RORα antagonist T0901317 promotes EMT in human gastric cancer MGC803 cells by RORα/β-catenin signal / 中国药理学通报

Aim To investigate the effect of RORα antagonist T0901317 promoting EMT (epithelial-mesenchymal transition) of human gastric cancer MGC803 cells by RORα/β-catenin signal. Methods Cell proliferation was detected by MTT. Cell migration and invasion were detected by cell scratch and Transwell assay respectively. RORα/β-catenin signaling molecules were detected by Western blot and immunofluorescence. RORα binding to β-catenin protein was detected by immunoprecipitation. Results MTT assay showed that the proliferation ability of T0901317 cells increased in a time-dependent manner compared with MGC803 cells (P < 0. 05). Cell scratches and Transwell experiments showed that the migration and invasion ability of T0901317 cells were significantly enhanced compared with MGC803 cells (P < 0. 05). Western blot analysis showed that RORα protein was significantly down-regulated after T0901317 compared with untreated group (P < 0. 05), and total β-catenin protein and nuclear β-catenin in MGC803 cells were up-regulated after T0901317 (P < 0. 05). Compared with the control group, RORα protein binding to β-catenin protein significantly decreased after T0901317 treatment (P < 0. 05). Compared with MGC803 cells treated with T0901317, the long spindle cells increased and the heteromorphism was more obvious. T0901317 significantly up-regulated the expression of Rac1, TGFβ1 and Vimentin in MGC803 cells (P < 0. 05), and inhibited the expression of E-cadherin (P < 0. 05). Conclusion T0901317 can promote the proliferation, migration, invasion and EMT in human gastric cancer MGC803 cells by RORα/β-catenin signal.

Extract from Modified Xiao Xianxiongtang Inhibits Epithelial-mesenchymal Transition and Invasion and Migration Mediated by TGF-β1 of Human Gastric Cancer MGC-803 Cells via Wnt5a/Ca2+/NFAT Signaling Pathway / 中国实验方剂学杂志

Objective:This studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition（EMT） and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁），and to explore the mechanism of regulating Wnt5a/Ca²⁺/ activated T-cell nuclear factor（NFAT） signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells. Method:TGF-<italic>β</italic>₁（10 μg·L^-1）was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment， scratchhealing experiment， Western blot and immunofluorescence assay were used to detect cell invasion and migration ability， expression of EMT marker protein and key protein expression of Wnt5a/Ca²⁺/NFAT signaling pathway， and intracellular Ca²⁺ concentration. Result:Compared with the blank group， TGF-<italic>β</italic>₁ could significantly enhance the invasion and migration ability of MGC-803 cells（<italic>P</italic><0.01）， down-regulate the level of E-cadherin（<italic>P</italic><0.01）， up-regulate protein expressions of N-cadherin， Snail and Vimentin（<italic>P</italic><0.01）， and induce cell Wnt5a， calcineurin （CaN）， total protein of activated T-cell nuclear factor 1（NFAT1）， up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca²⁺（<italic>P</italic><0.01）. Compared with the TGF-<italic>β</italic>₁ group， modified Xiao Xianxiongtang （10， 20， 40 mg·L^-1） could significantly inhibit this phenomenon，and 40 mg·L^-1 had the best effect（<italic>P</italic><0.05，<italic>P</italic><0.01）.The specific inhibitors of Wnt5a/Ca²⁺/NFAT signaling pathway （<italic>R</italic>）-（+）-Bay-K-8644 and modified Xiao Xianxiongtang （40 mg·L^-1） could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-<italic>β</italic>₁， up-regulate the level of E-cadherin， and down-regulate expressions of N-cadherin， Snail， Vimentin， Wnt5a， CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca²⁺（<italic>P</italic><0.05，<italic>P</italic><0.01）.Moreover， （<italic>R</italic>）-（+）-Bay-K-8644 combined with modified Xiao Xianxiongtang （40 mg·L^-1） had stronger inhibitory effect（<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:These results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-<italic>β</italic>₁ via Wnt5a/Ca²⁺/NFAT signaling pathway，thereby reducing the invasion and migration ability of MGC-803 cells.

