Human Gastric Cancer BGC-823 Cells Research Articles

Preparation of berberine magnetic nanoparticles and their inhibition of human gastric cancer BGC-823 cells

A new drug delivery system loaded with the drug berberine on carboxymethyl chitosan-coated magnetic nanoparticles (Fe3O4@CMCS-BBR) was prepared and characterized through x-ray diffraction, Fourier transform-infrared spectroscopy, a vibrating sample magnetometer, and transmission electron microscopy. By comparing the size of the uncoated nanoparticles (39.26 nm) and the size of the coated magnetite nanoparticles (73.75 nm), it was found that with the CMCS coating on the magnetite nanoparticles, the dispersion of the nanoparticle improved. The optimum pH testing showed a higher drug encapsulation of 51.23% and drug loading of 17.10% at a pH of 5.5 because of the better interaction of the NH3+ group with the negative functional groups of the CMCS. Furthermore, 85.89% of the drug was released within 72 h. The CCK-8 test results showed that Fe3O4@CMCS-BBR magnetic nanocomposites had good biocompatibility with gastric cancer BGC-823 cells and that Fe3O4@CMCS-BBR effectively inhibited the proliferation of cancer cells. The magnetic experimental results showed that Fe3O4@CMCS-BBR had good responsiveness to external magnetic fields and aggregated in the presence of a magnetic field. The results of targeting experiments showed that fluorescein isothiocyanate emitted a strong yellowish fluorescence in cells, which became stronger over time, and the killing effect on cancer cells became greater. The apoptosis results showed that the apoptosis rate induced by the magnetic nanodrug was 54.90%, indicating that the drug had a promoting effect on the apoptosis of BGC-823 cells.

In vitro study on antitumor activity of aurantiamide acetate extracted from Polygonum capitatum

Aurantiamide acetate is a dipeptide derivative with antibacterial, antiviral, anti-inflammatory and analgesic activities. However, its antitumor activity has not been documented. This study aimed to prepare aurantiamide acetate from Polygonum capitatum and investigate its antitumor activity in vitro. The petroleum ether extract fraction of 80% ethanol extract from P. capitatum was separated by silica gel column chromatography. Aurantiamide acetate was obtained and identified by physicochemical properties, MS and NMR spectral data. The inhibitory effect of different doses of this compound on the proliferation of human umbilical vein endothelial cells (HUVECs), human gastric cancer BGC-823 cells, lung cancer A549 cells and colorectal cancer Caco-2 cells was detected by CCK-8 method. Morphological changes of BGC-823 cells were observed under the inverted microscope. The effect of aurantiamide acetate on BGC-823 apoptosis and cycle was detected by flow cytometry. The expression of Bax and caspase-3 in BGC-823 cells was detected by Western blotting. Aurantiamide acetate inhibited BGC-823, A549 and Caco-2 cell proliferation at the concentrations from 25 μmol/L to 100 μmol/L, but had no inhibitory effect on HUVECs at the concentrations from 12.5 μmol/L to 100 μmol/L. The inhibitory effect of Aurantiamide acetate was most pronounced on BGC-823 cell proliferation at IC50 of 56.73 μmol/L, most probably by increasing the protein expression of caspase-3 and Bax in BGC-823 cells, and arresting cell cycle in G2/M phase. This is the first time that we isolated aurantiamide acetate from P. capitatum with a purity of 98.21%, and found that Aurantiamide acetate could in inhibit the proliferation of various cancer cell types in vitro. This finding may provide a new resource for developing anti-tumor drugs in future.

Synthesis, cytotoxicity, antioxidant activity and molecular modeling of new NSAIDs-EBS derivatives

Two series of NSAIDs-EBS derivatives (5a-j and 9a-i) based on the hybridization of nonsteroidal anti-inflammatory drugs (NSAIDs) skeleton and Ebselen moiety were synthesized. Their cytotoxicity was evaluated against five types of human cancer cell lines, BGC-823 (human gastric cancer cell line), SW480 (human colon adenocarcinoma cells), MCF-7 (human breast adenocarcinoma cells), HeLa (human cervical cancer cells), A549 (human lung carcinoma cells). Moreover, the most active compound 5j showed IC50 values below 3 μM in all cancer cell lines and with remarkable anticancer activity against MCF-7 (1.5 μM) and HeLa (1.7 μM). The redox properties of the NSAIDs-EBS derivatives prepared herein were conducted by 2, 2-didiphenyl-1-picrylhydrazyl (DPPH), bleomycin dependent DNA damage and glutathione peroxidase (GPx)-like assays. Finally, TrxR1 inhibition activity assay and molecular docking study revealed NSAIDs-EBS derivatives could serve as potential TrxR1 inhibitor.

