Effects of gypenosides on proliferation and apoptosis of human gastric cancer cells and its mechanism / 中国药理学通报

Aim To explore the effect of gypenosides on proliferation and apoptosis of human gastric cancer cells SGC-7901 and AGS and its mechanism. Methods Different concentrations of gypenosides were cultured with human gastric cancer cells SGC-7901 and AGS. Cell viability assay was used to detect cell proliferation activity, and the IC

Potential mechanism of Atractylodes macrocephala aqueous extract inhibiting gastric carcinoma through PI3K-Akt-NF-κB signaling pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨白术（Atractylodes macrocephala）水提物抑制胃癌SGC-7901细胞活性的潜在机制。方法：分别使用蒸馏水（对照）和白术水提物（白术治疗组）灌胃SD大鼠后，采集静脉血后分离其血清、过滤并分别命名为对照组血清（CON-S）和白术组血清（AM-S）。将胃癌SGC7901细胞分为对照组、10% AM-S组和20% AM-S组，其中两个AM-S组细胞分别在相应浓度的AM-S血清中培养24 h，对照组细胞用正常培养基培养相同时间，收取SGC7901细胞和上清液用于进一步分析。使用MTT法检测各组细胞活力，通过商业试剂盒测定乳酸脱氢酶（LDH）、丙二醛（MDA）和超氧化物歧化酶（SOD）的水平，采用ELISA试剂盒检测各组细胞中IL-6和TNF-α的含量，采用WB法评估各组细胞中PI3K-Akt-NF-κB信号通路相关蛋白的表达。结果：10% AM-S组和20% AM-S组的SGC7901胃癌细胞增殖活力相较于对照组分别降低48.9%和53.25%（P<0.05或P<0.01）；胃癌细胞上清液中，相较于对照组，10% AM-S组和20% AM-S组LDH水平分别升高29.25%和123%、SOD活性分别升高18%和54.60%、MDA水平分别降低27.8%和40.0%，IL-6水平分别降低15%和17.5%、TNF-α水平分别降低29.71%和40.16%（P<0.05或P<0.01）。相较于对照组，AM-S组中PI3K-Akt-NF-κB信号相关蛋白的水平显著下降（P<0.05或P<0.01）。结论：白术水提物可以通过抑制癌细胞增殖活力、促进凋亡、抑制肿瘤微环境中的促炎因子分泌以及改变细胞内的氧化应激水平等方式抑制胃癌，其机制可能是通过抑制PI3K-Akt-NF-κB通路来实现这些抗癌作用的。

Expression of β-1,6 glucosaminyl (N-acetyl) transferase 2 in gastric cancer and its clinical significance / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨β-1,6 N-乙酰氨基葡萄糖转移酶2（GCNT2）基因在胃癌（GC）组织中的表达及其在GC发生、发展和诊断及预后中的作用。方法：利用TIMER、GEPIA2、Oncomine和UALCAN等数据库数据，以及2018年1月至2019年12月滨州医学院附属医院手术切除的25例GC患者的癌和配对癌旁组织标本，分析GCNT2基因在GC组织中的表达及其在GC诊断和预后中的价值，利用LinkedOmics、GSEA和ssGSEA分析GCNT2所涉及的主要信号通路及其与免疫浸润之间的相关性。将pc-GCNT2及其阴性对照质粒转染进胃癌SGC-7901和BGC-823细胞，用克隆形成实验和Transwell实验检测GCNT2对GC细胞增殖和侵袭的影响，WB法检测细胞中GCNT2、STAT3和PD-L1蛋白的表达水平。结果：GCNT2 mRNA在GC组织中的表达水平显著低于癌旁组织（P<0.05或P<0.01），其表达水平与患者预后显著相关（P<0.05），其对GC诊断有较高的价值。GCNT2在GC组织中的甲基化状态显著高于癌旁组织，GCNT2基因参与的生物过程主要是参与细胞形态发生的成分、细胞间黏附、多细胞生物信号和突触传递等。单基因GSEA分析发现，GCNT2在GC中主要抑制IL-6/JAK/STAT3、IL-2/STAT5信号通路和炎症反应、α/γ干扰素响应与NF-κB表达等。GCNT2的表达与GC组织的免疫浸润具有显著相关性。过表达GCNT2可显著抑制GC细胞的增殖和侵袭能力（均P<0.01），降低细胞中STAT3和PD-L1的表达水平（均P<0.01）。结论：GCNT2基因在GC组织中低表达，与GC的诊断及预后显著相关，其主要通过抑制IL-6/JAK/STAT3和免疫相关致癌信号通路而在GC的发生、发展中发挥重要的作用。

