Gastric Cancer Cell Lines AGS Research Articles

Mg-doped ZnO nanoparticle as an effective nanocarrier in delivery of 5-Fluorouracil anti-gastric cancer drug

5-Fluorouracil (5-FU) is a chemotherapy drug that belongs to a class of medications known as antimetabolites and has many applications in cancer therapy, especially in the treatment of gastric cancer. However, high dose of 5-FU in gastric cancer treatment can cause various side effects. Therefore, delivery of this significant drug with a suitable delivery system can significantly improve the safety and efficacy of this drug, leading to better treatment outcomes and improved patient quality of life. Mg-doped ZnO has garnered significant attention in scientific research due to its distinct physicochemical characteristics. This investigation introduces an uncomplicated and economical approach for synthesis of Mg-doped ZnO nanoparticles. Then, the nanoparticles were subjected to thorough analysis employing FT-IR, XRD, XPS, FE-SEM, TEM, UV–Vis, EDS, and DLS techniques. Subsequently, the research explored capacity of the created nanostructure to serve as a nanocarrier for 5-Fluorouracil. Findings indicated an approximately 76 % drug loading capacity that is dependent on pH and duration time. The release profile depicted a gradual release under physiological conditions (pH 7.4, 39 %), contrasting with a notably accelerated release rate at pH 5.3 (82 %) in 48 h. Furthermore, an in vitro assessment of cellular cytotoxicity performed on AGS gastric cancer cell line to evaluate safety and effectiveness of Mg-doped ZnO nanoparticles. Results from MTT assay affirmed the potential of this nanoparticle as a promising vehicle for 5-Fluorouracil delivery.

Extraction and Isolation of Two Polysaccharides from Chloranthus japonicus Sieb. and Evaluation of Their Anti-Gastric Cancer Activities.

Two unreported heteropolysaccharides, denoted as YCJP-1 and YCJP-2, were isolated from the herbs of Chloranthus japonicus. YCJP-1 was a heteropolysaccharide composed of glucose, galactose, arabinose, mannose, rhamnose, and a minor proportion of uronic acids, with the molecular weight mainly distributed in the 74,475-228,443 Da range. YCJP-2 was mainly composed of glucose, mannose, and galactose, with the molecular weights ranging from 848 to 5810 Da. To further evaluate the anti-gastric cancer effects of C. japonicus, the inhibitory effects of the crude polysaccharide (YCJP) and the purified polysaccharides (YCJP-1 and YCJP-2) were determined using a CCK-8 assay and colon-forming assay on MGC-803 and AGS gastric cancer cell lines. Our results showed that YCJP, YCJP-1, and YCJP-2 possess prominent inhibitory effects on the proliferation of MGC-803 and AGS cells, and the AGS cell was more sensitive to YCJP, YCJP-1, and YCJP-2. Moreover, YCJP-2 demonstrated superior anti-gastric cancer effects compared to YCJP-1. This could potentially be attributed to YCJP-2's higher glucose content and narrower molecular weight distribution.

O247: In vitro exposure of gastric cancer cell line AGS to the cannabinoid cannabidiol (CBD) results in cytotoxicity in a dose-dependent manner

Abstract Introduction Current curative therapeutic regimens for gastric cancer combine chemotherapy with surgery, though this approach is associated with morbidity and mortality and relatively low long-term survival when compared to other solid organ and gastro-intestinal tumours. Cannabinoids are known to have anti-cancer properties in some tumours, though their role in gastric cancer is relatively unknown. This study aimed to determine if the non-psychoactive cannabidiol (CBD) induces cytotoxicity in the gastric cancer cell line AGS. Method The gastric cancer cell line AGS was maintained as per recommended guidelines. When cells reached 70% confluence, they were seeded in 96 well plates and treated with CBD or cisplatin at concentrations ranging from 0.2 μM to 100 μM or vehicle control. After 96 hours contact time, cell viability was assessed using a colorimetric cell viability assay (MTT assay). Experiments were repeated independently four times. Cell viability at each concentration was recorded and half maximal inhibitory concentration (IC50) was calculated. Results were expressed as the mean ± SE mean. Results Treatment with cisplatin induced significant cytotoxicity in a dose dependent manner with an IC50 concentration of 1.8 ± 0.3μM. Treatment with CBD also induced cytotoxicity in a dose dependent manner. Cytotoxic effects were observed with concentrations as low as 3.1μM (78% viability). The IC50 concentration for CBD was 10.2 ± 2.5 μM. Conclusions CBD induced significant cytotoxicity at micromolar concentration in stomach carcinoma cells. Further studies should be carried out to investigate the mechanisms of action that underpin the observed in vitro cytotoxic effect.

