Evaluation of arjunolic acid against Brucella melitenis and in vitro cytotoxic study of lung adenocarcinomic cell line (A549)

Brucellosis, a neglected tropical disease of zoonotic nature, is caused by the genus Brucella, specifically by Brucella abortus and B. melitensis in cattle and humans, respectively. Arjunolic acid (AA) is a triterpenoid, isolated from Terminalia arjuna (Roxb.) Wight & Arn., a medicinally important plant used to treat various diseases in the Indian system of medicine. Here, we tried to evaluate AA for its antibacterial activity on Brucella and the in vitro cytotoxicity assay on human lung adenocarcinomic alveolar basal epithelial cell line (A549). Also, we assessed the synergistic effect of arjunolic acid and Tarenna asiatica (L.) Kuntze ex K.Schum. on B. melitensis. AA displayed a considerable antibacterial activity [zone of inhibition (9 mm) with a minimum inhibitory concentration of 30 ?g/mL] against B. melitensis. The rate of cell death for the cancer cells were at 100 ?g/mL concentration of AA was 82% which indicates that AA shows significant membrane disruption to cancer cells. The estimated IC50 of AA against the A549 cell line was 139.90 ?g/mL. The highest synergistic activity was exhibited forming a zone of inhibition measuring 10mm when arjunolic acid and AqE of T. asiatica was added in the concentration of 1:1, respectively.

In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

circ-PRKDC affects the proliferation, apoptosis and radiosensitivity of lung cancer cells by regulating miR-505-3p / 中华放射肿瘤学杂志

Objective:To investigate the effect of circ-PRKDC on lung cancer cell proliferation, apoptosis and radiosensitivity and its molecular mechanism.Methods:Normal lung epithelial cells BEAS-2B and lung cancer cell lines NCI-H1299, NCI-H2170, NCI-H1975 were cultured. NCI-H1299 cells were divided into the si-NC, si-PRKDC, pcDNA-NC, pcDNA-PRKDC, miR-NC, miR-505-3p, anti-miR-NC, anti-miR-505-3p, si-PRKDC+ anti-miR-NC and si-PRKDC+ anti-miR-505-3p groups. RT-qPCR was used to detect the expression levels of circ-PRKDC and miR-505-3p. Western blot was employed to measure the protein expression. MTT was used to detect cell proliferation. Flow cytometry was utilized to detect cell apoptosis. Plate clone formation assay was conducted to detect the cell radiosensitivity. Dual luciferase reporter assay was performed to analyze the targeting relationship between circ-PRKDC and miR-505-3p.Results:Compared with normal lung epithelial cells BEAS-2B, the expression levels of circ-PRKDC in the lung cancer cell lines NCI-H1299, NCI-H2170 and NCI-H1975 were significantly up-regulated (3.65, 3.10, 2.67 vs. 1.00, all P<0.05), whereas those of miR-505-3p were significantly down-regulated (0.42, 0.50, 0.54 vs. 1.02, all P<0.05). After low expression of circ-PRKDC, the expression level of CyclinD1 was significantly down-regulated (0.42 vs. 0.81, P<0.05), whereas those of Cleaved-caspase-3(0.71 vs. 0.33, P<0.05) and γ-H 2AX (0.89 vs. 0.46, P<0.05) were significantly up-regulated, the cell A value was significantly decreased (0.413 vs. 0.839, P<0.05), cell apoptosis rate was significantly increased (20.35 vs. 6.21, P<0.05), cell survival fraction was significantly decreased ( P<0.05), and β-catenin expression was significantly down-regulated (0.35 vs. 0.73, P<0.05). After high expression of miR-505-3p, the expression level of CyclinD1 was significantly down-regulated (0.34 vs. 0.83, P<0.05), those of Cleaved-caspase-3(0.65 vs. 0.32, P<0.05) and γ-H 2AX (0.96 vs. 0.45, P<0.05) were significantly up-regulated, the cell A value was significantly decreased (0.386 vs. 0.851, P<0.05), the apoptosis rate was significantly increased (16.38 vs. 6.20, P<0.05), and the cell survival fraction was significantly decreased ( P<0.05). Compared with miR-NC, the luciferase activity of miR-505-3p group transfected with circ-PRKDC wild-type reporter plasmid was significantly decreased (0.44 vs. 1.00, P<0.05). Down-regulation of miR-505-3p could reverse the effect of low expression of circ-PRKDC on the proliferation, apoptosis, radiosensitivity and β-catenin expression of NCI-H1299 cells. Conclusion:Low expression of circ-PRKDC may inhibit lung cancer cell proliferation, promote cell apoptosis and enhance cell radiosensitivity by up-regulating miR-505-3p, which is probably associated with the Wnt/β-catenin signaling pathway.

