ABSTRACT
OBJECTIVE：To prepare cell penetrating peptide PFV-modified paclitaxel （PTX）/artesunate（ART）co-loaded targeting micelles ，and to investigate in vitro anti-tumor activity. METHODS ：According to optimal technology ，PFV-modified PTX/ ART co-loaded targeting micelles were prepared by membrane hydration method ，and were characterized. Using blank micelle as blank control ，sulforhodamine B （SRB）method was used to evaluate the toxicity of PTX micelles ，ART micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to human gastric cancer BGC- 823 cells. The coumarin was used as fluorescent probe replacing PTX to prepare corresponding micelles. Then ，the uptake of BGC- 823 cells to corresponding micelles and targeting effect were observed and determined by flow cytometry and fluorescence microscope. The effects of PTX micelles ， ART micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles on the invasion of BGC- 823 cells were investigated by Transwell chamber method. RESULTS ：Average particle size of PFV-modified PTX/ART co-loaded targeting micelles was （51.30±3.95）nm；PDI was 0.19±0.01，and Zeta potential was （0.21±0.02）mV. The encapsulation efficiency of PTX and ART were higher than 90％. The shape of micelles were spherical. The blank micelles had no obvious toxicity to BGC-823 cells. The IC 50 value of PTX micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to BGC-823 cells were （3.09±0.22），（1.93±0.24），（1.11±0.15）μmol/L，respectively. The distribution amount of different micelles in BGC- 823 cell nucleus in the descending order were PFV-modified coumarin/ART micelles ＞coumarin/ART micelles ＞coumarin micelles＞blank control. The order of inhibitory effect was PFV-modified PTX/ART co-loaded targeting micelles ＞PTX/ART micelles＞ART micelles ＞PTX micelles ＞blank control. CONCLUSIONS： Prepared PFV-modified PTX/ART No.81874347） co-loaded targeting micelles are in line with the quality of 1915286446@qq.com Chinese Pharmacopoeia . It shows strong cytotoxicity to BGC-823 cells，can improve the drug targeting and the cell uptake，and inhibit the inv asion and metastasis of BGC- 823 cells.
ABSTRACT
Objective: To investigate the effects of overexpression of microspherule protein 1 (MCRS1) in the gastric cancer cells on the invasion and migration of gastric cancer cells, and to elucidate their possible mechanisms. Methods: The gastric cancer BGC-823 cells, SGC-7901 cells and the normal gastric mucosal epithelial GES-1 cells were cultivated. Western blotting method was used to detect the expressions of MCRS1 in three kinds of cells. The result showed that MCRS1 protein had the lowest expression in the BGC-823 cells, so the gastric cancer BGC-823 cells were selected for next experiments. The recombinant plasmid of MCRS1 was constructed, and the gastric cancer BGC-823 cells in the logarithmic growth phase were selected. Blank group, empty vector transfection group and MCRS1 transfection group were established, and the plasmid was transfected into the BGC-823 cells using Lipo3000. The expression levels of epithelial cadherin (E-cadherin) protein, neuronal cadherin (N-cadherin) protein and Snail protein were detected by Western blotting method. The cell scratch assay and Transwell assay were used to detect the migration and invasion of gastric cancer BGC-823 cells. Results: Compared with the normal gastric epithelial GES-1 cells, the expression level of MCRS1 protein in the gastric cancer BGC-823 cells was decreased (P < 0 . 01), but the expression level of MCRS1 protein in the SGC-7901 cells was increased (P < 0 . 01). The PCR identification and sequencing analysis showed that the MCRS1 recombinant plasmid was successfully constructed. Compared with blank group and empty vector transfection group, the expression levels of MCRS1 protein and E-cadherin protein in MCRS1 transfection group were increased (P
ABSTRACT
PURPOSE: Information on the possible role of the ribosomal protein S15a (RPS15a) in gastric cancer is scarce. The aim of this study was to evaluate the impact of RPS15a gene expression on the growth and cell cycle of gastric cancer cells in vitro and in vivo. MATERIALS AND METHODS: RPS15a mRNA expression was examined in cancer tissues and their corresponding adjacent normal tissues of 40 gastric adenocarcinoma patients. Next, RPS15a was knocked down using a lentivirus-mediated RNA interference (short hairpin RNA) system in the gastric cancer cell line BGC823. The effect of RPS15a knockdown was examined using CCK-8 assay, cell scratch test, colony formation assay, and flow cytometry. Finally, in nude mice, a tumorigenicity test was performed, and the tumor volume and weight were measured. RESULTS: RPS15a expression in tumor tissue was significantly greater than that in the adjacent normal tissue of gastric cancer patients. After RPS15a silencing, the BGC823 cell proliferation rate decreased significantly; most cells were arrested in the G0/G1 phase, cell growth was inhibited, and the migration rate was decreased. Colony formation assay showed that the number and size of clones in the RPS15a-silenced cells were fewer and smaller, compared to control cells. The nude mouse tumorigenicity test showed that RPS15a silencing had an inhibitory effect on tumor volume and mice weight. CONCLUSION: The present study found RPS15a expression to be higher in gastric tumors and its silencing in gastric cancer cells to inhibit the proliferation, growth, and migration thereof. Accordingly, RPS15a may be considered as a potential therapeutic target in gastric cancer.
