ABSTRACT
Objective: To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.Method: A total of 40 SD rats were randomly divided into four groups:low-dose,middle-dose,high-dose MSJZT (0.213,0.426,0.853 g·kg-1) groups and normal group (n=10).The treatment groups were administrated through gastric perfusion,and the normal group was given the equivalent volume of normal saline for 10 days.1.5 h after the last treatment,chloral hydrate peritoneal anesthesia was performed,blood was collected from heart,and different doses of serum were separated to prepare drug-containing serum of low-dose,middle-dose,high-dose MSJZT groups,in order to incubate SGC-7901 gastric cancer cell.Early and late apoptosis rates were detected with flow cytometry.Afterwards,the tumor suppressor gene p53,c-nucleoprotein gene (c-Myc),cysteine-aspartic acid protease-3(Caspase-3),B-cell lymphoma-2(Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR).The protein expressions of p53,c-Myc,Caspase-3,Bcl-2 were detected by immunofluorescence.Result: Compared with the normal group,the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(PPPPPPConclusion: MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2,and promoting the expressions of pro-apoptotic-related molecules p53,c-Myc,Caspase-3.
ABSTRACT
@#Objective:To explore the mechanism of EYA1 (eyes absent 1) inhibiting the malignant progression of gastric cancer SGC7901 cells through regulating PTEN/PI3K/AKT signaling pathway. Methods: Twenty-nine pairs of gastric cancer tissues and para-cancerous tissues collected at the General Surgery center, Southwest Hospital Affiliated to Military Medical University during June 2016 and June 2018 were used in this study. Wb and RT-PCR assays were used to test the mRNA and protein expressions of EYA1 in gastric cancer tissues and the paired para-cancerous tissues; Transfection with plasmid or siRNAs were used to up-regulate or down-regulate EYA1 or PTEN expression in gastric cancer SGC-7901 cells; MTT, Flow Cytometry, Wound Healing and Transwell assays were carried out to detect cell proliferation, apoptosis, metastasis and invasion abilities, respectively. Results: EYA1 expression was decreased in gastric cancer tissues as compared with the para-cancerous tissues at both mRNA and protein levels (P<0.01); EYA1 over-expression significantly enhanced the proliferation, metastasis and invasion of SGC-7901 cells (all P<0.05), and inhibited cell apoptosis (P<0.05); moreover, its over-expressionsignificantly increased the expression of PTEN, and inhibited the activation of PI3K/AKT pathway (all P< 0.05 or P<0.01). However, the above effects mediated by EYA1 up-regulation were significantly impaired after the knockout of PTEN (all P<0.05 or P<0.01). Conclusion: EYA1 can inhibit the malignant progression of gastric cancer SGC-7901 cells through promoting the expression of PTEN and activating PI3K/AKT pathway.
ABSTRACT
Objective: To investigate the effects of silymarin on the proliferation, migration and invasion of human gastric cancer cell line SGC 7901 and its role in promoting apoptosis, and to clarify their possible mechanisms. Methods: The human gastric cancer SGC 7901 cells in logarithmic growth phase were selected and divided into control group (without silymarin) and different concentrations (15, 30, and 60 mg · L-1) of silymarin groups. Inverted microscope was used to observe the morphology of cells. MTT method was used to detect the cell cycle and the apoptotic rates of cells in various groups. The inhibitory rates of proliferation of cells in various groups were detected by flow cytametry. Scratch test and Trans well chamber assay were used to detect the cell migration and invasion abilities. Results: After the SGC 7901 cells were treated with silymarin for 24 h, compared with control group, the adherent densities of the cells in 30 and 60 mg · L-1 silymarin groups were smaller, the cell shape was irregular and the volume was smaller, resulting in a large number of cell debris. Compared with control group, the inhibitory rates of cells in different concentrations of silymarin groups were increased significantly (P<0. 05); the percentages of cells in Gi phase and the apoptosis rates of SGC 7901 cells were increased significantly (P<0. 05); the migration distances were decreased (P<0. 05) and the number of transmembrane cells was decreased significantly (P<0. 05). Conclusion: Silymarin can inhibit the proliferation of human gastric cancer SGC 7901 cells by inducing the G1 phase cell arrest and early apoptosis.