Retracted: Inhibition of Proliferation of SGC7901 and BGC823 Human Gastric Cancer Cells by Ursolic Acid Occurs Through a Caspase-Dependent Apoptotic Pathway.

This publication has been retracted by the Editor due to concerns regarding the originality of the figure images. Reference: Jing Zhang, Fengjun Liu, Xin Zhang. Inhibition of Proliferation of SGC7901 and BGC823 Human Gastric Cancer Cells by Ursolic Acid Occurs Through a Caspase-Dependent Apoptotic Pathway. Med Sci Monit, 2019; 25: 6846-6854. DOI: 10.12659/MSM.916740.

Effect of lentivirus-mediated peroxiredoxins 6 gene silencing on the phenotype of human gastric cancer BGC-823 cells.

Peroxiredoxins (PRDX6) regulates the occurrence and progression of cancer. The aim of this study is to investigate the effect of PRDX6 knockdown on the biological behavior of human gastric cancer cell line BGC-823 cells. Research article. The differential expression of PRDX6 in gastric cancer and normal gastric tissues was tested by immunohistochemistry. Ribonucleic acid plasmid of PRDX6 gene was packaged using a lentivirus, and BGC-823 cells were transfected with the lentivirus to obtain a BGC-823 cell line in which the expression of PRDX6 was stably silenced. The proliferation activity of BGC-823 cells was detected using the cell counting kit-8 method. The effect of PRDX6 on the migration and invasion of BGC-823 cells was evaluated using the scratch test and Transwell assay, and the expression of related proteins was detected by western blot. The expression of PRDX6 in gastric cancer was significantly increased (P < 0.05). Compared with those in the untransfected and negative control groups. The proliferation, migration, and invasion of gastric cancer BGC-823 cells were significantly inhibited, and the apoptotic rates were significantly increased in the lentivirus-transfected (short hairpin-PRDX6) group. Western blot analysis showed that the expression of Bax protein increased, whereas that of proliferating cell nuclear antigen, Bcl-2, PI3K, phospho (p-Akt), and phosphorylated-mammalian target of rapamycin (mTOR) decreased significantly compared with that in WT and vector groups (P < 0.05). The knockdown of PRDX6 gene expression in BGC-823 cells can inhibit the proliferation, migration, and invasion of gastric cancer cells and promote apoptosis, thereby affecting gastric cancer cells.

New organoselenides (NSAIDs-Se derivatives) as potential anticancer agents: Synthesis, biological evaluation and in silico calculations.

Herein we reported the synthesis of twenty new organoselenium compounds (2a-2j and 3a-3j) based on the hybridization of nonsteroidal antiinflammatory drugs (NSAIDs) skeleton and organoselenium motif (-SeCN and -SeCF3), the anticancer activity was evaluated against four types of cancer cell lines, Caco-2 (human colon adenocarcinoma cells), BGC-823 (human gastric cancer cells), MCF-7 (human breast adenocarcinoma cells), PC-3 (human prostatic cancer cells). Interestingly, the introduction of the -SeCN or -SeCF3 moiety in corresponding parent NSAIDs results in the significant effect on cancer cell lines. Moreover, the most active compound 3a showed IC50 values lower than 5μM against the four cancer cell lines, particularly to BGC-823 and MCF-7 with IC50 values of 2.5 and 2.7μM, respectively. Furthermore, three compounds 3a, 3g and 3i were selected to investigate their ability to induce apoptosis in BGC-823cells via modulating the expression of anti-apoptotic Bcl-2 protein, pro-inflammatory cytokines (IL-2) and proapoptotic caspase-8 protein. The redox properties of the NSAIDs-Se derivatives prepared herein were conducted by 2, 2-didiphenyl-1-picrylhydrazyl (DPPH), bleomycin dependent DNA damage and glutathione peroxidase (GPx)-like assays. Finally, molecular docking study revealed that an interaction with the active site of thioredoxin reductase 1 (TrxR1) and predicted the anticancer activity of the synthesized candidates. Overall, these results could serve a promising launch point for further design of NSAIDs-Se derivatives as potential anticancer agents.