Subject(s)
Animals , Humans , Mice , Adenocarcinoma , Carcinogenicity Tests , Cell Cycle , Cell Line , Cell Proliferation , Clone Cells , Flow Cytometry , Gene Expression , Gene Silencing , In Vitro Techniques , Mice, Nude , Ribosomal Proteins , RNA Interference , RNA, Messenger , Sincalide , Stomach Neoplasms , Tumor BurdenABSTRACT
Objective To investigate the effect of Wulong Xiaozheng Pill (WXP) on the migration and invasion of human gastric cancer cell-line BGC-823 and its mechanism. Methods WXP, IGF-1, and LY294002 were added on BGC-823 cells. Then the inhibitory effect of WXP was detected by MTT assay. Transwell assay was performed to determine the migration and invasion capacity of on BGC-823 cells. Expressions of VEGF, MMP-2, and MMP-9 were detected by ELISA, while the expressions of related proteins and mRNA in PI3K/NF-κB signaling pathway were detected by Western blotting and RT-PCR. Results WXP can inhibit the proliferation, adhesion, invasion, and migration of BGC-823 cells. In addition, WXP inhibited the expression of VEGF, MMP-2 and MMP-9 protein in BGC-823 cells. WXP significantly inhibited the expression of p-PI3K, p-Akt, p-IKK-a, and p-NF-κB p65Ser 276 proteins, PI3K, Akt, IKKa, and NF-κB mRNA, which showed a time-dependent and dose-dependent manner. Conclusion WXP inhibit the capacity, migration and invasion of BGC-823 cells by blocking PI3K/NF-κB signaling pathway.
ABSTRACT
AIM:To investigate the enhancing effect of quercetin on the 5-fluorouracil-induced apoptosis in gastric cancer.METHODS: MTT assay was conducted to evaluate the effect of quercetin on the 5-fluorouracil-induced death of gastric cancer cell line BGC-823.Co-immunoprecipitation and Western blot analysis were performed to detect the expression of c-Jun and Bcl-xL,phosphorylation of c-Jun,activation of caspase-9 and caspase-3,and release of cytochrome C in BGC-823 cells treated with quercetin and 5-fluorouracil.The apoptosis of BGC-823 cells treated with quercetin and 5-fluorouracil was analyzed by flow cytometry.RESULTS: Adjuvant therapy of quercetin significantly enhanced the 5-fluorouracil-induced death of BGC-823 cells.Meanwhile, quercetin decreased the half maximal inhibitory concentration (IC50)of 5-fluorouracil to BGC-823 cells.Quercetin treatment significantly inhibited the expression of c-Jun,and inhibited the 5-fluorouracil-induced phosphorylation of c-Jun and the interaction between c-Jun and activating transcription factor 2 (ATF2).Subsequently,quercetin inhibited the up-regulation of Bcl-xL induced by 5-fluorouracil in the BGC-823 cells. However,transfection with c-Jun plasmid abolished the promoting effect of quercetin on 5-fluorouracil-induced cell death. In addition, quercetin promoted 5-fluorouracil-induced release of cytochrome C from mitochondria and caspase-dependent apoptosis in BGC-823 cells.CONCLUSION:Quercetin treatment enhances 5-fluorouracil-induced mitochondrial apoptosis in BGC-823 cells through c-Jun/ATF2/Bcl-xL pathway.
ABSTRACT
Objective To study the effect of annonaceous acetogenins (ACGs) on human gastric cancer cells in vitro. Methods After ACGs were administered to gastric cancer cells in vitro, the cell viability, cell adhesion ability and cell migration ability were assessed by MTT assay, adhesion assay and wound-healing assay, respectively. Results ACGs inhibited the cell viability, adhesion ability and migration ability in a dose-dependent manner in gastric cancer cells. Conclusion ACGs could inhibit cell activities of human gastric cancer cells in viro, and will be developed as a promising anticancer candidate and used in gastric cancer.