The differential expression of microRNA in human gastric cancer MGC803 cells induced by diallyl disulfide / 中国药理学通报

Aim: To investigate the expression and i-dentification of differential miRNAs in human gastric cancer MGC803 cells induced by diallyl disulfide (DADS). Methods: Differential miRNAs expression in human gastric cancer MGC803 cells induced by DADS was detected and identified by miRNA chip and qPCR. Results: MiRNAs chip detection showed upregulation of miR-200b, miR-22, miR-7, miR-143, miR-138, miR-34a and miR-150, and down-regulation of miR-222, miR-21, miR-15b, miR-182 and miR-18a in differential miRNAs of MGC803 cells treated with 3 0 mg · IT-1 DADS at 24 h(P<0.05). And qPCR demonstrated that the expressions of miR-200b, miR-22, miR-7, miR-143, miR-138, miR-34a and miR-150 was up-regulated in MGC803 cells treated with 30 mg · L-1 DADS(P <0. 05). Moreover, qPCR showed that the expressions of miR-200b and miR-22 in various human gastric cancer cells including MGC803, BGC823, MKN28, SGC7901 and HGC27 cells were lower than normal human gastric cancer GES-1 cells (P <0. 05). The expression of miR-200b and miR-22 in gastric cancer tissues was significantly lower than that in normal gastric tissues (P < 0. 05). Conclusions: The expression of down-regulation of 7 miRNA and up-regulation of 5 miRNA in differential miRNAs in MGC803 cells induced by DADS. The expression of down-regulation of miR-200b and miR-22 in gastric cancer tissues and cells. DADS could up-regulate the expression of miR-200b and miR-22 in gastric cancer cells.

Rosmarinic acid derivative RAD-9 induces apoptosis in gastric cancer MGC-803 cells via PI3K/Akt and p38 MAPK signaling pathway / 中国药理学通报

Aim To study the apoptosis-inducing effect of rosmarinic acid derivative RAD-9 on gastric cancer MGC-803 cells and the underlying mechanisms.Methods MTT assay was taken to detect the survival of gastric cancer MGC-803 cells effected by RAD-9.Cell apoptosis was detected by flow cytometry.The apoptotic morphology of MGC-803 cells was observed by Hoechst 33258 staining.The protein expression levels of Bcl-2,Bax,caspase-3,Akt,p-Akt,p38 MAPK and p-p38 MAPK were measured by Western blot.Results The results of MTT assay showed that RAD-9 inhibited the viability of gastric cancer MGC-803 cells in a time and concentration-dependent manner.Flow cytometry showed that RAD-9 significantly promoted apoptotic cell percentage in gastric cancer MGC-803 cells (P ＜ 0.01).Hoechst 33258 staining showed that the nucleus of MGC-803 cells could be observed with typical apoptotic morphological changes after RAD-9 administration.Compared with the control group,the protein expression levels of Bcl-2,Akt,p-Akt were significantly down-regulated (P ＜ 0.01),while those of Bax,caspase-3,p38 MAPK,p-p38 MAPK were significantly up-regulated (P ＜ 0.01).Conclusion RAD-9 can inhibit the growth and further induce apoptosis in gastric cancer MGC-803 cells,which may involve inhibiting PI3K/Akt and activating p38 MAPK signaling pathway.

Induction of furanodiene on apoptosis of human gastric adenocarcinoma MGC-803 cells / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of furanodiene(FDE),a diterpene derived from the medicinal plant Zedoary,on apoptosis of human gastric cancer MGC-803 cells induced in vitro. METHODS MGC-803 cells were treated with FDE 46.29~740.74μmol·L-1 for 24,48 and 72 h,and the cell viability was detected with MTT assay. Cell morphology was observed by light microscopy and Hoechst33342 staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. Rh123 staining and fluorescence probe DCFH-DA were employed to detect the changes in mitochondrial membrane potential (MMP) and reactive oxygen species(ROS). RESULTS MTT Results showed that FDE 46.29-740.74μmol · L-1 exhibited significantly higher cytotoxicity to gastric cancer MGC-803 cells. IC50 for MGC-803 of 24,48 and 72 h treatment was 347.91,257.41 and 101.01μmol·L-1,respectively. Treatment with FDE 92.58-370.32μmol·L-1 for 24 h also caused significant morphological changes in MGC-803 cells. AnnexinⅤ-FITC/PI double staining showed that the apoptotic rate increased after FDE 92.58-370.32μmol·L-1 treatment for 24 h(P<0.05). FDE enabled MGC-803 cell cycle arrest in S phase. DCFH-DA staining showed that FDE resulted in an increase in intracellular ROS levels(P<0.05) when PDE concentration was 370.37μmol·L-1(P<0.05). MMP decreased after FDE treatment when PDE concen?tration was 370.37μmol·L-1(P<0.05). CONCLUSION FDE Possesses potent tumor selected toxicity and can induce apoptosis of MGC-803 cells through cell cycle arresting,which is related to inhibition of DNA biosynthesis.