ABSTRACT
The purpose of this study was to investigate the inhibitory effect of the main 9,10-dihydrophenanthrene orchinol isolated from Spiranthes sinensis Radix et Herba on the invasion and migration of human gastric cancer SGC-7901 cells and its preliminary molecular mechanism. SGC-7901 cells were cultured in vitro, after the cells were treated with different final concentrations(5, 10, 20, 40, 80 μmol·L⁻¹) of orchinol for 24, 48 or 72 hours, the effect of orchinol on cell viability was measured by MTT assay. Wound healing and Transwell assays were performed to determine the effects of different final concentrations(5, 10, 20, 40 μmol·L⁻¹) of orchinol for 48 hour on invasion and migration abilities of SGC-7901 cells, respectively. The protein expression levels of β-catenin, Wnt-3α, DvL2, cyclinD1 and GSK-3β were detected by Western blot. The results showed that 5-80 μmol·L⁻¹ orchinol inhibited the viability of SGC-7901 cells in a dose-dependent and time-dependent manner, and the IC₅₈ values of 24, 48 and 72 hours were 77.79, 42.96 and 7.85 μmol·L⁻¹, respectively. Compared with the control group, the ability of invasion and migration of SGC-7901 cells was significantly inhibited after treated with 5, 10 and 20 μmol·L⁻¹ orchinol for 48 hours (<0.05, <0.01), and the dose-effect relationship was observed. The results of Western blot showed that orchinol could significantly down-regulate the protein expression levels of β-catenin, Wnt3a, DvL2 and cyclinD1, and up-regulate the protein expression level of GSK-3β(<0.05, <0.01, <0.001). The above results suggest that orchinol can obviously inhibit the invasion and migration of SGC-7901 cells, which may be related to its inhibition of Wnt3a/β-catenin signaling pathway and the proteins expression of downstream genes.
Subject(s)
Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Phenanthrenes , Stomach Neoplasms , Wnt Signaling Pathway , Wnt3A Protein , beta CateninABSTRACT
AIM: To explore the effect of microRNA-146a (miR-146a) on apoptosis of human gastric cancer SGC-7901 cells and the underluing mechanism.METHODS: miR-146a mimic (up-regulated miR-146a expression) and miR-146a inhibitor (down-regulated miR-146a expression) were transfected into the SGC-7901 cells by liposome method.At the same time, miRNA nonsense sequence transfection group as the negative control group (NC group) was set up.RT-qPCR was used to evaluate the levels of miR-146a in the SGC-7901 cells after transfection.The effects of miR-146a on the cell apoptosis and growth were assessed by flow cytometry analysis and CCK-8 assay, respectively.The effect of over-expression or knockdown of miR-146a on transforming growth factor-β-activated kinase 1 (TAK1)/ nuclear factor-kappa B (NF-κB) signaling was evaluated by RT-qPCR and Western blot.RESULTS: miR-146a modulated apoptosis of SGC-7901 cells.Over-expression of miR-146a significantly increased apoptosis, whereas knockdown of miR-146a inhibited the apoptosis of SGC-7901 cells.The expression of TAK1 at mRNA and protein levels was significantly decreased when miR-146a mimic was transfected into the SGC-7901 cells (P<0.05).On the contrast, the expression of TAK1 at mRNA and protein were significantly higher in miR-146a inhibitor transfection group than that in NC group (P<0.05), suggesting that miR-146a negatively regulated TAK1 expression.Moreover, knockdown of TAK1 enhanced the apoptosis of SGC-7901 cells (P<0.01), while over-expression of TAK1 inhibited the apoptosis of SGC-7901 cells(P<0.01).Additionally, both over-expression of miR-146a and knockdown of TAK1 led to a prominent increase in the expression of NF-κB inhibitor protein alpha (IκBα) and a significat decrease in B cell lymphoma-2 (Bcl-2) level in the SGC-7901 cells.CONCLUSION: miR-146a significantly promotes apoptosis of SGC-7901 cells by inhibition of NF-κB pathway via targeting TAK1.