3,3'-diindolylmethane exerts antiproliferation and apoptosis induction by TRAF2-p38 axis in gastric cancer.

3,3'-diindolylmethane (DIM), an active phytochemical derivative extracted from cruciferous vegetables, possesses anticancer effects. However, the underlying anticancer mechanism of DIM in gastric cancer remains unknown. Tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2), one of the signal transduction proteins, plays critical role in proliferation and apoptosis of human gastric cancer cells, but there are still lack of practical pharmacological modulators for potential clinical application. Here, we further explored the role of TRAF2 in inhibiting cell proliferation and inducing apoptosis by DIM in human gastric cancer BGC-823 and SGC-7901 cells. After treating BGC-823 and SGC-7901 cells with DIM for 24 h, cell proliferation, apoptosis and TRAF2-related protein were measured. Our findings showed that DIM inhibited the expressions of TRAF2, activated p-p38 and its downstream protein p-p53, which were paralleled with DIM-triggered cells proliferation, inhibition and apoptosis induction. These effects of DIM were reversed by TRAF2 overexpression or p38 mitogen-activated protein kinase (MAPK)-specific inhibitor (SB203580). Taken together, our data suggest that regulating TRAF2/p38 MAPK signaling pathway is essential for inhibiting gastric cancer proliferation and inducing apoptosis by DIM. These findings broaden the understanding of the pharmacological mechanism of DIM's action as a new modulator of TRAF2, and provide a new therapeutic target for human gastric cancer.

Chromodomain-helicase-DNA-binding protein 1-like (CHD1L) silencing inhibits gastric cancer cell proliferation, invasion, and migration.

BackgroundWe aimed to investigate the effect of chromodomain-helicase-DNA-binding protein 1-like (CHD1L) silencing on the biological behavior of gastric cancer cells.MethodsSmall hairpin RNA (shRNAs) targeting CHD1L were designed and transduced into BGC-823 human gastric cancer cells. Expression of p53, p21, nerve growth factor IB (Nur77), and ARHGEF9 was assessed by western blotting, and the effect of CHD1L silencing on gastric cancer cell proliferation, apoptosis, and migration was examined by MTT, flow cytometry, and wound healing assays, respectively.ResultsIn CHD1L-shRNA-1-treated cells, the expression of p53, p21, Nur77, and ARHGEF9 was significantly upregulated, and the number of apoptotic cells was significantly increased compared to the shRNA-negative control (NC; P<0.05). Additionally, the number of cells in the G1 phase was significantly increased among CHD1L-shRNA-1-treated cells. In contrast, the number of cells in the S phase was significantly decreased among CHD1L-shRNA-1-treated cells compared to shRNA-NC-treated cells (P<0.05). Following CHD1L silencing, there were 58.63±10.97 invading cells compared to 144.95±12.68 and 148.49±17.86 in the shRNA-NC and untreated groups, respectively (P<0.05). After 24 h, CHD1L-silenced BGC-823 cells migrated 0.54±0.34 µm compared to 1.34±0.26 and 1.31±0.31 µm in the shRNA-NC and untreated groups, respectively (P<0.05).ConclusionsCHD1L silencing significantly inhibited the proliferation, invasion, and migration of BGC-823 gastric cancer cells and induced apoptosis. Knockdown of CHD1L may present a novel approach for treating gastric cancer.

Effects of miR-141-3p on proliferation, migration and NF-κB signaling pathways in gastric cancer cells

Objective: To investigate the effects of miR-141-3p on proliferation and migration of gastric cancer cells and nuclear factor-κB (NF-κB) signaling pathway. Methods: Human gastric cancer cell line BGC-823 was cultured, and miR-141-3p mimetic (miR-141-3p mimics) was transfected into BGC-823 cells by lipofection. The miR-141-3p overexpressed BGC was constructed. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the transfection effect. The proliferation of BGC-823 cells was determined by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Transwell assay was used to detect the effect of miR-141-3p on BGC-823 cell migration. The expressions of NF-κB p65, p-IKK-α and p-IKB-α protein in NF-κB signaling pathway were detected by western blot. Results: Compared with the control group and the negative control group, the expression level of miR-141-3p in BGC-823 cells of the miR-141-3p group was (2.39±0.27), which was higher than (1.00±0.09) of the control group and (1.01±0.10) of the negative control group (P<0.05). The number of migrating cells in the miR-141-3p group was (47.64±5.65), which was lower than (106.22±12.14) in the control group and (110.40±12.26) in the negative control group (P<0.05). The expression levels of NF-κB p65, p-IKK-α and p-IKB-α protein in BGC-823 cells were down-regulated (P<0.05). Conclusion: MiR-141-3p can inhibit the proliferation and migration of human gastric cancer BGC-823 cells, which may be related to the inhibition of NF-κB signaling pathway activation.