ABSTRACT
AIM:To investigate the effect of dihydroartemisinin ( DHA) adjuvant treatment on enhancing the antitumor effect of 5-fluorouracil (5-FU) against gastric cancer .METHODS:The gastric cancer BGC-823 cells were di-vided into control group , DHA group, 5-FU group, 5-FU+DHA group and 5-FU+DHA+SIRT1 plasmid group.The via-bility of BGC-823 cells treated with DHA and 5-FU was measured by MTT assay .The expression of SIRT1 and NADPH ox-idase, activation of caspase-9 and caspase-3, and phosphorylation of ASK1 and JNK in the BGC-823 cells treated with DHA and 5-FU were determined by Western blot .The production of ROS and the apoptosis of the BGC-823 cells treated with DHA and 5-FU were analyzed by flow cytometry .RESULTS:Dihydroartemisinin significantly inhibited the expression of SIRT1 and increased NADPH oxidase protein level (P<0.05).DHA increased the sensitivity of BGC-823 cells to 5-FU, thus decreasing the IC50 of 5-FU to the gastric cancer cells.However, transfection with SIRT1 plasmid decreased the cytotoxicity of DHA and 5-FU co-treatment to the BGC-823 cells.DHA promoted the production of ROS and phosphoryla-tion of ASK1 and JNK induced by 5-FU in the BGC-823 cells ( P<0.05 ) .However , ROS scavenger N-acetylcysteine ( NAC) or JNK specific inhibitor SP600125 inhibited the cell death and activation of caspase-9 and caspase-3 induced by DHA and 5-FU co-treatment (P<0.05).In addition, NAC significantly inhibited the phosphorylation of JNK in the BGC-823 cells co-treated with DHA and 5-FU.However, treatment with SP600125 did not influence the ROS production in the BGC-823 cells, indicating that JNK was the downstream target of ROS pathway .CONCLUSION: Combination of DHA with 5-FU induces caspase-dependent apoptosis in gastric cancer cells through the SIRT 1/NADPH oxidase/ROS/JNK sig-naling pathway .
ABSTRACT
The aim of the study was to investigate the anti-proliferation and apoptosis-inducing effects of S1, a novel tetrandrine derivative, in human gastric cancer BGC-823 cells and explore the possible mechanism of action. The anti-proliferative activity was determined by MTT assay; the induction of cell cycle arrest and apoptosis were detected by flow cytometry. Quantitative real time RT-PCR and Western blotting were used to evaluate the mRNA and protein expression levels in mitochondrial pathway. S1 significantly reduced cell viability and induced a G2/M phase arrest and apoptosis in dose- and time-dependent manner. Further studies showed that S1 increased mRNA and protein expression of Bax and the Bax/Bcl-2 ratio. Moreover, S1 decreased the protein expression of procaspase-9 and procaspase-3, suggesting that the induction of apoptosis may be related to the alteration of the ratio of Bax/Bcl-2 and the activation of caspases. These findings suggested that S1 merits further investigation as a novel therapeutic agent for the treatment of human gastric cancer.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Apoptosis , Benzylisoquinolines , Chemistry , Pharmacology , Caspase 3 , Genetics , Metabolism , Caspase 9 , Genetics , Metabolism , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Cell Survival , Proto-Oncogene Proteins c-bcl-2 , Genetics , Metabolism , Stomach Neoplasms , Drug Therapy , Genetics , bcl-2-Associated X Protein , Genetics , MetabolismABSTRACT
Objective:To investigate the anti-proliferation effect of carvacrol on human gastric cancer cell BGC-823 and explore the molecular mechanism. Methods:The proliferation of BGC-823 cells was evaluated by MTT assay. Flow cytometry was used to ana-lyze the cell apoptosis after exposed to carvacrol. Transwell assay was used to analyze the effect of carvacrol on cell metastasis. Quanti-tative realtime-PCR was used to detect the expression of MMP-9 and TIMP-1. The expression of caspase-9 and PARP,and the activa-tion of ERK and P38 were detected by Western blot. Results:The incubation with carvacrol resulted in a significant inhibition of BGC-823 cell proliferation. After the treatment with carvacrol,the apoptotic rate was significantly increased( 0 μmol · L-1 vs 10 μmol · L-1 ,P<0. 000 1;0 μmol·L-1 vs 20 μmol·L-1 ,P<0. 000 1;0 μmol·L-1 vs 40 μmol·L-1 ,P<0. 000 1;0 μmol·L-1 vs 80μmol·L-1 ,P<0. 000 1)and the invasion ability was decreased(0 μmol·L-1 vs 80 μmol·L-1 ,P<0. 000 1). The expression of caspase-9(0 μmol·L-1 vs 80 μmol·L-1,P<0. 000 1)and TIMP-1 was increased(P<0. 000 1),PRAP fragment occurred(P<0. 000 1)and P38 signaling pathway was activated in the carvacrol treated group,while the expression of MMP-9 activity of ERK signa-ling pathway was inhibited(P<0.000 1). Conclusion:Carvacrol can inhibit cell growth and invasion,and induce cell apoptosis, which is closely related to MAPK signaling pathway.