Clinical significance of LINC00997 expression in gastric cardia adenocarcinoma and its effect on migration, invasion and epithelial-mesenchymal transition of SGC7901 cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：检测LINC00997在胃贲门腺癌（GCA）组织及胃癌细胞中的表达，分析其表达与患者临床病理特征和预后的关系，探讨敲减LINC00997对胃癌SGC7901细胞迁移、侵袭及上皮间质转化（EMT）的影响。方法：基于TCGA和GTEx数据库分析LINC00997在胃癌组织中的表达及其与患者预后的关系。应用qPCR法检测68例GCA组织和相应癌旁组织以及胃癌细胞中LINC00997的表达水平，分析其表达与患者临床病理特征及预后的关系。通过划痕愈合、Transwell侵袭实验分别检测敲减LINC00997对SGC7901细胞迁移和侵袭的影响，qPCR法和WB法检测敲减LINC00997对EMT相关标志物E-cadherin、N-cadherin及vimentin表达的影响。结果：LINC00997在胃癌组织中的表达水平显著高于癌旁组织（P<0.05），且LINC00997高表达组患者的OS及DFS显著低于LINC00997低表达组患者（P<0.01或P<0.05）。在68例在GCA组织中，LINC00997的表达水平显著高于癌旁组织（P<0.01），其表达与患者淋巴结转移、TNM分期及OS相关联（P<0.05或P<0.01）。敲减LINC00997的SGC7901细胞的迁移和侵袭能力均显著降低（均P<0.01），细胞中E-cadherin的表达显著升高，N-cadherin、vimentin的表达均显著降低（P<0.05或P<0.01）。结论：LINC00997在GCA组织和胃癌细胞中高表达，其高表达可能促进了胃癌细胞的迁移、侵袭及EMT进程，有望成为GCA患者预后评估的分子标志物。

Effect of Circular RNA hsa_circ_0067582 on the Proliferation and Invasion Ability of Gastric Cancer Cells / 中国医学科学院学报

Objective To investigate the effects on cell proliferation and invasion of the circular RNA hsa_circ_0067582 in gastric cancer(GC). Methods After hsa_circ_0067582 overexpression (Oe-circ_0067582) plasmid was transfected into AGS and SGC-7901 cells,the cell viability,proliferation,invasion ability,and apoptosis were detected by CCK-8,colony formation and EdU assays,Transwell assay,and flow cytometry,respectively.Western blotting was employed to detect the expression levels of proteins related to the cell apoptosis and epithelial-mesenchymal transition(EMT).The effect of Oe-circ_0067582 on the growth of SGC-7901 cells in nude mice was observed.Bioinformatics tools were used to predict the binding target miRNA of hsa_circ_0067582,and the competing endogenous RNA(ceRNA)regulatory network was established.Finally,functional enrichment was performed to analyze the biological functions of the target genes of the predicted miRNA. Results Compared with the pLO-ciR(empty plasmid)group,the Oe-circ_0067582 group in AGS and SGC-7901 cells attenuated the cell viability(t=7.883,P=0.001;t=5.679,P=0.005),proliferation(t=6.709,P=0.003;t=5.857,P=0.003),and invasion ability(t=7.782,P=0.002;t=6.342,P=0.003)and induced cell apoptosis(t=7.225,P=0.002;t=11.509,P=0.001).Western blotting showed that the Oe-circ_0067582 group in AGS and SGC-7901 cells up-regulated the protein levels of cysteinyl aspartate specific proteinase (Caspase) 3(t=6.863,P=0.002;t=7.024,P=0.001),Caspase 7(t=3.295,P=0.04;t=6.008,P=0.004),Caspase 9(t=4.408,P=0.012;t=6.278,P=0.004),and E-cadherin(t=12.453,P=0.002;t=10.867,P=0.001),while down-regulated those of Vimentin(t=7.242,P=0.002;t=5.694,P=0.004)and N-cadherin(t=6.480,P=0.003;t=7.446,P=0.001).Furthermore,Oe-circ_0067582 significantly inhibited the growth of tumor in the SGC-7901 tumor-bearing nude mice(t=3.526,P=0.017).The prediction based on TargetScan and miRnada suggested that hsa_circ_0067582 can competitively bind to hsa-miR-181b-3p,hsa-miR-337-3p,hsa-miR-421,and hsa-miR-548d-3p.The functional enrichment indicated that the target genes of miRNA were involved in multiple cancer-related biological processes including negative regulation of apoptotic process,gene expression,transcriptional misregulation in cancer,transforming growth factor-β,and p53 signaling pathways. Conclusion Oe-circ_0067582 can inhibit the proliferation and attenuate EMT process to reduce the invasion ability of AGS and SGC-7901 cells,which provides a new target for the treatment of GC.