Silencing Stat3 inhibits viability and induces apoptosis in BGC-823 human gastric cancer cell line

ABSTRACT Gastric cancer (GC) is characterized by infrequent early diagnosis, poor prognosis and high mortality. Signal transducer and activator of transcription 3 (Stat3) and signal transducer and activator of transcription 5b (Stat5b) play multiple roles in the development and progression of many human cancers. We investigated the effects of silencing Stat3 and Stat5b on the viability and apoptosis of the human gastric cancer cell line, BGC-823. We found that Stat3 and Stat5b were expressed in both the nucleus and cytoplasm of BGC-823 cells. Silencing of Stat3 caused significantly decreased viability and increased apoptosis, as well as attenuated B-cell lymphoma-2 (Bcl-2) expression in BGC-823 cells. Silencing of Stat5b, however, had no significant effect on these events. Stat3, but not Stat5b, plays an important role in the viability and apoptosis of human gastric cancer cell line, BGC-823, which suggests that Stat3 is a potential target for gastric cancer therapy.

Evaluation of 111In-DOTA-F56 peptide targeting VEGFR1 for potential non-invasive gastric cancer xenografted tumor mice Micro-SPECT imaging

Non-invasive imaging of vascular endothelial growth factor receptor 1 (VEGFR1) remains a great challenge in the early diagnosis of tumors, especially in gastric cancer. Here, we designed and evaluated a novel 111In-DOTA-F56 peptide as a radioactive analogue of F56 (peptide WHSDMEWWYLLG) to bind VEGFR1. It was obtained by radiolabeling DOTA-F56 with 111InCl3 with 98% radiochemical purity and 1.4 ± 0.4 GBq/µmol specific activity. 111In-DOTA-F56 was obtained by the reaction of DOTA-F56 (10 µg) with 111InCl3 in pH 4.0 sodium acetate buffer at 85 °C for 20 min. 111In-DOTA-F56 shows good stability in 0.01 M Phosphate Buffered Saline (PBS) and 5% Human Serum Albumin (HSA). 111In-DOTA-F56 has a high binding affinity for human gastric cancer BGC-823 cells. Bio-distribution studies of 111In-DOTA-F56 were performed in nude mice xenografted with human gastric cancer BGC-823 cells and the results revealed tumor uptake accumulation. A blocking dose of DOTA-F56 significantly reduced the tumor uptake of 111In-DOTA-F56. Tumors were observed with Micro-SPECT images, and the uptake in the tumor increased with time from 4 h to 24 h. The MIP of the Micro-SPECT also showed that the excess DOTA-F56 can specifically block 111In-DOTA-F56 in a mouse tumor model. We successfully synthesized the 111In-DOTA-F56 VEGFR1-targeted peptide as a non-invasive molecule with fine radiochemical properties. Micro-SPECT indicates tumor uptake, which can be further blocked by excess of the F56 peptide, indicating that 111In-DOTA-F56 peptide has potential for early detection of VEGFR1 positive gastric cancer and is worthy of further clinical investigations.

Aberrant methylation of secreted protein acidic and rich in cysteine gene and its significance in gastric cancer.