Effects of combination treatment with L-carnitine and 5-fluorouracil on proliferation and apoptosis in MGC803 cells / 中国药学杂志

OBJECTIVE: To investigate the effects of combination treatment with L-carnitine and 5-fluorouracil on the proliferation and cell apoptosis of gastric cancer MGC803 cells. METHODS: MGC803 cells were divided into control group, 5-fluorouracil group and the combination of L-carnitine and 5-fluorouracil group (L-carnitine +/→5-fluorouracil group) in vitro. The inhibitory rate of cells was measured by MTT assay. The apoptosis rate and cell cycle of cells were detected by FLOW. Western blot was used to analyzed the expression of Bcl-2, Bax, adenine nucleotide translocator1 (ANT1) and cleaved-PARP. RESULTS: Compared with 5-fluorouracil group, the inhibition rate of MGC803 cells was increased when cells were treated with the combination of L-carnitine and 5-fluorouracil. The apoptosis rate of cells was raised and the cells were blocked at S phase. In addition, the combination group can decrease the expression of Bcl-2 and increase the expression of Bax, ANT1 and cleaved-PARP. At the same time, the apoptosis rate of cells and the cell cycle were different with the different dosage regimen when treated with the combination. Compared with the L-carnitine + 5-fluorouracil group, the apoptosis rate of cells was increased to (24.17 ± 3.12)% from (19.60 ± 1.06)% (P < 0.05). The G0/G1 phase proportion of cells was decreased to (62.62 ± 1.04)% from (72.95 ± 0.91)%, and the S phase proportion of cells was increased to (37.35 ± 1.03)% from (27.05 ± 0.91)% (P < 0.001). CONCLUSION: Treatment with L-carnitine and 5-fluorouracil could enhance the inhibitory effect of 5-fluorouracil on MGC803 cells. The possible mechanism of action is achieved by regulating the expression of Bcl-2 protein family and influencing the cell cycle.

Inhibitory effects on glucose metabolism of human gastric carcinoma MGC-803 cells induced by wogonin / 中国药科大学学报

@#To investigate the inhibitory effect of wogonin on glucose metabolism in human gastric carcinoma MGC-803 cells and its underlying mechanism, Amplex Red Glucose/Glucose Oxidase Assay Kit was used to explore the influence of wogonin on glucose uptake of MGC-803 cells and lactate assay kit was adopted to evaluate the lactate generation of MGC-803 cells. Annexin V/PI staining assay was performed to observe the apoptotic rate of MGC-803 cells. Changes in the expression of hexokinase 2(HK II), glucose transporter 1(GLUT1), pyruvate dehydrogenase kinase(PDHK)and lactate dehydrogenase A(LDH-A)were assessed by Western blot. These results demonstrated that wogonin significantly reduced glucose uptake of MGC-803 cells at certain concentration. Meanwhile, wogonin apparently reduced the lactate generation of MGC-803 cells, without inducing notable cell apoptosis. Besides, with the impact of wogonin, the expression of glucose metabolism related proteins declined obviously. In a word, wogonin could inhibit the glucose metabolism of MGC-803 cells without inducing apparent apoptosis, which may be related to its inhibitory influence on the proteins in glucose metabolism of MGC-803 cells.

Effects of a monomer purified from Paris Polyphylla (PP-22) on proliferation and apoptosis of human gastric carcinoma MGC803 cells / 中国药学杂志

OBJECTIVE: To investigate the effects of a monomer purified from Paris Polyphylla (PP-22) on proliferation, apoptosis of human gastric carcinoma MGC803 cells and to study the sensitizing effect of PP-22 on the proliferation of MGC803 cells to chemo-therapeutic drugs 5-fluorouracil (5-Fu) or lobaplatin. METHODS: MTT assay and cell colone formation inhibitory assay were used to determine the inhibitory effect of PP-22 on human gastric carcinoma MGC803 cells. The sensitizing effects of PP-22 on MGC803 cells to chemotherapeutic drugs 5-Fu, lobaplatin were determined by MTT assay. The percentage of apoptotic cells and cell cycle distribution were determined by flow cytometry. The expression of cell apoptosis associated proteins were analyzed by Western blotting. RESULTS: MTT assay showed that PP-22 inhibited gastric carcinoma MGC803 cells proliferation in dose-dependent manner (r=0.90, P<0.05), but did not inhibit the growth of normal liver L02 cells at the same concentration. Cell colony formation inhibitory assay demonstrated that cell clones decreased with the increase of drug concentrations. The chemosensitivity of 5-Fu, or lobaplatin combined with PP-22 was improved compared with PP-22 group, respectively. PI staining analysis showed that the cell cycle was arrested at S phase. Western blotting detecting showed that the expression of Caspase-9, Caspase-3 were downregulated, the anti-apoptotic protein Bcl-2 is downregulated and pro-apoptotic protein Bak upregulated. CONCLUSION: PP-22 inhibits proliferation and induces apoptosis of MGC803 cells effectively and also sensitizes MGC803 cells to chemotherapeutic drugs.