O248: Treatment of gastric cancer cell line AGS with cannabidiol (CBD) induces loss of mitochondrial membrane potential

Abstract Introduction Cannabinoids have been shown to have anti-cancer properties for a variety of tumour sites; however, the mechanisms that underpin these effects are poorly understood. Apoptosis is implicated as the likely mechanism of action by which cannabinoids induce cell death. The loss of mitochondrial membrane potential (MMP) is observed in apoptotic cells and has been linked to induction of the apoptotic process. This study aimed to assess whether exposure to CBD induced loss of MMP. Method AGS cells were sub-cultured as recommended. When cells reached 70% confluence, they were treated with either control (ethanol), 5 µM, 10 µM or 20 µM CBD. After 24 hours, the MMP assay was performed using the NC-3000 advanced image cytometer to assess the mitochondrial uptake of JC-1 dye. Cells with a loss of mitochondrial membrane potential are represented as percentage of the total cell population and expressed as mean ± SE mean (N=3). The ANOVA test was used to determine statistical significance. Results Treatment with control induced loss of MMP in 8.4 ± 1.7% of cells. Treatment with 5 µM, 10 µM or 20 µM CBD induced a loss of MMP in 6.7 ± 1.4 %, 10.3 ± 2.2% and 44.7 ± 2.8 % of cells respectively. Treatment with 20µM CBD induced a statistically significant loss of MMP when compared to control (p<0.05). Conclusions AGS cells treated with CBD undergo loss of MMP. This finding could indicate that treatment is inducing apoptosis. Further investigations into apoptotic mechanisms are required to confirm the findings.

Identification of ALYREF in pan cancer as a novel cancer prognostic biomarker and potential regulatory mechanism in gastric cancer

ALYREF is considered as a specific mRNA m5C-binding protein which recognizes m5C sites in RNA and facilitates the export of RNA from the nucleus to the cytoplasm. Expressed in various tissues and highly involved in the transcriptional regulation, ALYREF has the potential to become a novel diagnostic marker and therapeutic target for cancer patients. However, few studies focused on its function during carcinogenesis and progress. In order to explore the role of ALYREF on tumorigenesis, TCGA and GTEx databases were used to investigate the relationship of ALYREF to pan-cancer. We found that ALYREF was highly expressed in majority of cancer types and that elevated expression level was positively associated with poor prognosis in many cancers. GO and KEGG analysis showed that ALYREF to be essential in regulating the cell cycle and gene mismatch repair in tumor progression. The correlation analysis of tumor heterogeneity indicated that ALYREF could be specially correlated to the tumor stemness in stomach adenocarcinoma (STAD). Furthermore, we investigate the potential function of ALYREF on gastric carcinogenesis. Prognostic analysis of different molecular subtypes of gastric cancer (GC) unfolded that high ALYREF expression leads to poor prognosis in certain subtypes of GC. Finally, enrichment analysis revealed that ALYREF-related genes possess the function of regulating cell cycle and apoptosis that cause further influences in GC tumor progression. For further verification, we knocked down the expression of ALYREF by siRNA in GC cell line AGS. Knockdown of ALYREF distinctly contributed to inhibition of GC cell proliferation. Moreover, it is observed that knocked-down of ALYREF induced AGS cells arrested in G1 phase and increased cell apoptosis. Our findings highlighted the essential function of ALYREF in tumorigenesis and revealed the specific contribution of ALYREF to gastric carcinogenesis through pan-cancer analysis and biological experiments.