Effect and possible mechanism of Salvianolic acid B on radiosensitivity of non-small cell lung cancer / 中华放射肿瘤学杂志

Objective:To evaluate the effect of Salvianolic acid B on the radiosensitivity of human non-small cell lung cancer cells and investigate its possible mechanism.Methods:Non-small cell lung cancer cells A549 and H1299 were cultured in vitro. The toxicity of Salvianolic acid B on non-small cell lung cancer cells was detected by MTT assay. The effect of Salvianolic acid B on the radiosensitivity was assessed by clone formation assay. Transwell chamber assay was used to evaluate the effect of Salvianolic acid B on the migration of tumor cells. Western blot was employed to detect the expression levels of OTUD7B, MMP-2, MMP-9, E-cadherin, Akt and p-Akt regulated by Salvianolic acid B. Results:Salvianolic acid B (5 μmol/L) could inhibit the proliferation of A549 and H1299 cells. Clone formation assay showed that Salvianolic acid B increased the radiosensitivity of A549 and H1299 cells, with a radiosensitization ratio of 1.45 and 1.38, respectively. Transwell chamber assay indicated that the ability of cell migration was significantly inhibited by Salvianolic acid B ( P<0.05). Western blot revealed that the expression levels of OTUD7B in A549 and H1299 cells were induced by irradiation in a time-dependent manner. Salvianolic acid B could down-regulate the expression levels of MMP-2, MMP-9 and p-Akt, whereas up-regulate the expression level of E-cadherin by down-regulating the expression level of OTUD7B. Conclusions:Salvianolic acid B can enhance the radiosensitivity of A549 and H1299 cells. The possible mechanism is that Salvianolic acid B down-regulates the expression level of OTUD7B induced by irradiation and inhibits the epithelial-mesenchymal transition process of tumor cells.

Effects of microRNAs on invasion, migration and apoptosis of lung cancer cell lines / 解剖学报

Objective To investigate the effects of microRNAs(miRNAs) on invasion, migration and apoptosis of lung cancer cell lines. Methods Target cells were infected with hsa-mir-933, hsa-mir-4700-3p, hsa-mir-3144-3p, hsamir-3972, and hsa-mir-548a-5p. Cell apoptosis was measured using flow cytomertry, Transwell invasion assay, cell migration were analyzed by lineation,miRNA was quantified using Real-time PCR. Results Compared with the vector and control, in hsa-mir-933, hsa-mir-4700-3p, hsa-mir-3144-3p, hsa-mir-397 and hsa-mir-548a-5p group the apoptosis significantly increased,cell invasion ability significantly decreased,cell migration ability significantly decreased, miRNAs expression significantly increased. Conclusion Five microRNAs can promote the apoptosis of lung cancer cell lines, change the high invasiveness of lung cancer cell lines, inhibit the migration of lung cancer cell lines, and increase the expression of corresponding miRNAs.