ABSTRACT
OBJECTIVE: To investigate the effect and mechanism of juglone on the apoptosis of human gastric cancer SGC-7901 cells. METHODS: The antiproliferative effect of juglone on SGC-7901 cells was tested by the MTT assay. The apoptosis rate and in-tracellular reactive oxygen species(ROS) level were detected by flow cytometry (FCM). The expression of JNK, p-JNK, p38, and p-p38 proteins were examined by Western blot. In order to clarify the role of ROS in the apoptosis induced by juglone on SGC-7901 cells, the combination of the juglone and ROS inhibitor NAC groups were set up in each experiment above. RESULTS Juglone could effectively inhibit the proliferation of SGC-7901 cells (IC50 for 72 h was 24.16 μmol·L-1). When combination juglone with NAC, the IC50 raised to 36.91 μmol·L-1. After 72 h of exposure to 5-20 μmol·L-1 of juglone, the cell apoptosis rate increased gradually with the increase of juglone concentration. After adding NAC, the apoptosis rates declined and the apoptosis rate of 20 μmol·L-1 group decreased from 32.06% to 11.56%. After SGC-7901 cells were treated with juglone for 24 h, the ROS level increased and mitochondrial transmembrane potential decreased which were inhibited by the pretreatment of NAC. After 48 h of exposure to different con-centration of juglone, the expressions of p-p38 and p-JNK proteins were up-regulated which could also be inhibited by the adding of NAC. Meanwhile, there were no significant changes in p38 and JNK protein expression in all groups. CONCLUSION: Juglone can induce apoptosis of human gastric cancer SGC-7901 cells by JNK and P38 pathway mediated by reactive oxygen species.
ABSTRACT
Objective To study effect of a novel schiff base ruthenium coordination compound on cell proliferation and apoptosis of gastric cancer SGC-7901 cell.Method Gastric cancer SGC-7901 cell were divided into four groups according to different treatment of novel schiff base ruthenium coordination compound (concentration of 10, 30, 50μmol/L) and blank group with DMSO.Cell proliferation was detected by MTT assay, cell apoptosis and cell cycle were analysed by flow cytometry.ResuIts MTT results showed the inhibitory effect of novel schiff base ruthenium coordination compound on SGC-7901 cell enhanced with the increase of its concentration, and inhibitory rates were higher than that of blank group at 24, 48, 72 h.Flow cytometry results showed the apoptosis rate of novel ruthenium coordination compound groups of 10, 30, 50μmol/L were (17.64 ±1.21)%, (26.47 ± 0.61)%, (55.63 ±1.49)%, respectively, all higher than that of blank group (P<0.05).The cell proportion of G1 phase increased with the increasing of the novel schiff base ruthenium coordination.ConcIusion A novel schiff base ruthenium coordination compound could inhibit the growth of gastric cancer SGC-7901 cells, promote apoptosis and arrest gastric cancer SGC-7901 cells at G1.
ABSTRACT
Objective To observe the effect of Xinjia-Liangfu(XJLF)formula on human gastric cancer(SGC-7901)tumor growth in nude mice and its impact on survivinand Caspase-3 expression in tumor tissue. Methods The animal model of SGC-7901 xenografted nude mice was established, and all the mice were randomly divided into 6 groups, namely model control group, 5-Fu group, XJLF formula high-dose group, XJLF formula medium-dose group , XJLF formula low-dose group and drug combination group(XJLFformula medium-dose and 5-Fu). After 10 days of continuous treatment, tumor inhibition rate was calculated and the expression of Survivin and Caspase-3 was detected by immunohistochemistry techniques. Results The tumor inhibition rates of 5-Fu group, XJLF formula high-dose group, XJLF formula medium-dose group, XJLF formula low-dose group and drug combination group were 55.03%, 56.38%, 41.23%, 23.09%and 60.28%respectively, and the tumor inhibition rate of drug combination group was significantly increased compared to the model control group(P<0.05), 5-Fu group(P<0.05)and XJLF high-dose group(P<0.05) Meanwhile, the Caspase-3 expression was significantly up-regulated and the Survivin expression was significantly down-regulated in drug combination group(Caspase-3 was 77 539.74±50 912.5, Suvivin was 35 709.1±10404.4)compared to the model control group(Caspase-3 was 18 464.71±7 537.01,Suvivin was 63 449.4±50 498.8), P<0.05 or 0.01, 5-Fu groupCaspase-3 was (47 198.89±10 751.2), Suvivin was(37 016.8±6 024.4), P<0.05 or 0.01and XJLF formula high-dose groupCaspase-3was (44213.57±7041.6), Suvivin was (38811.1±7313.0)respectively. Conclusion The tumor inhibition rate of drug combination group was higher than XJLF formula high-dose group and 5-Fu group. Also combination group hadstronger effect compared to model control group in terms of down-regulation of Caspase-3 expression and up-regulation of Survivin expression, which indicates a benefit ofXJLF and 5-Fu combination treatment.