GRB7 plays a promoting role in the progression of gastric cancer

BackgroundGastric cancer is a clinically common tumor, showing an upward trend of both incidence and mortality. GRB7 has been identified as a vital regulator in tumor progression. This study aims to uncover the biological function of GRB7 in gastric cancer process.Methodsimmunohistochemical (IHC) staining using a tissue microarray (TMA), quantitative reverse transcription PCR (qRT-PCR) and Western blotting were performed to detect the expression of genes. Furthermore, gastric cancer cell lines AGS and MGC-803 were transfected with short hairpin RNAs against GRB7. The biological function of GRB7 in gastric cancer cells were examined by CCK-8, flow cytometry, wound healing and Transwell assays. Then, in vivo tumor formation assay was conducted to explore the effects of GRB7 on tumor growth. Finally, expression levels of proteins related to cell functions were determined by Western blotting. Coimmunoprecipitation (CoIP) assay was performed to assess the protein-protein interaction.ResultsGRB7 was up-regulated in gastric cancer tissues and cell lines, and its expression was inversely proportional to survival of gastric cancer patients. Moreover, GRB7 knockdown inhibited proliferative, migratory abilities, as well as promoted cell apoptosis in gastric cancer cells. Further study suggested that GRB7 silencing could suppress gastric cancer tumor growth in vivo. Furthermore, our study uncovered an important interaction between GRB7 and MyD88. Silencing MyD88 was observed to alleviate the malignant phenotypes promoted by GRB7 in gastric cancer cells.ConclusionsTogether, this study provided evidence that GRB7 may be an effective molecular targets for the treatment of gastric cancer.

Methionine restriction attenuates the migration and invasion of gastric cancer cells by inhibiting nuclear p65 translocation through TRIM47.

The prevention and treatment of gastric cancer has been the focus and difficulty of medical research. We aimed to explore the mechanism of inhibiting migration and invasion of gastric cancer cells by methionine restriction (MR). The human gastric cancer cell lines AGS and MKN45 cultured with complete medium (CM) or medium without methionine were used for invitro experiments. MKN45 cells were injected tail vein into BALB/c nude mice and then fed with normal diet or methionine diet for invivo experiments. MR treatment decreased cell migration and invasion, increased E-cadherin expression, decreased N-cadherin and p-p65 expressions, and inhibited nuclear p65 translocation of AGS and MKN45 cells when compared with CM group. MR treatment increased IκBα protein expression and protein stability, and decreased IκBα protein ubiquitination level and TRIM47 expression. TRIM47 interacted with IκBα protein, and overexpression of TRIM47 reversed the regulatory effects of MR. TRIM47 promoted lung metastasis formation and partially attenuated the effect of MR on metastasis formation invivo compared to normal diet group mice. MR reduces TRIM47 expression, leads to the degradation of IκBα, and then inhibits the translocation of nuclear p65 and the migration and invasion of gastric cancer cells.

Metformin reduces the cellular DNA repair capacity and enhances the effect of curcumin on the induction of apoptosis in AGS gastric cancer cells

BackgroundCancer results from the accumulation of mutations in critical genes, such as DNA repair genes. But these genes are a double-edged sword, because the basis of current cancer treatment is DNA damage from chemotherapy and radiation, and the repair system can slow the healing process by repairing the induced damage. Therefore, any substance that can reduce the DNA repair capacity of cancer cells can make the cells more sensitive to treatment. Metformin and curcumin, as low-complication compounds, can play this role well.MethodsIn the present work, changes in the expression of CASP3, BAX, and BCL2L1 apoptotic genes, and nine genes involved in DNA repair pathways (XRCC1, XRCC2, XRCC3, XRCC4, XRCC5, XRCC6, XRCC7, BRCA1 and BRAC2) were measured comparatively by real-time PCR in AGS gastric cancer cell line under single and co-treatments with metformin and curcumin.ResultsOur findings showed that co-treatment of metformin and curcumin induced decreasing the expression of anti-apoptotic BCL2L1 and increasing expression of proapoptotic CASP3 and BAX. Metformin decreased the expression levels of seven genes, while curcumin did not alter the expression levels. The co-treatment of metformin and curcumin showed that although the XRCC2, XRCC3, XRCC4, XRCC5, XRCC6, XRCC7, BRCA1, and BRCA2 were down-regulated, there was no difference between metformin and co-treatment for mRNA levels.ConclusionOur results suggest that metformin increases the sensitivity of cancer cells to anticancer drugs by suppressing several DNA repair pathways and that curcumin may induce apoptosis.