Study on Synthesis and Anti-tumor Activity of 6 Kinds of Tectorigenin Mannich Base Derivatives / 中国药房

OBJECTIVE： To conduct structural modification of tectorigenin to search for new compounds with anti-tumor activity. METHODS： Tectorigenin was used as a lead compound， and then added into amine reagents as ethanolamine， methylamine， ethylamine， dimethylamine， diethylamine， n-propylamine and formaldehyde solution. Tectorigenin Mannich base derivatives were synthesized by mannich reaction with as the lead compound. The structures of the derivatives were identified according to IR， UV， MS and NMR data. Solubility of tectorigenin and its derivatives were investigated by solubility test method. MTT assay was used to investigate the inhibitory effects of tectorigenin and its derivatives on the proliferation of human colon cancer cell line HCT116， human lung cancer cell line A549 and human hepatoma cell line HepG2， and half inhibitory concentration （IC50） was calculated. The inhibition rate of tectorigenin and its derivatives （100 mg/kg） on H22 hepatoma-bearing mice in vivo was studied. RESULTS： Totally of 6 kinds of tectorigenin mannich base derivatives were synthesized， such as 8-（N-hydroxyethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-methyl）-methyleneamino-5，7，4′-trihydroxy-6- methoxyisoflavone， 8-（N， N-diethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N， N-dimethyl）-methyleneamino- 5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-ethyl）-methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone， 8-（N-propyl）- methyleneamino-5，7，4′-trihydroxy-6-methoxyisoflavone （compounds 1-6 in turn）. Compared with tectorigenin， the water solubility of six derivatives was significantly improved， and the solubility was 5-20 times higher than that of tectorigenin. IC50 of compounds 1， 3 and 5 to HCT116 cells were （34.82±3.27）， （16.21±4.13）， （33.12±3.25） μmol/L， which were stronger than that of tectorigenin [（45.23±5.74） μmol/L]； IC50 of compounds 1， 3 and 5 to A549 cells were （37.05±5.74）， （26.88±4.52）， （30.13±6.23） μmol/L， which were stronger than that of tectorigenin [（53.24±6.34） μmol/L]； IC50 of compounds 1， 3 and 5 to HepG2 cells were （23.74±1.45）， （18.96±2.34）， （30.95±2.87） μmol/L， which were stronger than that of tectorigenin [（48.98±2.58） μmol/L]. Compounds 1， 3 and 5 showed higher inhibition rates （55.51％， 57.20％ and 49.15％） than tectorigenin （33.05％） on H22 hepatoma-bearing mice， respectively. The other three compounds had no obvious advantage over tectorigenin in anti-tumor activity. CONCLUSIONS： In this study， compounds 1， 3 and 5 of six tectorigenin mannich base derivatives synthesized in this study have stronger antitumor activity than tectorigenin.

Silencing LncRNA HOXD-AS1 enhances the radiosensitivity of H460 cells by regulating miR-454-3p expression / 中华放射肿瘤学杂志

Objective To explore the effect of LncRNA HOXD-AS1 on the radiosensitivity of H460 cells by targeting the regulation of miR-454-3p expression. Methods H460 cells were irradiated with 0, 2, 4, 6 and 8 Gy, and the expression levels of HOXD-AS1 and miR-454-3p were detected by qRT-PCR. Cell apoptosis rate were measured by flow cytometry experiments. Cell cloning experiments were used to detect cell radiosensitivity. The protein expression levels of Caspase-3, Cyclin D1 and γ-H2AX were detected by Western blot. Results The expression of HOXD-AS1 in H460 cells was significantly increased after X-ray irradiation ( P<0.05) , while the expression of miR-454-3p was significantly decreased ( P<0.05) . Silencing HOXD-AS1 significantly promoted apoptosis, increased radiosensitivity of lung cancer cells, promoted Caspase-3, γ-H2AX protein expression, and inhibited Cyclin D1 expression. HOXD-AS1 could target the expression of miR-454-3p. Conclusion Silencing HOXD-AS1 can promote the apoptosis of lung cancer cells and inhibit cell survival by targeting miR-454-3p to increase the radiosensitivity of lung cancer cells.