miR-153-3p regulates proliferation, invasion and migration of gastric cancer SGC7901 cells via targeting FZD3 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨miR-153-3p对胃癌SGC7901细胞增殖、侵袭和迁移的作用及其机制。方法：收集2018年5月至2020年6月宁夏医科大学总医院收治的60例胃癌患者的癌和配对癌旁组织标本，以及人胃癌细胞系NCI-N87、AGS、SNU-5、SGC7901和胃上皮细胞GES-1，qPCR法检测miR-153-3p在胃癌组织与细胞中的表达水平。将miR-153-3p mimic及mimic对照序列转染至SGC7901细胞，用CCK-8、克隆形成、流式细胞术、TUNEL、Transwell和划痕愈合实验分别检测上调miR-153-3p对SGC7901细胞增殖、凋亡、侵袭和迁移的影响。构建裸鼠SGC7901细胞移植瘤模型，观察miR-153-3p对肿瘤生长的影响。通过生物信息学数据库和双荧光素酶报告基因实验预测并验证miR-153-3p与FZD3靶向关系，WB法检测miR-153-3p对FZD3蛋白及Wnt/β-catenin通路相关蛋白表达的影响。结果：miR-153-3p在胃癌组织和细胞中表达水平分别显著低于癌旁组织和GES-1细胞（均P<0.01），以SGC7901细胞中表达水平最低。上调miR-153-3p显著抑制SGC7901细胞的增殖、侵袭和迁移能力，并提高细胞凋亡率（均P<0.01），同时上调细胞中E-cadherin表达而下调N-cadherin、MMP2和MMP9表达（均P<0.01）。在体内实验表明，静脉注射miR-153-3p mimic显著降低移植瘤体积和瘤组织中Ki-67表达而上调P57表达（均P<0.01）。机制分析表明，miR-153-3p靶向结合FZD3基因的3′UTR区域，上调miR-153-3p会抑制FZD3表达并上调β-catenin、TCF-4和cyclin D1水平（均P<0.01）。结论：miR-153-3p靶向FZD3并通过Wnt/β-catenin信号通路调控胃癌SGC7901细胞的增殖、侵袭和迁移。

Screening and preliminary analysis of differentially expressed miRNAs in gastric cancer cell-derived exosomes induced by Helicobacter pylori / 中华微生物学和免疫学杂志