Cancer-associated fibroblasts influence Wnt/PCP signaling in gastric cancer cells by cytoneme-based dissemination of ROR2

Cancer-associated fibroblasts (CAFs) are a crucial component in the tumor microenvironment influencing cancer progression. Besides shaping the extracellular matrix, these fibroblasts provide signaling factors to facilitate tumor survival and alter tumor behavior. In gastric cancer, one crucial signaling pathway influencing invasion and metastasis is the Wnt/Planar Cell Polarity (PCP) signaling. The crucial PCP ligand in this context is WNT5A, which is produced by the CAFs, and gastric cancer cells react upon this signal by enhanced polarized migration. Why gastric cancer cells respond to this signal is still unclear, as their expression level for the central WNT5A receptor, ROR2, is very low. Here, we show that CAFs display long and branched filopodia that form an extensive, complex network engulfing gastric cancer cells, such as the gastric cancer cell line AGS. CAFs have a significantly higher expression level of ROR2 than normal gastric fibroblasts and AGS cells. By high-resolution imaging, we observe a direct transfer of fluorescently tagged ROR2 from CAF to AGS cells by signaling filopodia, known as cytonemes. Surprisingly, we find that the transferred ROR2 complexes can activate Wnt/JNK signaling in AGS cells. Consistently, blockage of ROR2 function in the CAFs leads to reduced paracrine Wnt/JNK signaling, cell polarization, and migration of the receiving AGS cells. Complementary, enhanced migration via paracrine ROR2 transfer was observed in a zebrafish in vivo model. These findings demonstrate a fresh role for cytoneme-mediated signaling in the tumor microenvironment. Cytonemes convey Wnt receptors from CAFs to gastric cancer cells, allowing them to respond to Wnt/PCP signals.

Herbal melanin modulates PGE2 and IL-6 gastroprotective markers through COX-2 and TLR4 signaling in the gastric cancer cell line AGS

We reported a gastric anti-ulcerogenic effect of the Nigella sativa (L.)-derived herbal melanin (HM) using rat models. However, the molecular mechanisms underlying this HM gastroprotective effect remain unknown. Cyclooxygenase-2 (COX-2)-catalyzed prostaglandin E2 (PGE2) and toll-like receptor 4 (TLR4)-mediated interleukin-6 (IL-6) production and secretion play major roles in gastric mucosal protection. In the current study, the human gastric carcinoma epithelial cell line AGS was used as a model to investigate the effect of HM on TLR4, COX-2, glycoprotein mucin 4 protein and gene expression using immuno-cyto-fluorescence staining, Western blot technology, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Gastroprotective markers PGE2 and IL-6 production and secretion were also assessed using an enzyme-linked immunosorbent assay (ELISA). Bacterial lipopolysaccharides (LPS), well-known inducers of TLR4, COX-2, PGE2 and IL-6 expression, were used as a positive control. We showed that HM upregulated its main receptor TLR4 gene and protein expression in AGS cells. HM increased, in a dose- and time-dependent manner, the secretion of PGE2 and the expression of COX-2 mRNA and protein, which was detected in the nucleus, cytoplasm and predominantly at the intercellular junctions of the AGS cells. In addition, HM enhanced IL-6 production and secretion, and upregulated the mucin 4 gene expression, the hallmarks of gastroprotection. To check whether HM-induced PGE2 and IL-6 through TLR4 signaling and COX-2 generated, AGS cells were pre-treated with a TLR4 signaling inhibitor TAK242 and the COX-2 inhibitor NS-398. A loss of the stimulatory effects of HM on COX-2, PGE2 and IL-6 production and secretion was observed in TAK242 and NS-398-pre-treated AGS cells, confirming the role of TLR4 signaling and COX-2 generated in the HM gastroprotective effects. In conclusion, our results showed that HM enhances TLR4/COX-2-mediated secretion of gastroprotective markers PGE2 and IL-6, and upregulates mucin 4 gene expression in the human gastric epithelial cell line AGS, which may contribute to the promising beneficial gastroprotective effect of HM for human gastric prevention and treatment.