Liver kinase B1 gene enhances radiosensitivity of lung cancer H460 cells:an in vivo study / 中华放射肿瘤学杂志

Objective To investigate the effect of liver kinase B1(LKB1) on the radiosensitivity of subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.Methods Human lung cancer H460 cells were implanted into female nude mice (BALB/c-nu) to establish a subcutaneous xenograft tumor model of lung cancer.A total of 24 female nude mice in which the model was successfully established were equally and randomly divided into four groups:pEGFP-Ctrl plasmid (empty vector plasmid) group, irradiation (IR)+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid (overexpressing LKB1) group, and IR+pEGFP-LKB1 plasmid group.The growth of xenograft tumors was observed and the tumor inhibition rate and enhancement factor (EF) were calculated.The expression of LKB1 in each group was measured by immunohistochemistry and Western blot to analyze the relationship between LKB1 and radiosensitivity.Results Compared with the pEGFP-Ctrl plasmid group, the IR+pEGFP-Ctrl plasmid group, pEGFP-LKB1 plasmid group, and IR+pEGFP-LKB1 plasmid group showed varying degrees of inhibition of tumor growth, particularly in the IR+pEGFP-LKB1 plasmid group, and the tumor inhibition rates were 31.30%, 14.78%, and 43.48%, respectively.The EF of LKB1 in the IR+pEGFP-LKB1 plasmid group was 1.18.The immunohistochemistry and Western blot showed that LKB1 could be effectively expressed in the pEGFP-LKB1 plasmid group and IR+pEGFP-LKB1 plasmid group, but not in the other two groups.Conclusions The subcutaneous xenograft tumor model of human lung cancer H460 cells has been successfully established in nude mice.LKB1 has a radiosensitizing effect on the subcutaneous xenograft tumor of lung cancer H460 cells in nude mice.

Effects of bufalin combined with doxorubicin on the proliferation and apoptosis of human lung cancer cell line A549 in vitro / 中南大学学报(医学版)

Objective:To explore the effects ofbufalin (BUF) combined with doxorubicin (DOX) on the proliferation and apoptosis in human lung cancer cell line A549 in vitro.Methods:Methyl thiazolyl tetrazolium (MTT) assay was used to measure the inhibitory effects of BUF,DOX and their combination on the growth ofA549 cells.Hoechst 33342 staining was used to observe the changes of nucleus.Flow cytometry was used to investigate the apoptosis and cell cycle distribution of A549 cells.Western blot was used to examine the expression of apoptotic protein.Results:BUF and DOX showed inhibitory effect on the A549 cells in a dose and time-dependent manner.Compared with BUF or DOX alone,combination of BUF (1,20,100 nmol/L) with DOX (1.0 μg/mL) could significantly increase the growth inhibition rate ofA549 cells at 24,36,72 h,respectively (all P＜0.05).BUF and DOX alone could induce apoptosis,and their combination could significantly increase the apoptosis ratio.In addition,BUF combined with DOX could block the cell stage of A549 cells,keep the cell stage stay in S stage and up-regulate the expression of caspase-3.Conclusion:BUF combined with DOX can significantly inhibit the proliferation ofA549 cells,which might be related to the induction of apoptosis,cell cycle S phase arrest and caspase-3 up-regulation.

Antioxidant and cytotoxic activity of Tecoma stans against lung cancer cell line (A549)

ABSTRACT Human have been constantly using plants and plant products to overcome many diseases. The antioxidant property of the plant sources is studied to obtain an efficacious drug against cancer. The objectives of the present study is to evaluate the antioxidant and cytotoxic activity of the Tecoma stans extracts against lung cancer cell line in comparison with vincristine drug. The antioxidant activity was studied using the standard DPPH assay and the cytotoxic activity using MTT assay. DPPH assay results show that methanolic extract of T. stans in higher concentration show better antioxidant potential than the standard L-ascorbic acid. They exhibited strong antioxidant potential at 20 µg/mL concentration. The absorbance at 517 nm showed that in the range of 0.201-0.0203 compared to that of absorbance of ascorbic acid at 0.023.Cytotoxic activity was studied using MTT assay which showed that the increase in concentration of extract increases the cell death. At 100µg/mL concentration there is an increased cytotoxic activity, i.e., 99% of cell inhibition. The results of antioxidant and anticancerous activity may be positively correlated.