Objective:To investigate the differentially expressed microRNAs (miRNAs) in human gastric carcinoma SGC-7901 cell-derived exosomes induced by Helicobacter pylori ( H. pylori), providing new clues for further elucidating the carcinogenic mechanism of H. pylori. Methods:Ultracentrifugation and exosome extraction kit were used to extract the exosomes released by the H. pylori-stimulated and negative control group, and transmission electron microscope(TEM), nanoparticle tracking analysis(NTA) and Western blot experiments were employed to identify exosomes. Then, exosomes were labeled with the fluorescent dye PKH67 and co-cultured with THP-1-derived macrophages. The internalization of exosomes by macrophages was observed by laser confocal fluorescent microscopy. Additionally, miRNA microarray chips were performed to detect the differentially expressed miRNAs of exosomes from the two groups of cells. Real-time fluorescence quantitative PCR (qRT-PCR) was used to verify the expression of four differentially expressed miRNAs. Furthermore, the target genes and their functions as well as the possible signal pathways involved of partial differentially expressed miRNAs were predicted and analyzed by bioinformatics software. Differentially expressed miR-382-5p was labeled by Cy3 to observe whether it could be transferred to macrophages through exosomes. The expression of phenotype molecule CD206 and the cytokines TNF-α, IL-6 and IL-10 in miR-382-5p mimic-transfected macrophages were analyzed by qRT-PCR and ELISA, and the proportion of cells expressing CD206 and HLA-DR was analyzed by flow cytometry. Results:The extracted exosomes were consistent with exosome morphology and highly expressed the surface marker proteins CD9, CD63 and TSG101. After co-culturing with THP-1 derived macrophages for 12 h, the exosomes could be internalized by macrophages. Compared with the control group, there were 130 up-regulated miRNAs and 111 down-regulated miRNAs in the H. pylori-stimulated group. Bioinformatic analysis showed that the potential target genes of partial differentially expressed miRNAs were mainly involved in the regulation of PI3K-AKT, NF-κB, JAK-STAT, stem cell pluripotency and other inflammation and tumor-related pathways. miR-382-5p could be transferred to macrophages through exosomes, and induced the expression of M2-type phenotype molecule CD206 and cytokines IL-10 in macrophages, while inhibited the expression of TNF-α and IL-6 and increased the proportion of CD206 high HLA-DR low cells. Conclusions:H. pylori treatment caused a significant change in the expression level of exosome miRNAs in SGC-7901 cells. Bioinformatics analysis demonstrated that the prospective targets of these differentially expressed miRNAs might play an important role in the regulation of inflammation and tumor-related signaling pathways. miR-382-5p might induce the M2-type polarization of macrophages.

miR-361-5p reverses oxaliplatin resistance of gastric cancer SGC-7901 cells by targeting CCND1 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects of miR-361-5p on the oxaliplatin (OXA) resistance of gastric cancer SGC-7901 cells and its mechanism. Methods: The expression of miR-361-5p in gastric cancer cells (MKN-45, MGC80-3 and SGC-7901) and drug-resistant SGC-7901/OXA cells was detected by qPCR. The SGC-7901/OXA cells were transfected with miR-361-5p mimics/inhibitor or sh-CCND1 by using Liposome transfection technology. Then, cell proliferation, apoptosis and cell cycle of SGC-7901/OXA cells were measured by CCK-8 assay and Flow cytometry, respectively. The targeting relationship between miR-361-5p and CCND1 was examined by Dual luciferase report gene assay. The expression level of CCND1 in SGC-7901/OXA cells was detected by WB. Results: miR-361-5p was down-regulated in multiple gastric cancer cells and SGC-7901/OXA cells (P<0.05 or P<0.01). Over-expression of miR-361-5p significantly promoted the apoptosis, induced G0/G1 cell cycle arrest and inhibited cell proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Dual luciferase reporter gene results verified that miR-361-5p targeted CCND1 and negatively regulated its expression (P<0.01). Further experiments showed that targeted down-regulation of CCND1 induced apoptosis and G0/G1 cell cycle arrest and inhibited CCND1 expression and proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Over-expression of miR-361-5p targetedly down-regulated CCND1 and further promoted cell apoptosis, induced G0/G1 cell cycle arrest and inhibited cell proliferation of SGC-7901/OXA cells (P<0.05 or P<0.01). Conclusion: miR-361-5p over-expression can reverse the resistance of SGC-7901/OXA cells to OXA, and the mechanism may be related to its targeted down-regulation of CCND1 expression.