Production of the exopolysaccharide from Lactiplantibacillus plantarum YT013 under different growth conditions: optimum parameters and mathematical analysis

ABSTRACT Exopolysaccharides (EPSs) are highly heterogeneous natural polymers, can exhibit some unique bio-logical activities, such as anti-oxidation, inflammation regulation, anticancer and anti-viral activities, etc. A new type of EPS was obtained from the fermentation supernatant of Lactiplantibacillus plantarum YT013. However, the low yield of EPS posed a challenge for further research and application. Therefore, in this study, the production process of EPS was optimized, and anti-cancer cells proliferation activity was investigated. Identified the best single-factor conditions for EPS production, including initial pH 6, inoculation amount 5% (v/v), temperature 37°C, cultivation time 36 h, glucose 3% (w/v), soy peptone 1.5% (w/v), KH2PO4 0.3% (w/v). Under these conditions, the maximum production of EPS reached 0.630 mg/mL, which is 1.31 times higher than that before optimization. Concurrently, the nonlinear model was used for fitting, and it was found that the Voigt nonlinear model was in better agreement with the optimization results, which can be used as an effective tool for optimizing EPS production conditions. Furthermore, the anti-proliferative activity of EPS was evaluated using the human gastric cancer AGS cell line in vitro. MTT assay revealed a strong inhibitory effect of EPS on AGS cell proliferation. Additionally, flow cytometry analysis indicated that EPS promoted apoptosis in AGS cells. Lactiplantibacillus plantarum YT013 EPS, as a biodegradable and nontoxic natural active substance, shows potential as a natural antitumor substance.

Development of allogenic nonviral RNA-based CAR-NK therapy targeting CDH17 in relapsed/refractory gastrointestinal cancer.

e14539 Background: Gastrointestinal (GI) cancers are commonly fatal after metastasis. The incidence and mortality of GI tumors are high, and the benefits of existing therapies are limited. Cadherin-17 (CDH17) is highly prevalent and specific to GI cancers which has potential for the development of novel treatments for GI cancers. Cell-based immunotherapy using genetically engineered autologous Chimeric Antigen Receptors T (CAR-T) cells has been clinically efficacious for treating hematological malignancies. However, its manufacturing is extremely expensive and lengthy with ~10% failure production rate. Equipping NK cells with CAR may represent a safer and equally effective approach for use in solid tumors as an alternative to autologous CAR-T-cell therapy. Nonviral RNA-based expression of CAR vector in NK cells will provide immediate and multiple accessibility to cancer patients, lower risk of abnormal genetic alteration mediated by viral integration, and potentially limit side effects. This paper demonstrates the development of RNA-CAR-NK targeting CDH17-positive GI cancers. Methods: Ex vivo expanded human peripheral blood NK (Magicell-NK, Medigen, Taiwan) cells were used in this study. To generate anti-CDH17 (C7) CAR-NK cells, mRNA containing a novel CDH17-specific single-chain variable fragment and the 4-1BB costimulatory domain was electroporated to Magicell-NK cell. Flow cytometry analysis was performed to evaluate the cell viability and the expression level of CAR and cytotoxic surface markers of the CAR-NK cells. CDH17 specific cytolytic activity against AGS gastric cancer cell line and the concentration of cytokines released by CAR-NK cells were measured by in vitro cytotoxicity assay and cytokine release assay, respectively. Results: Our results demonstrated efficient electroporation with high-level C7 CAR expression ( > 80%) and optimal CAR-NK cell viability (~60%). Interestingly, 12 h co-culture of C7 CAR-NK cells with AGS gastric cancer cells at 1:1 and 2:1 effector/target (E:T) ratio showed a significantly enhanced antitumor response of C7 CAR-NK cells on AGS-CDH17+ cells but not on AGS-CDH17- cells, indicating CDH17 target-specific cytolytic activity. Moreover, we observed an elevated level of cytokines such as INF-γ and TNF-α released by C7 CAR-NK cells only in co-culture with AGS-CDH17+ cells, confirming the mechanisms behind the CAR-mediated cytotoxicity. Furthermore, these results were supported by an increased level of surface CD107a or LAMP-1 expression in C7 CAR-NK cells during the 12 h of co-culture with AGS-CDH17+ cells at 1:1 and 2:1 E:T ratio. Conclusions: Our findings suggest that a non-viral based CDH17 CAR-NK cell therapy could be a potential allogeneic, off-the-shelf CAR-NK therapy candidate in GI cancer and support the rationale of further investigating it in vivo and clinical trials.