Rapid synthesis of biocompatible silver nanoparticles using aqueous extract of Rosa damascena petals and evaluation of their anticancer activity

OBJECTIVE@#To optimize the process parameters involved in the green synthesis of silver nanoparticles (G-SNPs) by aqueous extract of Rosa damascena petals and to evaluate the biocompatibility and anti cancer activity of the synthesized silver nanoparticles against human lung adenocarcinoma (A549).@*METHODS@#The process variables that include concentration of extract, mixing ratio of reactants, silver salt concentration and interaction time were analyzed. The compatibility of the G-SNPs was verified by incubating with erythrocytes and the anticancer property of the G-SNPs against A549 cells was performed by MTT assay.@*RESULTS@#Formation of G-SNPs was confirmed by the visual change in the colour of the reaction mixture from pale yellow to brown yellow. Surface plasmon resonance of synthesized G-SNPs was observed at 420 nm; the size of G-SNPs were analyzed by DLS and found to be in the range of (84.00±10.08) nm. Field emission scanning electron microscope and high resolution transmission electron microscopy analysis confirmed that the G-SNPs were fairly spherical. Fourier transform infrared spectroscopy spectroscopy and X-ray diffraction revealed the characteristic peaks of G-SNPs. Energy dispersive X-ray analysis showed a signal of silver around 3 keV. The synthesized G-SNPs exhibited anticancer activity as evidenced by the MTT assay. IC50 value of G-SNPs was found to be 80 μg/mL.@*CONCLUSION@#The results of the present study suggest that G-SNPs can be synthesized rapidly within first minute of the reaction; they are biocompatible and possess anticancer activity against human lung adenocarcinoma.

Rapid synthesis of biocompatible silver nanoparticles using aqueous extract of Rosa damascena petals and evaluation of their anticancer activity

Objective: To optimize the process parameters involved in the green synthesis of silver nanoparticles (G-SNPs) by aqueous extract of Rosa damascena petals and to evaluate the biocompatibility and anti cancer activity of the synthesized silver nanoparticles against human lung adenocarcinoma (A549). Methods: The process variables that include concentration of extract, mixing ratio of reactants, silver salt concentration and interaction time were analyzed. The compatibility of the G-SNPs was verified by incubating with erythrocytes and the anticancer property of the G-SNPs against A549 cells was performed by MTT assay. Results: Formation of G-SNPs was confirmed by the visual change in the colour of the reaction mixture from pale yellow to brown yellow. Surface plasmon resonance of synthesized G-SNPs was observed at 420 nm; the size of G-SNPs were analyzed by DLS and found to be in the range of (84.00±10.08) nm. Field emission scanning electron microscope and high resolution transmission electron microscopy analysis confirmed that the G-SNPs were fairly spherical. Fourier transform infrared spectroscopy spectroscopy and X-ray diffraction revealed the characteristic peaks of G-SNPs. Energy dispersive X-ray analysis showed a signal of silver around 3 keV. The synthesized G-SNPs exhibited anticancer activity as evidenced by the MTT assay. IC

Effects of genistein on proliferation and apoptosis of human non-small cell lung cancer cell line A549/DDP / 中国免疫学杂志