Expression of forkhead box protein D3 in gastric cardia adenocarcinoma tissues and its effect on biological behaviors of SGC-7901 cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨叉头框蛋白D3（forkhead box protein D3，FOXD3）在胃贲门腺癌（gastric cardia adenocarcinoma，GCA）中的表达及其对SGC-7901细胞生物学行为的影响。方法：从河北医科大学第四医院生物标本库中选取2014年6月至2016年12月手术切除的49例GCA组织及相应癌旁组织标本，qRT-PCR检测FOXD3在GCA组织、癌旁组织以及在5种胃癌细胞系中（BGC-823、SGC-7901、HGC-27、MGC-803及NCI-N87）的表达。向SGC-7901细胞转染pc-DNA3.1-FOXD3或pc-DNA3.1，采用细胞增殖实验、克隆形成实验、划痕愈合实验和Transwell小室侵袭实验分别检测FOXD3过表达对SGC-7901细胞增殖、克隆形成、迁移和侵袭的影响，qRT-PCR及WB法检测细胞转染前后上皮-间质转化（epithelial-mesenchymal transition，EMT）相关分子mRNA及蛋白的表达情况，流式细胞术检测转染前后细胞周期改变。结果：GCA组织中FOXD3 mRNA的表达量明显降低，其表达水平与患者临床分期和淋巴结转移密切关联；FOXD3在胃癌细胞系中的表达均低于正常细胞（均P<0.01）。FOXD3过表达能明显抑制SGC-7901细胞的增殖、克隆形成、迁移和侵袭能力（均P<0.01），提高SGC-7901细胞中E-cadherin的表达水平，减少N-cadherin、β-catenin和vimentin的表达水平（均P<0.01），使细胞周期阻滞在G0/G1期（P<0.01）。结论: FOXD3在GCA组织中的表达明显下调，其过表达可以抑制胃癌细胞的生物学行为，FOXD3可作为抑癌基因为肿瘤治疗提供新思路。

Effects of usnicoyinamide on proliferation, apoptosis, cell cycle, and migration of SGC-7901 gastric cancer cells / 中草药

Objective: To investigate the inhibitory effect of usnicoyinamide on the proliferation of gastric cancer cell line SGC-7901 and its mechanism. Methods: SGC-7901 cells were cultured in vitro and divided into two groups: control group and experimental group with different concentrations of usnicoyinamide. The morphology of each group of cells was observed by a microscope; Proliferation of SGC-7901 cells was measured by MTT assay; The mechanism of apoptosis was studied by AnnexinV/PI double staining and DAPI fluorescence staining; Flow cytometry was used to detect the effect of usnicoyinamide on the cell cycle; Effect of usnicoyinamide on invasion and migration of SGC-7901 cells was detected by cell scratch test. Results: After SGC-7901 cells were treated with usnicoyinamide, the cells were wrinkled, deformed and adherent cells fell off; The results of MTT showed that the inhibition of the proliferation of SGC-7901 cells was a significant dose-effect relationship and time-dependent; The results of AnnexinV/PI double staining showed that nicotine increased the late apoptosis rate of SGC-7901 cells, and DAPI staining showed obvious nuclear concentration and nuclear fragmentation of apoptosis. The results of flow cytometry showed that the cell cycle of SGC-7901 cells stagnated in S phase; Scratch test showed that the mobility of SGC-7901 cells was decreased more obviously with the prolongation of time and the increase of concentration. Conclusion: Usnicoyinamide can inhibit the proliferation of gastric cancer cell line SGC-7901, and its mechanism may be achieved by inducing late apoptosis, inducing S phase cell arrest and inhibiting the invasion and migration of SGC-7901 cells.