ABSTRACT
Ziyuglycoside II is an active compound of Sanguisorba officinalis L. that has anti-inflammation, antioxidation, antibiosis, and homeostasis properties. We report here on the anticancer effect of ziyuglycoside II on human gastric carcinoma BGC-823 cells. We investigated the effects of ziyuglycoside II on cell growth, cell cycle, and cell apoptosis of this cell line. Our results revealed that ziyuglycoside II could inhibit the proliferation of BGC-823 cells by inducing apoptosis but not cell cycle arrest, which was associated with regulation of Bax/Bcl-2 expression, and activation of the caspase-3 pathway. Our study is the first to report the antitumor potential of ziyuglycoside II in BGC-823 gastric cancer cells. Ziyuglycoside II may become a potential therapeutic agent against gastric cancer in the future.
Subject(s)
Humans , Apoptosis/drug effects , /metabolism , /metabolism , Saponins/pharmacology , Signal Transduction/drug effects , /metabolism , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Carcinoma/drug therapy , /drug effects , Caspase Inhibitors/metabolism , Cell Cycle/drug effects , Cell Proliferation/drug effects , Fluorometry , Fluorouracil/pharmacology , /drug effects , Sanguisorba/chemistry , Stomach Neoplasms/drug therapy , /drug effectsABSTRACT
AIM: To observe the inhibitory effect of interferon-α ( IFN-α) on the growth invasiveness and metastasis of human gastric carcinoma cell line BGC-823, and mechanism of its action. METHODS: We detected the influence of IFN-α on the proliferative ability of BGC-823 in cell culture system, the cell vitality with the MTT colorimetric assay, and the cell cycle with flow cytometer (FCM). The regulatory functions of IFN-α to the expression of E-cadherin and matrix metalloproteinase-2 ( MMP-2) in tumor cells were estimated by immunohistochemical analysis ( S-P). The ultrastructural changes of the junction among the tumor cells were observed under electron microscope. RESULTS : IFN-α can significantly inhibit the growth of human gastric carcinoma cell line BGC-823 in a dose-dependent manner. When the concentration of IFN-α was ≥106 U/L, the cell proliferation can be effectively suppressed,the suppression rate was ≥ 12. 2%, and the blockage appeared at the phase of G1-S of the cell cycle. Under the induction of IFN-α, the expression level of the cell E-cadherin increased while the MMP-2 decreased. The changes on ultrastructure of the cells showed the increased adhesive junctions and the relative compact structure. CONCLUSION: IFN-α can suppress the growth of human gastric carcinoma cell line BGC-823 through its influence on cell cycle. IFN-α can regulate the expression of E-cadherin and MMP-2, make the cell junction closely, so that it has the potential on restricting the invasion and metastasis of gastric carcinoma cells.