BACKGROUNDAberrant methylation in DNA regulatory regions could downregulate tumor suppressor genes without changing the sequences. However, our knowledge of secreted protein acidic and rich in cysteine (SPARC) and its aberrant methylation in gastric cancer (GC) is still inadequate. In the present research, we performed fundamental research to clarify the precise function of methylation on SPARC and its significance in GC.AIMTo investigate promoter methylation and the effects of the SPARC gene in GC cells and tissues and to evaluate its clinical significance.METHODSPlasmids that overexpressed the SPARC gene were transfected into human GC BGC-823 cells; non-transfected cells were used as a control group (NC group). Quantitative real-time polymerase chain reaction and western blotting (WB) were then used to detect the expression of SPARC. Methylation-specific polymerase chain reaction was executed to analyze the gene promoter methylation status. Cell viability was measured by the cell counting kit-8 assay. The migration and invasion ability of cells were detected by scratch assays and transwell chamber assays, respectively. Cell cycle events and apoptosis were observed with a flow cytometer.RESULTSThe expression of SPARC mRNA in GC tissues and cells was significantly lower and showed differing degrees of hypermethylation, respectively, than that in normal adjacent tissues and control cells. Treatment with 5-Aza-2’-deoxycytidine (5-Aza-Cdr) was able to restore the expression of SPARC and reverse promoter hypermethylation. Overexpression of the SPARC gene significantly inhibited proliferation, migration, and invasion of GC cells, while also causing cell cycle arrest and apoptosis; the NC group exhibited the opposite effects.CONCLUSIONThis study demonstrated that SPARC could function as a tumor suppressor and might be silenced by promoter hypermethylation. Furthermore, in GC cells, SPARC inhibited migration, invasion, and proliferation, caused cell cycle arrest at the G0/G1 phase, and promoted apoptosis.

Antitumor effects of seleno-short-chain chitosan (SSCC) against human gastric cancer BGC-823 cells.

Seleno-short-chain chitosan (SSCC) is a derivative of chitosan. In the present study, we sought to investigate the underlying antitumor mechanism of SSCC on human gastric cancer BGC-823 cells in vitro. MTT assay suggested that SSCC exhibited a dose-dependent inhibitory effect on the proliferation of BGC-823 cells. We found the SSCC-treated cells showed typical morphological characteristics of apoptosis in a dose dependent manner by observing on microscope. Annexin V-FITC/PI double staining and cell cycle assay identified that SSCC could induce BGC-823 cells apoptosis by triggering G2/M phase arrest. Our research provided the first evidence that SSCC could effectively induce the apoptosis of BGC-823 cells via an intrinsic mitochondrial pathway, as indicated by inducing the disruption of mitochondrial membrane potential (MMP), the excessive accumulation of reactive oxidative species (ROS), the increase of Bax/Bcl-2 ratio and the activation of caspase 3, caspase 9 and cytochrome C (Cyt-C) in BGC-823 cells. These combined results clearly indicated that SSCC could induce BGC-823 cells apoptosis by the involvement of mitochondrial signaling pathway, which provided precise experimental evidence for SSCC as a potential agent in the prevention and treatment of human gastric cancer.

Inhibition of Proliferation of SGC7901 and BGC823 Human Gastric Cancer Cells by Ursolic Acid Occurs Through a Caspase-Dependent Apoptotic Pathway.

BackgroundWorldwide, gastric cancer is one of the most common malignant tumors. Ursolic acid is a plant metabolite and pentacyclic triterpenoid used in traditional Chinese medicine. This study aimed to investigate the effects of ursolic acid the growth and apoptosis of SGC7901 and BGC823 human gastric cancer cells in vitro.Material/MethodsSGC7901 and BGC823 human gastric cancer cells and normal GES-1 gastric epithelial cells were cultured with increasing doses of ursolic acid at 50, 60, and 100 μM. Cell viability and proliferation were assessed using an MTT assay. Flow cytometry was used to assess cell apoptosis. Western blot was used to measure procaspase-8, procaspase-9, procaspase-3, and cleaved poly (ADP-ribose) polymerase (PARP) expression. The expression of receptor interaction protein 3 (RIP3) was examined by Western blot and reverse transcription polymerase chain reaction (RT-PCR). Morphological changes in the gastric cancer cells were determined using Hoechst 33342 staining following ursolic acid treatment.ResultsUrsolic acid inhibited the viability of SGC7901 and BGC823 cells but not GES-1 cells. Ursolic acid treatment significantly induced apoptosis in SGC7901 and BGC823 cells when compared with GES-1 cells (P<0.05), and significantly increased the activation of caspase-3, caspase-8, caspase-9, poly ADPribose polymerase (PARP), and the production of reactive oxygen species (ROS). Treatment of SGC7901 and BGC823 cells with ursolic acid for 72 h did not induce necroptosis.ConclusionsUrsolic acid inhibited the proliferation of SGC7901 and BGC823 human gastric cancer cells in vitro through a caspase-dependent apoptotic pathway.