Gambogic acid suppresses invasion in human gastric carcinoma SGC-7901 cells by modulating inhibitor of nuclear factor kappa-B kinase α/p65 signaling pathway in vitro / 解剖学报

Objective To investigate the effect of gambogic acid (GA) on invasion in human gastric carcinoma SGC-7901 cells and its possible mechanism. Methods Cell counting kit-8(CCK-8) assay was performed to detect the effects of GA, inhibitor of nuclear factor kappa-B kinase( IKK) 16 and 5-fluorouracil (5-FU) on cell activity of GES-1 and SGC-7901 cells. Cell invasion was assessed with Transwell invasion assay. Western blotting was used to analyze the protein levels of vimentin, matrix metalloproteinase 2 ( MMP-2) and MMP-9 and protein phosphorylation of IKKα and p65. Results The cell activity was significantly decreased in SGC-7901 cells treated with GA in a dose-dependent manner with a half inhibiton concentration(IC50) value of 1. 89 μmol/L. But GA had no significant influence on cell viability of GES-1 cells. Meanwhile, 5-FU reduced the cell activity of GES-1 and SGC-7901 cells with IC50values of 7.36 μmol/L and 199.57 μmol/L respectively. Low-dose GA and IKK 16 impaired separately the ability of invasion in SGC-7901 cells, and down-regulated the protein levels of MMP-2, MMP-9 and vimentin, and inhibited phosphorylation of IKKot and p65, while a stronger inhibition was showed when the combination of GA and IKK16 was used. Conclusion Low-dose GA might inhibit invasion of SGC-7901 cells via IKKot/p65 signaling pathway.

Apatinib enhances the anti-tumor effect of cisplatin on gastric cancer by inhibiting HMGA2 / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of apatinib (APA) combined with cisplatin (DDP) on the proliferation, invasion and migration capacity of gastric carcinoma (GC) cells and its molecular mechanism. Methods: Cancer and para-cancerous tissue samples resected from 50 GC patients, who were surgically treated in Wuwei People's Hospital from January 2016 to June 2019, were collected for this study; in addition, GC cell lines MGC803 and SGC7901 were also collected. qPCR was used to detect the HMGA2 expression in tissues and mRNA expressions of molecules related to cell proliferation, migration and invasion in GC cell lines. MGC803 and SGC7901 cells were transfected with pcHMGA2 by liposome transfection technology. After treatment with DDP and APA at different concentrations, the cells were divided into NC, pcHMGA2, pcHMGA2+DDP and pcHMGA2+DDP+APA groups. Protein expression of HMGA2 in GC cells was detected by Western blotting, and proliferation, migration and invasion of the cells were detected by MTT and Transwell assay, respectively. Results: The mRNA expression of HMGA2 in GC tissues was higher than that in para-cancerous tissues (P<0.05), and the survival rate of GC patients in the high expression group was significantly reduced (P<0.01). DDP significantly inhibited the proliferation, invasion and migration of MGC803 and SGC7901 cells (all P<0.01); the proliferation, invasion and migration of MGC803 and SGC7901 cells in DDP+APA group significantly decreased (all P<0.01) as compared with DDP group; APA significantly enhanced the inhibitory effect of DDP on HMGA2 expression in GC cells (P<0.01); APA enhanced the anticancer activity of DDP against GC by down-regulating HMGA2 expression. Conclusion: APA promotes the anticancer activity of DDP against GC, and its molecular mechanism is the promotion of the inhibitory effect of DDP on HMGA2 expression.

Crebanine N-oxide, a natural aporphine alkaloid isolated from Stephania hainanensis, induces apoptosis and autophagy in human gastric cancer SGC-7901 cells

Objective: To investigate the cytotoxic effects and the potential mechanisms of crebanine N-oxide in SGC-7901 gastric adenocarcinoma cells. Methods: The cytotoxicity of crebanine N-oxide was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and cellular morphology was observed under a microscope. Cell apoptosis was determined by flow cytometry using propidium iodide staining. The expression levels of apoptotic-related proteins, cleaved caspase-3, cytochrome C, p53 and Bax, and autophagyrelated proteins p62, beclin1 and LC3 were detected by Western blotting assays. Results: Crebanine N-oxide treatment significantly inhibited the proliferation of SGC-7901 cells in a dose-dependent and timedependent manner via induction of G2-phase cell cycle arrest, apoptosis, and autophagy in SGC-7901 cells. Conclusions: Crebanine N-oxide could inhibit the growth of gastric cancer cells by promoting apoptosis and autophagy and could be used as a potential agent for treating gastric cancer.