Objective:To observe the effects of genistein on proliferation and apoptosis of human non -small cell lung cancer cell line A549/DDP.Methods:①MTT assay was applied to evaluate the resistance index of A 549/DDP cell line to cisplatin and half in-hibitory concentration ( IC50 ) .②Inhibition rate of A549/DDP cell proliferation and IC 50 value were evaluated by MTT assay after treat-ment with 0, 1.25, 2.5, 5.0, 10, 20, 40, 60, 80 μg/ml genistein for 48 hour respectively.③A549/DDP cell cycle and apoptosis were evaluated by flow cytometry after treatment with 6.25, 12.5, 25 μg/ml genistein for 24 hours respectively.Results:①In expo-sing to cisplatin, the IC50 of A549 and A549/DDP was 33.6 μmol/L and 76.9 μmol/L respectively.The resistance index was 2.3. Cell growth inhibition rate increased following the cisplatin concentration increasing gradually .②A549/DDP growth inhibition rate in-creased at first and later decreased gradually following treatment with the genistein dose increased .The IC50 of A549 and A549/DDP was about 85.1 μg/ml and 80.2μg/ml respectively.③After treatment with 6.25, 12.5, 25μg/ml genistein for 24 hours, there were more A549/DDP cells arresting and showing apoptosis along with the genistein dose increased .Conclusion: Genistein can inhibit A549/DDP proliferation, cause A549/DDP arresting in G2/M phase and induce A549/DDP cell apoptosis with dose dependently .

Inhibitory effect of epigallocatechin gallate on the proliferation of Lewis Lung Cancer mice and A549 and Calu-3 lung cancer cells / 中国药学杂志

OBJECTIVE: To determine the inhibitory effect of EGCG on the Lewis Lung Cancer Model, to compare and preliminary to discuss the mechanism of the inhibitory effect of EGCG on the proliferation of A549 cell line and Calu-3 cell line. METHODS: Observed the inhibitory effect of EGCG on Lewis Lung Cancer in vivo. Compared the inhibitory effect of different dosages of EGCG on Calu-3 and A549 cell lines with WST-8 assay. Observed the opoptosis promotional effect of EGCG on Calu-3 cells. Compared the inhibitory effect of different dosage of EGCG on Calu-3 and A549 cell lines in PI cell number counting assay. RESULTS: EGCG inhibited the proliferation of Lewis Lung Cancer in vivo. Results of WST-8 assay showed that EGCG inhibited the proliferation of Calu-3 cell line in dose-dependent manner. However, A549 cell line showed sign of drug-resistance to EGCG. The inhibitory effect of EGCG on the proliferation of Calu-3 cell line through increasing apoptosis was observed with TUNEL assay. Cell number of Calu-3 was obviously decreased by treating of EGCG, but was not changed in A549 cell line. CONCLUSION: EGCG inhibited proliferation of Lewis Lung Cacer in vivo, and inhibited proliferation of Calu-3 cell line but didn't play the inhibitory role on A549 cell line in vitro. Copyright 2012 by the Chinese Pharmaceutical Association.

Effect of Hydroxycamptothecin on Proliferation of Human Lung Cancer Cell Line A549 / 中国中医药信息杂志

Objective To observe the effect of Hydroxycamptothecin (HCPT) on proliferation of human lung cancer cell line A549 in vitro. Method Using cell calture and MTT assay to observe the effect of HCPT on proliferation of human lung cancer cell line A549. Result Lower concentration of HCPT had no evident inhibitory effect on the human lung cancer cell line A549 after 24 h. The effect was evident when the concentration of HCPT was up to 50 ?g/mL, and the inhibitory rate was 44.17%. The inhibitory rate was 50.28% when the concentration of HCPT was up to 100 ?g/mL. The inhibitory effect of HCPT became more significant with the stimulation of the time, and the inhibitory rate of 100 ?g/mL concentration of HCPT was 70.98% after 48 h. Conclusions HCPT can inhibit the proliferation of human lung cancer cell A549 in vitro. The effect is dose and time dependent.