Metformin and p-coumaric acid downregulate the expression of hTERT in gastric cancer cell line AGS

Gastric cancer is among the most common and fetal cancers worldwide. Its treatments are expensive and cause side effects. Therefore, treating gastric cancer with natural compounds and known therapeutics, or combinations of them can be interesting alternatives. p-Coumaric acid (pCA) and metformin (Met) are among such anticancer agents. Gastric cancer cells express a high level of human telomerase reverse transcriptase (hTERT) gene, which enhances their invasion and metastasis. As a result, down-regulation of the hTERT gene may be an effective anticancer therapy. Since the effects of treatments with pCA, Met, and their combination on h-TERT expression have not been investigated, the present study was carried out to assess these effects. MTT assay was used to measure the cytotoxic effects of pCA and/or Met on the AGS cells during 48 h. Real-time PCR was used to assess the changes in the expression levels of hTERT gene after 48 h. Wound healing assay, transwell migration and invasion assays, and colony formation assay were performed to determine the metastatic potentials of the AGS cells treated with pCA and/or Met. pCA and Met had dose-dependent cytotoxic effects on the AGS cell line. Treatments with pCA, Met, and their combination significantly suppressed hTERT expression, significantly decreased cellular migration and invasion, and also significantly lowered colony numbers. pCA and Met decrease the expression of hTERT at very low, non-cytotoxic concentrations and inhibit migration, invasion, and colony formation in the AGS gastric cancer cell line. Therefore they may be potential candidates for further investigations in fighting against gastric cancer and may pave the way for the emergence on new anti-gastric cancer therapeutics.

Cytotoxic Activity of Amaryllidaceae Plants against Cancer Cells: Biotechnological, In Vitro, and In Silico Approaches.

Cancer is a major cause of death and an impediment to increasing life expectancy worldwide. With the aim of finding new molecules for chemotherapeutic treatment of epidemiological relevance, ten alkaloid fractions from Amaryllidaceae species were tested against six cancer cell lines (AGS, BT-549, HEC-1B, MCF-7, MDA-MB 231, and PC3) with HaCat as a control cell line. Some species determined as critically endangered with minimal availability were propagated using in vitro plant tissue culture techniques. Molecular docking studies were carried out to illustrate binding orientations of the 30 Amaryllidaceae alkaloids identified in the active site of some molecular targets involved with anti-cancer activity for potential anti-cancer drugs. In gastric cancer cell line AGS, the best results (lower cell viability percentages) were obtained for Crinum jagus (48.06 ± 3.35%) and Eucharis bonplandii (45.79 ± 3.05%) at 30 µg/mL. The research focused on evaluating the identified alkaloids on the Bcl-2 protein family (Mcl-1 and Bcl-xL) and HK2, where the in vitro, in silico and statistical results suggest that powelline and buphanidrine alkaloids could present cytotoxic activity. Finally, combining experimental and theoretical assays allowed us to identify and characterize potentially useful alkaloids for cancer treatment.

Evaluation of the effect of nano-encapsulated lactoferrin on the expression of Bak and Bax genes in gastric cancer cell line AGS and study of the molecular docking of lactoferrin with these proteins

lactoferrin (Lf) is a glycoprotein with various biological activities, including antibacterial, antiviral, anti-cancer, etc. In the present study, the effect of different concentrations of nano-encapsulated lactoferrin (NE-Lf) on the expression of Bax and Bak genes was evaluated in stomach cancer cell line AGS using real-time PCR technique and cytotoxicity of NE-Lf on the growth cells as well as the molecular mechanism of these two genes and their proteins in the apoptosis pathway and the relationship between lactoferrin and these proteins were investigated by bioinformatics studies. In the viability test, the results showed that the growth inhibition effect of nano-lactoferrin was greater than lactoferrin in both concentrations, and chitosan had no inhibitory effect on the cells. In concentrations of 250 and 500 µg of NE-Lf Bax gene expression increased by 2.3 and 5 times, respectively, and Bak gene expression increased by 1.94 and 1.74 times, respectively. Statistical analysis showed that there is a significant difference in the relative amount of gene expression between the treatments in both genes (P < 0.05). The binding mode of lactoferrin with Bax and Bak proteins was obtained using docking. According to docking results, the N-lobe region of lactoferrin interacts with the Bax protein, as well as the Bak protein. The results show that lactoferrin, in addition to acting on the gene, interacts with Bax and Bak proteins. Since two proteins are components of apoptosis, lactoferrin can induce apoptosis in this way.