Changes of the stability and bioactivity of quercetin and myricetin in BGC-823 cells in response to heat treatment and Fe2+/Cu2+ addition

Two flavonols quercetin and myricetin with heat treatment and Fe2+/Cu2+ addition were assessed for stability and bioactivity changes. Quercetin and myricetin at 37 °C showed higher degradation rate constants than those at 20 °C (0.133 − 1.025 vs. 0.299 − 1.803 h−1). Due to the chelation of flavonol and metals, Fe/Cu addition at the two temperatures (except Cu addition at 37 °C) mostly led to enhanced stability via decreasing the constant values to 0.064 − 0.675 h−1. Cell counting kit-8 assaying results showed that myricetin and quercetin heated at 37 °C and 100 °C received decreased growth inhibition in human gastric cancer BGC-823 cells, because they at 80 μmol/L showed decreased inhibition percentages (from 41.1 − 45.6 to 23.9 − 43.3%, cell treatment of 24 h; or from 46.6 − 51.9 to 28.8 − 46.5%, cell treatment of 48 h). The assaying results of Hoechst 33,258staining, DCFH-DA fluorescence probe, and flow cytometry also proved that the heated flavonols in the cells had less reactive oxygen species generation, lower DNA damage, and weakened apoptosis induction, compared with the unheated flavonols. Fe addition decreased these flavonol bioactivities in the cells because of decreased flavonol–Fe chelation and enhanced flavonol degradation; however, Cu addition induced oxidative stress and thus led to less bioactivity decrease than Fe addition. Correlation analysis results indicated that decreased reactive oxygen species generation of the two flavonols was correlated with decreased apoptosis induction (p < 0.05). Overall, these used treatments brought about stability and bioactivity changes for the two flavonols, suggesting reasonable consideration should be paid on processing conditions and some components of foods when studying food bioactivities.

Triterpenoid Saponins from Tu Jia Ethnomedicine Bai San Qi and Their Cytotoxicity on Hep G2 and BGC-823 Cell Lines

ObjectiveTu Jia ethnomedicine is a unique medical system inherited and adhibited by Tu Jia minority living in central-south China. Panax japonicus C. A. Mey. (Bai San Qi, 白三七) is recognized as an effective and rare medicinal plant to treat weakness, fatigue and rheumatism in Tu Jia ethnomedicine. This paper is to discover more substance evidence for the application of Tu Jia ethnomedicine. MethodsColumn chromatography and preparative high performance liquid chromatography (HPLC) was applied for isolation and purification; 1H-NMR, 13C-NMR, 1H-1H COSY, HSQC and HMBC NMR spectra were applied for structure identification; Methyl thiazolyl tetrazolim (MTT) assays were applied for cytotoxicity evaluation. ResultsTotally 12 known compounds were isolated by column chromatography and preparative HPLC from rhizomes of Panax japonicus C. A. Mey. (Bai San Qi, 白三七). Structures of these compounds were identified by their NMR spectra. All the 12 compounds were triterpenoid saponins. Five of them were oleanolic acid type, and the remaining 7 were dammarane type. Eleven compounds were assayed for their cytotoxic activity against Hep G2 human liver cancer cell lines and BGC-823 human gastric cancer cell lines. Three of the 11 showed relatively dominant cytotoxicity against these cell lines. ConclusionsA total of 12 known compounds have been identified from Panax japonicus C. A. Mey. (Bai San Qi, 白三七); NMR spectra of compounds with similar skeletons showed regular characteristics; 3 compounds showed relatively dominant cytotoxicity against Hep G2 and BGC-823 cancer cell lines, and the result can be valued as weak while setting the taxol as a positive control.

Effect of Cu/Mn-Fortification on In Vitro Activities of the Peptic Hydrolysate of Bovine Lactoferrin against Human Gastric Cancer BGC-823 Cells.

Bovine lactoferrin hydrolysate (BLH) was prepared with pepsin, fortified with Cu2+ (Mn2+) 0.64 and 1.28 (0.28 and 0.56) mg/g protein, and then assessed for their activity against human gastric cancer BGC-823 cells. BLH and the four fortified BLH products dose- and time-dependently had growth inhibition on the cells in both short- and long-time experiments. These samples at dose level of 25 mg/mL could stop cell-cycle progression at the G0/G1-phase, damage mitochondrial membrane, and induce cell apoptosis. In total, the fortified BLH products had higher activities in the cells than BLH alone. Moreover, higher Cu/Mn fortification level brought higher effects, and Mn was more effective than Cu to increase these effects. In the treated cells, the apoptosis-related proteins such as Bad, Bax, p53, cytochrome c, caspase-3, and caspase-9 were up-regulated, while Bcl-2 was down-regulated. Caspase-3 activation was also evidenced using a caspase-3 inhibitor, z-VAD-fmk. Thus, Cu- and especially Mn-fortification of BLH brought health benefits such as increased anti-cancer activity in the BGC-823 cells via activating the apoptosis-related proteins to induce cell apoptosis.

