To explore the effects of recombinant adenovirus vector Adxsi-GFP-VP3 carrying apoptin gene VP3 on the apoptosis of human lung squamous carcinoma SK-MES-1 cell lines and human lung adenocarcinoma NCI-H1299 cell lines. METHODS:The exponential phase SK-MES-1 and NCI-H1299 cell lines were respectively divided into a recombinant adenovirus (Adxsi-GFP-VP3) group,a empty virus (Adxsi-GFP) group and a cell control (phosphate buffer) group,which were marked as group A,B and C respectively. Reverse transcription polymerase chain reaction and Western blot method were used to detect the ex-pressions of VP3 mRNA and Apoptin in the cells of groups A and B 48 and 72 h after transfection. The change in the ultrastructure of the cells in group A was observed under transmission electron microscope 72 h thereafter. MTT method was adopted to detect the cell proliferation activities of three groups 24,48,72 and 96 h thereafter and flow cytometry to determine the apoptosis rates and cell cycle changes 24,48 and 72 h thereafter. RESULTS:Compared to group B,group A demonstrated the expression of VP3 mRNA in SK-MES-1 and NCI-H1299 cell lines 48 h after transfection,and Apoptin expression and ultrastructure change for apopto-sis of SK-MES-1 and NCI-H1299 cell lines 72 h thereafter. Compared to groups B and C,group A showed lower proliferation activ-ities and higher apoptosis rates of SK-MES-1 and NCI-H1299 cell lines,which had a positive correlation with transfection time;and in the group A,there was a decrease in the proportion of the SK-MES-1 and NCI-H1299 cell lines in S phase and an increase in the proportion of those in G2/M phase,72 h after transfection. There was statistically difference (P<0.05). CONCLUSIONS:Adxsi-GFP-VP3 can effectively induce the apoptosis of SK-MES-1 and NCI-H1299 cell lines.

Effect of ampelopsin sodium combined with carboplatin on the proliferation and apoptosis of human lung adenocarcinoma cell GLC-82 / 中国药理学通报

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.

Effect of rapamycin on proliferation, invasion, adhesion, apoptosis and autophagy of human lung adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum / 中国病理生理杂志

[ ABSTRACT] AIM:To study the effect of rapamycin ( Rap) on the proliferation, invasion, adhesion, apoptosis and autophagy of human adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum ( DDP) .METHODS: Human adenocarcinoma A549 and resistant A549/DDP cell lines were cultured.The inhibitory effects of Rap alone or combined with DDP on A549 and resistant A549/DDP cells were detected by MTT assay.The in vitro invasion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by Transwell methods. The in vitro adhesion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by adhesion experiments.The apoptosis of A549 and resistant A549/DDP cells induced by Rap alone or combined with DDP was ana-lyzed by flow cytometry.The cell autophagy marker proteins beclin-1 and LC3 in A549 and resistant A549/DDP cells trea-ted with Rap alone or combined with DDP were detected by Western blotting.RESULTS:Compared with Rap or DDP a-lone group, the combination of Rap and DDP significantly inhibited the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells in vitro, and promoted the cell apoptosis and autophagy marker proteins beclin-1 and LC3 expres-sion ( all P<0.05) .CONCLUSION:Rap enhances the effect of DDP through promoting the cell autophagy, thereby in-hibiting the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells and inducing the cell apoptosis with a synergistic effect.

The Inhibitory Effects of Rh-endostatin(YH-16)in Combination with Radiotherapy on Lung Adenocarcinoma A549 in Mice and the Underlying Mechanisms / 华中科技大学学报（医学）（英德文版）

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.

Effect on telomere of antisense tankyrase and telomerase oligonucleotide in human lung adenocarcinoma A549 cell line / 基础医学与临床

Objective To determine the effect of transcription and translation in telomeric related proteins,and synergism of progressive telomere shortening and cell cycle alteration in human lung adenocarcinoma A549 cell line,which is induced by antisense tankyrase oligonucleotide(asTANKS) combinated with antisense human telomerase reverse transcriptase(ashTERT) oligonucleotide.Methods A549 cells were randomly assigned as 3 test groups: ashTERT,ashTERT + asTANKS and asTANKS,three control groups(shTERT,sTANKS and blank).With individual intervention for different hours,the effect of transcription in hTERT mRNA was evaluated by RT-PCR,and telomerase activity was tested by ELISA-PCR,tankyrase activity was tested by Western blot as well.Moreover,telomere average length was analyzed by Q-FISH,and duration of proliferation was observed by population double test.Results Transcription in hTERT mRNA and telomerase activity for 48 hrs was inhibited obviously by ashTERT,but not by asTANKS.Progressive telomere shortening in A549 cells for 48 hrs was induced by either asTANKS or ashTERT(vs control,P

Transfection of T-STAR antisense gene can depress the telomerase activity in lung adenocarcinoma cell line A549 / 第三军医大学学报

Objective To investigate the changes of telomerase activity after knocking down endogenous expression of T-STAR (testes-signal transduction and activator of RNA) gene in human lung adenocarcinoma cell line A549 by antisense strategy. Methods The mRNA and protein expression of T-STAR gene were determined by RT-PCR and Western blotting, and the telomerase activity was measured by PCR-ELISA, after transfection of T-STAR antisense gene into A549 cells with lipofectamine. Sense pcDNA-STAR and blank pcDNA3.1 transfection served as control. Results The expression of T-STAR gene was significantly inhibited at mRNA and protein level, and the telomerase activity was significantly decreased. Conclusion The down-regulation of telomerase activity may result from inhibition of T-STAR gene expression in lung adenocarcinoma A549 cells.

Effects of Qingjin Desheng Tablets on Telomerase and Protein Expressions of Apoptosis Relevant Genes in Human Lung Adenocarcinoma Cells / 中药新药与临床药理

Objective To explore the molecular mechanism of Qingjin Desheng Tablets on inhibiting the growth of lung cancer cells and to supply experimental evidences for Chinese herbal medicine preventing and curing lung cancer in clinical application .Methods Immunohistochemistry staining was used to detect the activity of telomerase and the expression of bcl-2 and bax in human lung adenocarcinoma cells after treated with serum containing Qingjin Desheng Tablets.Results The activity of telomerase was inhibited significantly and the positive rate of bcl-2 protein reduced,and the positive rate of bax2 increased in human lung adenocarcinoma cells after 72 hour-treatment with the serum containing Qingjin Desheng Tablets.Conlusion Qingjin Desheng Tablets exert the anti-tumor action through inhibiting the activity of telomerace and modulating the expression of bcl-2 and bax protein.

Sensitization effect of insulin of chemotherapy on human adenocarcinoma cells A549 / 解放军医学杂志

Objective To explore the sensitization effect of insulin to chemotherapy as a metabolic promoter on human lung adenocarcinoma cells A549.Methods The sensitivity of human adenocarcinoma cells A549 to DDP was observed by MTT methods,and IC30 values of the cells were calculated.The optimal concentration of insulin to promoting A549 cells' growth was determined by MTT assay.The A549 cells were pretreated with insulin of various concentrations for different time period,and then the cell sensitivity to DDP was evaluated with MTT assay.Flow cytometry was used to study the cell cycle distribution of the A549 cells treated with insulin.Results The IC30 value of DDP was about 36.87?g/ml.It was found that the cell growth was significantly promoted with pretreatment of insulin at the concentration of 4~16mU/ml for 8~16 hours.Insulin enhanced the inhibitory effect of DDP(36.87?g/ml)on cell proliferation when human lung adenocarcinoma cells A549 were treated with insulin(2.0~16.0mU/ml;8~16h)as indicated by MTT assay(P