Lycopene suppresses gastric cancer cell growth without affecting normal gastric epithelial cells

Gastric cancer is one of the leading causes of cancer-related death worldwide. Lycopene, a natural carotenoid, has potent antioxidant activity and anti-cancer effects against several types of cancers. However, the mechanism for the anti-gastric cancer effects of lycopene remains to be fully clarified. Normal gastric epithelial cell line GES-1 and gastric cancer cell line AGS, SGC-7901, Hs746T cells were treated with different concentrations of lycopene and the effects of lycopene were compared. Lycopene specifically suppressed cell growth monitored by Real-Time Cell Analyzer, induced cell cycle arrest and cell apoptosis detected by flow cytometry, and lowered mitochondrial membrane potentials assessed by JC-1 staining of AGS and SGC-7901 cells, while did not affect those of GES-1 cells. Lycopene did not affect the cell growth of Hs746T cells harboring TP53 mutation. Further bioinformatics analysis predicted 57 genes with up-regulated expression levels in gastric cancer and decreased function in cells after lycopene treatment. Quantitative PCR and Western Blot were used to check the critical factors in the cell cycle and apoptosis signaling pathway. Lycopene decreased the high expression levels of CCNE1 and increased the levels of TP53 in AGS and SGC-7901 cells without affecting those in GES-1 cells. In summary, lycopene could effectively suppress gastric cancer cells with CCNE1-amplification, which could be a promising target therapy reagent for gastric cancer.

The effect of potassium-competitive acid blocker tegoprazan on gastric cancer cells.

385 Background: Proton pump inhibitors (PPI) and potassium-competitive acid blocker (P-CAB) are both effective gastric acid secretion blockers. PPI was reported to inhibit proliferation and induce apoptosis of gastric cancer. However, no study on the effect of Tegoprazan, a novel P-CAB, on gastric cancer have been attempted. Methods: In this study, the anti-cancer effect of tegoprazan in gastric cancer cell line AGS and MKN74 were analyzed with esomeprazole (PPI) as a control. After treatment with tegoprazan and esomeprazole, AGS cells were analyzed with with RNA sequencing and Ingenuity Pathway Analysis (IPA). MTT assays, AnnexinV-FITC/PI staining, flow cytometry and western blot were used for subsequent analysis. The wound-healing assay, matrigel chamber assays, colony forming assay, and western blot of epithelial-mesenchymal transition (EMT) markers were used for analysis of oncogenic and migratory ability. Results: In RNA sequencing and subsequent IPA analysis, tegoprazan significantly increased the pathways involved in cell death of gastric cancer cell compared to esomeprazole. MTT assays proved that tegoprazan inhibits proliferation of AGS and MKN74 cells in dose-dependent manner. AnnexinV-FITC/PI staining and western blot demonstrated that tegoprazan stimulates apoptosis of gastric cancer cells. PI staining and flow cytometry confirmed that tegoprazan blocks G1/S phase entry, finally leading to cell cycle arrest. Moreover, wound-healing assay, matrigel chamber assays, colony forming assay, and western blot of epithelial-mesenchymal transition (EMT) markers indicated that tegoprazan inhibits cell migration, invasion, colony formation, and EMT. To determine key molecule of anti-cancer effect of tegoprazan, upstream regulator analysis in IPA was done and cMYC has strong effect on G1/S phase entry, through PI3K/AKT/GSK3β signaling pathway in this experiment. In this study, the experimental results for tegoprazan showed significant changes compared to esomeprazole. Conclusions: Our findings showed that tegoprazan has more evident anti-cancer effect than esomeprazole. cMYC could be a key molecule for anti-cancer effect through PI3K/AKT/GSK3β signaling pathway by tegoprazan in gastric cancer.