Anti-proliferation of Angong Niuhuang pill on tumor cells via inducement of apoptosis and down-regulation of mitochondrial membrane potential / 中国药理学与毒理学杂志

OBJECTIVE To validate the anticancer effect of Angong Niuhuang pill (AGNH) and pinpoint associated molecular mechanisms using human cancer cells.METHODS Human MGC-803 gastric carcinoma and human BEL-7402 hepatocarcinoma cells were incubated with AGNH 9,30,90,300 and 900 mg·L-1 for 24,48and 72 h,respectively.Cell viability was detected with 3-(4,5-dimethylthiazol-2-yl) -5-( 3-carboxymethoxyphe-nyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and colony formation assay.Apoptosis was measured with flow cytometry and Hoechst 33258/PI staining.Change in mitochondrial membrane potential (△qψ) was detected by spectrofluorophotometer.RESULTS AGNH inhibited MGC-803 cell proliferation ( for 48 h,r =0.996,P =0.002; for72 h,r=0.756,P=0.024 ) and BEL-7402 cells (for 48 h,r =0.732,P=0.030; for72 h,r=0.702,P =0.037) in a concentration-dependent manner,as showed by MTS assay.AGNH inhibited colony formation on MGC-803 cells (r =0.914,P =0.011 ) and BEL-7402 cells (r =0.871,P =0.024) in a concentration-dependent manner for 24 h.Hoechst 33258/PI staining and flow cytometry assay showed that AGNH 900 mg·L-1 for 24 h induced apoptosis of MGC-803 and BEL-7402 cells,and the apoptosis rate was 27.2％ and 19.7％,respectively.Compared with normal control group,AGNH 900 mg·L-1 for 3 min decreased the mitochondrial membrane potential of MGC-803 and BEL-7402 cells to 15.9％ and 15.0％ of control group.CONCLUSION AGNH inhibits proliferation of human cancer cells.Apoptosis and depolarization of mitochondrial transmembrane potential are probablly its mechanism.

Effect of activation of peroxisome proliferator-activated receptor γ on cell cycle distribution of human gastric carcinoma / 肿瘤

Objective: To explore the effect of activation of peroxisome proliferator-activated receptor gamma (PPARγ) on cell cycle arrest of gastric carcinoma cell line MGC803. Methods: The inhibitory effect of pioglitazone (PGZ) on proliferation of MGC803 cells was analyzed by MTT assay. Cell cycle were detected by flow cytometry (FCM). The protein expression of PPARγ, cyclinD1 and cell cycle protein-dependent kinase CDK4 in MGC803 cells was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Treatment with 0.1-10 μmol/L, PGZ for 96 h significantly inhibited cell proliferation. The proportion of MGC803 cells at G1 phase was significantly increased when treated with 10 μmol/L PGZ for 48, 72 and 96 h, and showed an apparent G1 phase arrest. The expression of PPARγ was at a low level in MGC803 cells and significantly up-regulated when treated with 10 μmol/L, PGZ for 48 h (P < 0.01). The expression of CDK4 in MGC803 cells was significantly down-regulated when treated with 10 μmol/L, PGZ for 96 h (P < 0.01) and the expression of cyclinD1 was slightly down-regulated. Conclusion: Activation of PPARγ significantly induces G1 phase arrest, which is associated with down-regulation of the expression of CDK4 and cyclinD1.

Up-expression of peroxisome proliferator-activated receptor-? and of retinoid X receptor-? enhances the inhibitory effect on tumor cells growth / 基础医学与临床

LOVO.PGZ significantly up-regulates the expression of PPAR? in MGC803 and LOVO cells,the expression of PPAR? was higher in combination group than PGZ alone(P

Study on Growth Effect and Apoptosis Action of Quercetin on Human Gastric Carcinoma Cells MGC-803 / 中国药房

OBJECTIVE:To study the effect of quercetin on the proliferation and apoptosis of human gastric carcinoma cell line MGC-803.METHODS:MTT assay was adopted to measure the effect of quercetin at different concentrations on the proliferation of MGC-803 cells.Cell apoptosis index(AI)induced by quercetin was measured by TUNEL assay.RESULTS: Quercetin at a certain concentration from 40?mol/L to 100?mol/L could successfully inhibit the proliferation of MGC-803 cells in a dose-dependent manner.TUNEL assay indicated that apoptosis cells induced by quercetin in treatment group were much more than that in control group after 48 hours(P

Celecoxib induces apoptosis in COX-2 non-expression gastric cancer cells / 中国病理生理杂志

AIM: To investigate whether celecoxib induces gastric cancer cell apoptosis in a COX-2 non-expression cell line.METHODS: The COX-2 protein was examined by western blotting.Fluorescence microscopy,DNA agarose gel electrophoresis and flow cytometry analysis were used to test apoptosis.RESULTS: COX-2 was expressed in AGS but not MGC-803 gastric cancer cell line;Selective COX-2 inhibitor celecoxib induced MGC-803 cell line apoptosis in a concentration and time-dependent manner.CONCLUSION: Celecoxib induces apoptosis in COX-2 non-expression gastric cancer cells.