Inducing Apoptosis of NCI-H157 Human Lung Carcinoma Cells via Activation of Caspase Cascade by Combination Treatment with Arsenic Trioxide and Sulindac / 결핵

Arsenic trioxide(As2O3) was introduced into the treatment of refractory or relapsed acute promyelocytic leukemia. Some investigators have reported that arsenic trioxide had induced apoptosis in a variety of solid human tumor cell lines, including non-small cell lung cancer. Non-steroidal anti-inflammatory drugs(NSAIDs) are powerful chemopreventive agents for gastrointestinal cancers and the growth of established tumors are reduced by inducing apoptosis. It's also reported that NSAIDs enhanced tumor response to chemotherapeutic drugs or radiation. In this study, we aimed to determine whether combination of arsenic trioxide with sulindac augmented its apoptotic potential in NCI-H157 human lung cancer cells. The human lung cancer cell line NCI-H157 was treated with arsenic trioxide and sulindac. Cell viability was measured by the MTT assay. Apoptosis was measured by nuclear staining and flow cytometric analysis. The catalytic activity of the caspase families were measured by the fluorogenic cleavage of biosubstrates. The western blotting were also performed to define the mechanical basis of apoptosis. Combination treatment of arsenic trioxide and sulindac decreased the viability of NCI-H157 human lung cancer cells in a dose-dependent manner. The catalytic activity of caspase-3, 8 and 9 proteases were increased after combination treatment. Consistently PARP was cleaved from 116kDa to 85kDa fragments, and the expression of ICAD was decreased by time-dependent manner. Also combination treatment increased the expression of Fas and Fas/L. Combination therapy of arsenic trioxide with sulindac augments cell death and induces apoptosis via the activation of caspase cascade in NCI-H157 human lung carcinoma cells.

Lowly expression of miR-223 in CXCR4 positive cells from Lewis lung carcinoma cell line and its target gene prediction / 第三军医大学学报

Objective To analyze the differential expressions of miR-146a,miR-206,miR-223 and let-7c-1,such as cell differentiation-related miRNAs,in CXCR4-positive and CXCR4-negative subsets of the Lewis lung cancer cell lines(LLC).Methods CXCR4-positive and CXCR4-negative subsets were isolated from LLC by immunomagnetic beads sorting,and then their total cellular RNA were extracted by Trizol,expression of 4 miRNAs were detected by real-time fluorescence quantitative PCR(TaqMan probe),and potential target genes of miRNA whose differential expression was the most significant were predicted.Immunohistochemistry was carried out to confirm differential expression of the key molecule of certain research value within CXCR4-positive and CXCR4-negative subsets growing tumor tissue,and a BLAST search was performed to identify homologies of its 3′UTR.Results Compared to CXCR4-negative subsets,the expression of 4 miRNAs were lower in CXCR4-positive subsets,and expression of miR-223 had the most significant difference(Fold change=8.26).By softwares forecasting,miR-223 had potential target sites of IGF1R,IGFBP5,Pik3cb,ELK-1 and E2F1 mRNA,such as key molecular of IGF1R signaling pathway.The expression of IGF1R of CXCR4-positive subsets growing tumor tissue was significantly higher than that of CXCR4-negative subsets.Conclusion miR-223 is lowly expressed in CXCR4 positive cells from Lewis lung carcinoma cell lines.Position 238～244 nt and 688～695 nt in target sequences of 3′UTR of IGF1R mRNA was highly homologous by screening.Close correlation is found between miR-223 and IGF1R signaling pathway.The mechanisms underlying this biologically important finding need to be further explored.

Effects of Zhiling capsule on apoptosis and expressions of hTERT,CD44 in small cell lung cancer cell line NCI-H446 / 中国药理学通报

Aim To explore the mechanisms of the apoptosis induction and the effects of adhesion suppression of Zhiling capsule (ZLJN) in small cell lung cancer cell line NCI-H446.Methods According to the different components of ZLJN,NCI-H446 cells were treated with traditional Chinese medicine,western medicine and ZLJN composite groups.Apoptotic cells were tested by light microscopy,Hochest33258 staining method.The mRNA and protein expressions of bcl-2,bax and hTERT were analyzed by RT-PCR and Western blot respectively.The expressions of CD44 were detected by flow cytometry.Results After NCI-H446 cells were treated with different drug groups,The morphological changes of apoptotic cells were found by light microscopy and Hochest33258 staining method.The mRNA and protein expressions of bcl-2 were down-regulated while the expressions of bax were up-regulated compared to the control groups(P