Genomic instability-related twelve-microRNA signatures for predicting the prognosis of gastric cancer

Gastric cancer (GC) ranks fifth among all malignant tumors globally, especially in East Asia, and has attracted extensive attention and research. MicroRNA (miRNA) modulation during genomic instability (GI) may be associated with the development and metastasis of malignant tumors. We aimed to identify GI-related miRNA signatures for the prediction of GC prognosis. We constructed a GI-related miRNA signature (GIMiSig) scheme based on The Cancer Genome Atlas (TCGA) training set (n = 389), which was later verified based on the TCGA test set (n = 194). GI-related miRNAs were identified by analyzing somatic mutation profiles and miRNA expression. A GI-related miRNA-gene co-expression network was also constructed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were analyzed to reveal possible biological pathways associated with GI-related miRNAs. The correlation of the GIMiSig with clinical factors of the TCGA dataset was analyzed. MiRNA mimics and inhibitors were used to evaluate the biological functions of miR-100-5p and miR-145-3p in GC cell lines AGS and MKN-45. This study identified a GI-related 12-miRNA signature for the prediction of GC prognosis. GIMiSig scores, similar to tumor stages, showed significant correlations with overall survival (OS, p < 0.05). GIMiSig showed high accuracy in predicting GC prognosis. MiR-100-5p and miR-145-3p promoted cell growth, invasion, and migration but inhibited apoptosis in GC cells. We report a reliable GI-related 12-miRNA signature for predicting GC prognosis. Furthermore, miR-100-5p and miR-145-3p may promote GC cell growth, invasion, and migration.

Green, facile and fast synthesis of silver nanoparticles by using solution plasma techniques and their antibacterial and anticancer activities.

We herein present a simple, fast, efficient and environmentally friendly method for preparing silver nanoparticles (AgNPs) using the solution plasma method in the presence of extracts from Paramignya trimera (P. trimera). The effects of P. trimera extract concentrations and the applied voltage on the formation of AgNPs were investigated. Surface plasmon resonance spectra show a strong peak at 413 nm for the prepared samples. The Fourier-transform infrared spectroscopy measurement results indicated the presence of possible functional groups in the prepared AgNPs. Morphological analysis revealed that the AgNPs were spherical with an average size of 8 nm. The prepared AgNPs exhibited good stability in solution compared to that of AgNPs prepared by the solution plasma technique without P. trimera extract. The formation mechanism of AgNPs is also proposed. The prepared AgNPs exhibited high antibacterial ability against Gram (+) Staphylococcus aureus, Gram (-) Pseudomonas aeruginosa bacteria and strong anticancer activity for the AGS gastric cancer cell line. The obtained results demonstrated that this is a simple, rapid, environmentally friendly method for preparing AgNPs instead of conventional methods using chemical reducing agents for potential applications.

Elettaria Cardamomum Ekstraktının Proleukin İlacı Kombinasyonu ile Mide Kanseri Hücre Hattı Üzerindeki İmmünostimulan/Sitotoksik Etkilerinin İncelenmesi

Kanser, hücrelerin kontrolsüz çoğalması sonucunda meydana gelmektedir. Farklı kanser türleri olmakla birlikte mide kanseri dünyada en çok görülen kanser türleri arasında yer almaktadır. Mide kanserine karşı birçok tedavi yaklaşımı geliştirilmiştir. Ancak günümüzde kullanılan tedavi yöntemleri, kanser hücrelerini öldürmekle birlikte sağlıklı hücrelere de zarar vermektedir. Kullanılan tedavi yöntemlerinin dezavantajlarından dolayı bitkisel tedavi yaklaşımları kullanılmaya başlanmıştır. Elettaria Cardamomum (E. Cardamomum) (kakule) ekstraktında bulunun Diindolilmetan (DIM) ve indol-3-karbinol (I3C) molekülleri hormon yolaklarını baskılayarak kanser hücrelerini öldürdüğü literatürde belirtilmektedir. Bu çalışmada, E. Cardamomum ve proleukin ilaç kombinasyonlarının AGS, J774, THP-1 hücre hatları üzerindeki etkisi araştırılmıştır. Sonuç olarak, kakule ekstraktında bulunan I3C ve DIM molekülleri sayesinde AGS mide kanseri hücre hattında yüksek sitotoksik etkiye sahip olduğu düşünülmektedir. Kakule ekstraktı ile kombine edilen proleukin ilacının ise AGS hücrelerinde kakule ekstraktının tek başına kullanımına göre daha fazla öldürme oranına sahip olduğu belirlenmiştir.