Crebanine N-oxide, a natural aporphine alkaloid isolated from Stephania hainanensis, induces apoptosis and autophagy in human gastric cancer SGC-7901 cells

Objective: To investigate the cytotoxic effects and the potential mechanisms of crebanine N-oxide in SGC-7901 gastric adenocarcinoma cells. Methods: The cytotoxicity of crebanine N-oxide was evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and cellular morphology was observed under a microscope. Cell apoptosis was determined by flow cytometry using propidium iodide staining. The expression levels of apoptotic-related proteins, cleaved caspase-3, cytochrome C, p53 and Bax, and autophagy-related proteins p62, beclin1 and LC3 were detected by Western blotting assays. Results: Crebanine N-oxide treatment significantly inhibited the proliferation of SGC-7901 cells in a dose-dependent and time-dependent manner via induction of G

EYA1 inhibits malignant biological behavior of gastric cancer SGC-7901 cells by regulating PTEN/PI3K/AKT signaling pathway / 中国肿瘤生物治疗杂志

@#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.

Gramicidin inhibits human gastric cancer cell proliferation, cell cycle and induced apoptosis

BACKGROUND: Gastric cancer is a common malignant tumor with high morbidity and mortality worldwide, which seriously affects human health. Gramicidin is a short peptide antibiotic which could be used for treating infection induced by bacteria or fungi. However, the anti-cancer effect of gramicidin on gastric cancer cells and its underlying mechanism remains largely unknown. RESULTS: Gastric cancer cells SGC-7901, BGC-823 and normal gastric mucosal cells GES-1 were treated with different concentrations of gramicidin respectively. The results of CCK-8 experiment revealed cellular toxicity of gramicidin to cancer cells while cell colony formation assay showed that gramicidin significantly inhibited the proliferation of gastric cancer cells, but had little effect on normal gastric mucosal cells. In addition, the wound healing assay showed that gramicidin inhibited the migration of SGC-7901 cell. Meanwhile, apoptosis and cell cycle analysis revealed that gramicidin induced cell apoptosis with G2/M cell cycle inhibition. Furthermore, western blot analysis demonstrated that gramicidin down-regulated the expression of cyclinD1 and Bcl-2 as well as the FoxO1 phosphorylation. CONCLUSIONS: The current study illustrated the anti-tumor activity of gramicidin on gastric cancer cells, providing a possibility for gramicidin to be applied in clinical practice for the treatment of gastric cancer.

Inhibitory effect of lumbrokinase on proliferation and induction on apoptosis of gastric cancer SGC7901 cells / 吉林大学学报(医学版)

Objective:To investigate the regulatory effect of lumbrukinase (LBK) on the gastric cancer SGC7901cells, and to clarify its mechanism.Methods:The SGC7901cells in the logarithmic growth phase were selected and divided into control group and 2, 4, 8U·mL-1 LBK groups.MTT assay was used to detect the inhibitory rates of proliferation of SGC7901cells in various groups in vitro at different time (24, 48and 72h) .Cell scratch assay was used to detect the migration abilities of the SGC7901cells in vitro in various groups.Flow cytometry was used to determine the apoptotic rates of SGC7901cells and the pencentages of cells at different cell cycles in various groups.The expression levels of Bcl-2, Bax, and caspase-3in the SGC7901cells in various groups were detected by Western blotting method.Results:The results of MTT assay showed that compared with control group, the inhibitory rates of SGC7901cells in different doses of LBK groups after treated for 24, 48and 72hwere increased (P＜0.01) .The cell scratch assay results showed that compared with control group, the migration distances of SGC7901cells in4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) .The flow cytometry results showed that compared with control group, the apoptotic rates of SGC7901cells in 4and 8U·mL-1 LBK groups were increased significantly (P＜0.01) ;the percentages of cells in G1and S phases were decreased (P＜0.01) ;the percentages of cells in G2phase were increased (P＜0.01) .The results of Western blotting method showed that compared with control group, the Bcl-2protein expression level in the SGC7901cells in 8U·mL-1 LBK group was decreased (P＜0.05) ;the Bax and caspase-3protein expression levels were increased (P＜0.05) .Conclusion:LBK can inhibit the proliferation and migration abilities of SGC7901cells in vitro and induce the apoptosis;its mechanism is achieved through the regulation of expression levels of Bcl-2, Bax, and caspase-3proteins.