ABSTRACT
AIM:To investigate the effect of tumor-suppressor RUNX3 on the transcription of apoptosis-related genes bcl-2,bax,caspase-3,caspase-8,caspase-9 in human gastric carcinoma cells BGC823,and to reveal the apoptosis molecular mechanism promoted by RUNX3. METHODS:The eukaryotic expression vector of human Runx3 gene pcDNA3.1-Runx3 was constructed. pcDNA3.1-Runx3 and blank vector pcDNA3.1 were transfected into BGC823 cells,respectively. After 48 h,the total mRNA and protein were acquired and the expression level of Runx3 was determined by RT-PCR and Western blotting. Then,the mRNA and protein expression of bcl-2,bax,caspase-3,caspase-8 and caspase-9 was determined by RT-PCR and Western blotting. ?-actin was used as a control. RESULTS:The eukaryotic expression vector pcDNA3.1-Runx3 was constructed successfully and transfected into BGC823 cells. RT-PCR and Western blotting confirmed that RUNX3 level was higher in pcDNA3.1-Runx3 transfected BGC823 cells than that in blank vector-transfected cells (P
ABSTRACT
AIM:To observe the inhibitory effect of interferon-?(IFN-?)on the growth invasiveness and metastasis of human gastric carcinoma cell line BGC-823,and mechanism of its action.METHODS:We detected the influence of IFN-? on the proliferative ability of BGC-823 in cell culture system,the cell vitality with the MTT colorimetric assay,and the cell cycle with flow cytometer(FCM).The regulatory functions of IFN-? to the expression of E-cadherin and matrix metalloproteinase-2(MMP-2)in tumor cells were estimated by immunohistochemical analysis(S-P).The ultrastructural changes of the junction among the tumor cells were observed under electron microscope.RESULTS:IFN-? can significantly inhibit the growth of human gastric carcinoma cell line BGC-823 in a dose-dependent manner.When the concentration of IFN-? was ≥106 U/L,the cell proliferation can be effectively suppressed,the suppression rate was ≥12.2%,and the blockage appeared at the phase of G_1-S of the cell cycle.Under the induction of IFN-?,the expression level of the cell E-cadherin increased while the MMP-2 decreased.The changes on ultrastructure of the cells showed the increased adhesive junctions and the relative compact structure.CONCLUSION:IFN-? can suppress the growth of human gastric carcinoma cell line BGC-823 through its influence on cell cycle.IFN-? can regulate the expression of E-cadherin and MMP-2,make the cell junction closely,so that it has the potential on restricting the invasion and metastasis of gastric carcinoma cells.
ABSTRACT
AIM To study the effect and mechanism of valdecoxib, a selective COX 2 inhibitor, on human gastric cancer BGC 823 cells. METHODS MTT assay and flow cytometry were used to observe the effect of valdecoxib on proliferation, apoptosis and the cell cycle distribution of BGC 823 cells. Laser confocal microscopy, transmission electron microscope and DNA fragmentation assay were further used to detect the apoptosis. The content of LDH was examined using LDH kit. RESULTS Valdecoxib in 25～400 ?mol?L -1 significantly inhibited the proliferation of BGC 823 cell in a time and dose dependent fashion, the inhibition rate of proliferation was 24 0%～92 0% after 72 h, and the rate of apoptosis was increased from (2 6?0 7)% to (7 6?1 5) %～(16 5?1 5)%. 100～400 ?mol?L -1 valdecoxib also decreased the proliferation index and the proportion of cells in the S phase, increased the proportion of cells in the G 0/G 1 phase, but had no effect on the proportion of cell in the G 2/M phase. CONCLUSION Valdecoxib inhibits human gastric cancer BGC 823 cells growth by inducing apoptosis and cell cycle arrest. The growth inhibitory effect of 400 ?mol?L -1 valdecoxib is also associated with cell necrosis.
ABSTRACT
Objective To study the role of protein kinase B(PKB) in gastric cancer BGC-823 cells treated by TPA. Methods Flow cytomety(FAC) and fluorescence microscope were used to detect the effect of TPA on gastric cancer BGC-823 cells,after cells were treated by BrdU and stained by DAPI,respectively.The effect of TPA on the expression level of PKB protein and its phosphorylation of Ser 473 and Thr 308 were investigated by Western blotting.When nuclear and cytoplasmic protein fractions were prepared through lysis of cell and centrifugation,the effect of TPA on the expression level of PKB protein and its phosphorylation of Ser 473 in cytoplasm and nuclei were detected by Western blotting.Meanwhile,the effect of TPA on the distribution of PKB was observed under laser-scanning confocal microscope with immunofluorescence technique. Results TPA induced the apoptosis of gastric cancer BGC-823 cells.It also induced the inhibition of PKB protein expression in a TPA concentration-dependent and time-dependent manner in gastric cancer BGC-823 cells,regardless of dephosphorylation of PP2A.TPA inhibited the phosphorylation of PKB at Ser 473,but it did not affect its phosphorylation at Thr308.TPA only attenuated the expression level of PKB and its phosphorylation of Ser 473 in nuclei,whereas it did not change the distribution of PKB in BGC-823 cells.Conclusion These data suggest that the inhibition of PKB plays an important role in mediating the effect of TPA in gastric cell line.Apoptosis induced by TPA is partly due to the inhibition of PKB and its phosphorylation of Ser 473 in nuclei.