Pumilaside A from Litchi semen induces apoptosis in human gastric cancer BGC823 cells via activation of death receptor- and mitochondria-mediated apoptotic pathways

Purpose: To investigate the cytotoxic effect of pumilaside A from Litchi semen against human gastric cancer BGC823 cells, and unravel its possible mechanism(s) of action.Methods: The cytotoxic activity of pumilaside A (5 - 40 μg/mL) against BGC823 cells was assessed by thiazolyl blue tetrazolium bromide assay. The pro-apoptotic effect of PA (10, 20 or 40 μg/mL) on BGC823 cells was monitored by flow cytometry, while the mechanisms involved were investigated using western blot.Results: Pumilaside A significantly produced cytotoxic activity against BGC823 cells (IC50 = 25.43 μg/mL) and induced apoptosis in BGC823 cells (p < 0.01). Treatment with pumilaside A led to significant upregulation of pro-apoptotic factors (Fas, FasL, FADD, Bax, Apaf-1, and c-caspase -8, 9 and 10), and downregulation of anti-apoptotic factors (survivin and Bcl-2, p < 0.05, 0.01). In addition, pumilaside A increased the cytoplasmic levels of Smac and cytochrome c in BGC823 cells by enhancing their mitochondrial release, and significantly upregulated the levels of executioner c-caspases-3, 6 and 7 (p < 0.05, 0.01).Conclusion: Pumilaside A shows good cytotoxic activity against BGC823 cells via a mechanism related to activation of death receptor- and mitochondria-mediated apoptotic pathways. Thus, pumilaside A has a potential for use as an anti-gastric cancer agent.Keywords: Litchi semen, Pumilaside A, BGC823 cells, Cytotoxicity, Apoptosis

Pepsin-catalyzed plastein reaction with tryptophan increases the in vitro activity of lactoferrin hydrolysates with BGC-823 cells

Lactoferrin hydrolysates (LF-H) were digested using pepsin and modified using a pepsin-catalyzed plastein reaction with tryptophan (Trp) to obtain a target product. LF-H, the target product, plastein reaction-modified LF-H alone, and a mixture of LF-H and Trp were evaluated for their activities with human gastric cancer BGC-823 cells at 24 and 48 h. Assay results indicated that the target product had decreased free amino groups and had incorporated Trp. LF-H and the plastein reaction-modified LF-H alone did not show detectable growth inhibition on BGC-823 cells at 1–3 mg/mL; however, the target product at the same dose levels inhibited cell growth, and showed higher activity than the mixture (P < 0.05). The target product at 2 and 3 mg/mL stopped cell cycle progression at the G0/G1-phase, induced DNA fragmentation, and showed apoptosis induction, which increased the number of apoptotic cells. Moreover, time- and dose-dependent responses were observed in these experiments. RT-PCR assay results indicated that exposure of the cells to the target product resulted in up- and down-regulated mRNA expression for 7 apoptosis-related genes, namely Bcl-2, Bcl-xl, Bad, Bax, caspase-3, caspase-6, and caspase-9. Thus, the plastein reaction increased in vitro anticancer activities of LF-H with BGC-823 cells, and more importantly, both the plastein reaction and Trp incorporation contributed to enhanced activities.

Inhibition of the hypoxia-induced factor-1α and vascular endothelial growth factor expression through ginsenoside Rg3 in human gastric cancer cells.

The aim of this study is to probe in the inhibitory effects of ginsenoside Rg3 on the expression of hypoxia-induced factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in human gastric cancer cells. Human gastric cancer BGC823 cells were divided into the control group and experiment group, and expression levels of HIF-1α and VEGF were detected by immunocytochemistry and Western blot after cells were cultured under hypoxia for different durations. Under hypoxia, expression of HIF-1α and VEGF in human gastric cancer BGC823 cells showed an increasing trend, and that was remarkably lower in experiment group than in the control group after applying Rg3, which was obvious at 12 and 24 h (P < 0.05). Rg3 can inhibit expression of HIF-1α and VEGF in human gastric cancer cells and may influence abdominal implantation metastasis of gastric cancer through inhibiting its expression.