ABSTRACT
Objective To study the effects and mechanism of Octreotide to inhibit the proliferation of human gastric cancer cells in vitro . Methods Human gastric cancer cell line SGC 7901 was treated with Octreotide. Human fibroblast cell line HF and 5 FU were used as control. MTT assay and fluorescent microscopy as well as flow cytometry were performed in this study. Results Octreotide inhibited the growth of SGC 7901 in vitro within certain concentrations. The suppression was quantity dependent but did not occur when up to a certain concentration. There was no difference between Octreotide and 5 FU in their inhibition on SGC 7901. Octreotide had no effects on normal human fibroblast cell line HF. When SGC 7901 was treated with Octreotide, the typical apoptotic bodies were identified by flow cytometry and fluorescent microscopy. Conclusion Octreotide can inhibit the proliferation of human gastric cancer cell line SGC 7901 in vitro . The induction of apoptosis by Octreotide might be the primary mechanism.
ABSTRACT
Objective To explore the effects of humulon on kinetic parameters of N-acetyltransferase-1(NAT1) of human gastric cancer SGC-7901.Methods Employing HPLC,using para-aminobenzoic acid(PABA) as substrate,in intact SGC-7901 cells and their cytoplasm,taking the speed of PABA being acetylated to Ac-PABA by NAT1 as the rate of NAT1,using double reciprocal plot,taking the reciprocal of concentration of PABA and reaction rate of NAT1 as coordinates,regression equation was obtainied and the Michaelis constant(Km) and maximum reaction velocity(Vmax) were calculated.Results Study on enzyme kinetics demonstrated,as for intact SGC-7901 cells,Km and Vmax of control group were(3.910?0.087) ?mol/L and(0.306 0?0.006 7) pmol/L(1?106 cells),respectively,Km and Vmax of the humulon group were(3.830?0.123) ?mol/L and(0.275 0?0.005 8) pmol(1?106 cells),respectively.As for the cytoplasm of SGC-7901 cells,Km and Vmax of control group were(760.2?210.2) ?mol/L and(0.191 0?0.043 7) pmol/(mg?min),Km and Vmax of the humulon group were(449.0?72.9) ?mol/L and(0.094 0?0.010 4) pmol/(mg?min).Statistically,as for intact SGC-7901 cells or their cytoplasm,there was no difference of the Km between control group and humulon group,but there was remarkable difference of Vmax between control group and humulon group,P
ABSTRACT
Objective To investigate the relationship between the antiproliferation effects of aloe-emodin on growth of gastric cancer cells and cell cycle arrest.Methods Human gastric cancer SGC-7901 cells were treated with 2.5,5,10,20,and 40 ?mol/L aloe-emodin for 1—5 d.The cell growth was determined by MTT assay.Cell proliferation and cycle distributions were analyzed by flow cytometry.Western blotting assay was used to detect the changes of cell cycle regulators,cyclins,and cyclin-dependent kinases(CDK).Results Aloe-emodin inhibited the growth of gastric cancer cells in a dose-dependent manner.Treatment of aloe-emodin resulted in cell cycle arresting at G2/M phase.Its molecular mechanisms involved the decrease of the expression of cyclin A and CDK2,the increase of the expression of cyclin B1 and CDK1.Conclusion One of the antitumor mechanism of aloe-emodin on the growth of gastric cancer SGC-7901 cells is to arrest the cell cycle,which indicates that aloe-emodin has a potential value for the treatment of gastric cancer in clinic.
ABSTRACT
AIM: To study the effect of Zhongjiefeng Injection(Herba sarcandrae) on mouse liver cancer H_22,and its activities on gastric cancer SGC-7901 in vitro,to find out its influence on the cell cycle. METHODS: By injecting H_22 tumor cells in vivo into mice to check its inhabition on entity and ascites tumor.MTT colorimetric method was used to study the anti-tumor activity on gastric cancer SGC-7901,then we calculated its IC_50 and applied the flow cytometry(FCM) to detect its influence of cell cycle. RESULTS: The rates of inhabition on H_22 entity tumor with three concentrations were 29.8%,36.2%,40.5%,and was the remarkable different from the model group(P