Up-Regulation of MiR-1915 Inhibits Proliferation, Invasion, and Migration of Helicobacter pylori-Infected Gastric Cancer Cells via Targeting RAGE

PURPOSE: Helicobacter pylori (HP)-infected gastric cancer (GC) is known to be a fatal malignant tumor, but the molecular mechanisms underlying its proliferation, invasion, and migration remain far from being completely understood. Our aim in this study was to explore miR-1915 expression and its molecular mechanisms in regulating proliferation, invasion, and migration of HP-infected GC cells. MATERIALS AND METHODS: Quantitative real-time PCR and western blot analysis were performed to determine miR-1915 and receptor for advanced glycation end product (RAGE) expression in HP-infected GC tissues and gastritis tissues, as well as human gastric mucosal cell line GES-1 and human GC cell lines SGC-7901 and MKN45. CCK8 assay and transwell assay were performed to detect the proliferation, invasion, and migration capabilities. MiR-1915 mimics and miR-1915 inhibitor were transfected into GC cells to determine the target relationship between miR-1915 and RAGE. RESULTS: MiR-1915 was under-expressed, while RAGE was over-expressed in HP-infected GC tissues and GC cells. Over-expressed miR-1915 could attenuate cellular proliferation, invasion, and migration capacities. RAGE was confirmed to be the target gene of miR-1915 by bioinformatics analysis and luciferase reporter assay. Moreover, HP-infected GC cellular proliferation, invasion, and migration were inhibited after treatment with pcDNA-RAGE. CONCLUSION: MiR-1915 exerted tumor-suppressive effects on cellular proliferation, invasion, and migration of HP-infected GC cells via targeting RAGE, which provided an innovative target candidate for treatment of HP-infected GC.

Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901 / 中国实验方剂学杂志

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.

Ginsenoside Rg3 suppresses the formation of vasculogenic mimicry of gastric cancer SGC7901 cells by inhibiting Wnt/β-catenin pathway in vitro / 中国肿瘤生物治疗杂志

@#Objective: To investigate the effects of ginsenoside Rg3 on the formation of vasculogenic mimicry (VM) in gastric cancer cell line SGC7901 and its molecular mechanism. Methods: MTT assay was used to detect the effect of different concentrations of Rg3 on the proliferation of SGC7901 cells. SGC7901 cells were grouped as follows: BML-284 group, XAV-939 group, Rg3 group, Rg3+ BML-284 group and blank group. Transwell chamber assay was used to detect cell invasion and migration; the formation of VM was observed by tube formation assay; the secretion of MMP-9 and MMP2 was detected by ELISA; the mRNA expressions of GSK-3β and Wnt2B were detected by qPCR; the expression of β-Catenin protein in cells was analyzed by WB; and nuclear entry of β-Catenin was examined by Immunofluorescence. Results: Ginsenoside Rg3 inhibited the proliferation of SGC7901 cells in a time- and concentrationdependent manner; compared with the blank group, 40 mg/L Rg3 significantly inhibited the invasion and migration of SGC7901 cells (both P<0.05) and VM formation (P<0.05); in the meanwhile, the expressions of intracellular GSK-3β, Wnt2B mRNA and β-catenin protein, as well as the nuclear entry of β-catenin were significantly inhibited (all P<0.05). The invasion, migration and VM formation of SGC7901 cells in Rg3+BML-284 group were not significantly different from those in the blank group (all P>0.05). Conclusion: Rg3 can inhibit cell invasion, migration and VM formation in SGC7901 cells by inhibiting the activation of Wnt/β-Catenin pathway.