Expression of vWF and integrin?_3 in H460 cells and association between the two factors and tumor metastasis / 中国癌症杂志

Background and purpose:The impact of the two factors of von Willnbrang factor(vWF) and integrin?3 on tumor metastasis has not been recognized. This study was done to investigate the expression of vWF and integrin?3 in human lung cancer cell line H460,the association between the two factors and tumor metastasis effect of vWF and integrin?3 on adhesion of human lung cancer cell line H460. Methods:The expression of vWF and integrin ?3 protein was examined by immunohistochemical staining. The impact of vWF and integrin?3 on adherent effect of tumor cells was evaluated by adhesion experiment, antibody inhibiting experiment and MTT.Results:Positive expression of vWF and integrin?3 was detected in H460 human lung cancer cells. H460 human lung cancer cells were able to adhere to vWF. Using anti-integrin?3, we observed that the ability of cell adhesion to vWF was inhibited,and A Value decreased from 1.59?0.06 to 0.55?0.03(P=0.01619). Using anti-vWF, we observed cell adhesion to vWF was inhibited too and A Value decreased from 1.60?0.06 to 0.54?0.03(P=0.01598),which had the same effect when using anti-integrin?3.Conclusions:H460 human lung cancer cells are capable of producing vWF, and vWF expression contributes to metastasis by adhering to cancer cells, integrin?3 is the vWF receptor on H460 human lung cancer cells.

Establishment and Characterization of a Small Cell Lung Cancer Cell Line(JePa-1)

Lung cancer is the most common malignant tumor worldwide and its incidence continues to rise each year. Recent development of molecular biologic method has led to advances in determining the etiologic factors of lung cancer and the establishment of cell lines has provided a lot of information on the through chemosensitivity, radiation biology studies, cytogenetics, and molecular biologic studies, which permits improved treatment for lung cancer. We established a small cell lung cancer cell line, designated JePa-1, obtained from malignant pericardial effusion of small cell lung cancer patient and characterized its morphologic and molecular biologic features. the JePa-1 cell line grew relatively slowly (doubling time 45hrs) as very loosely adherent floating aggregates growing in small clumps with distinct cell outlines and intertwined cords. Also JePa-1 cell line secreted antidiuretic hormones. Electronmicroscopic examination revealed that JePa-1 cell line and xenografts contained electron dense core granules, characteristic of being of neuroendocrine origin. To investigate the tumorigenic capacity, the JePa-1 cell line was injected into SCID and nude mice. Tumors taken from xenografts were observed in 3 out of 4 of the SCID mice and 2 out of 4 of the nude mice. The histologic characteristics of the xenografts were similar to those of the cell line and the original cytologic finding of the pericardial fluid, suggesting small cell carcinoma. The results of immunohistochemical markers showed reactivity for Rb protein, c-myc, TGF-alpha, TGF-beta , EGFR, keratin, NSE, chromogranin, and EMA. The DNA ploidy and the index of the JePa-1 cells was tetraploid and 2.13, respectively. The positive rate for the Rb, c-myc and K-ras proteins of the JePa-1 cell line were 98.9%, 99.3%, and 99.7% respectively as determined by flow cytometry. Cytogenetic analysis using the G-banding technique showed 65 chromosomes with various numerical and structural abnormalities. On examination of the expression of TGF-alpha, TGF-beta , and EGFR by PCR, only the EGFR was positive Through the establishment of JePa-1 cell line, we report in this paper the characterization of a small cell lung cancer such as morphologic and immunocytochemical features, growth characteristics in culture, hormone production, expression of oncoprotein and several growth factors, tumorigenicity, chromosomal abnormalities, and DNA ploidy and index. The JePa-1 cell line will be valuable in vitro studies for the etiology, treatment and the prognostic factors in small cell lung cancer.