Previously undescribed tetrahydroanthraquinones from Prismatomeris tetrandra (roxb.) K. Schum with antitumor cell proliferation activities.

To find structurally previously undescribed compounds with pharmacological effects from Prismatomeris tetrandra (Roxb.) K. Schum (Rubiaceae), thirteen undescribed tetrahydroanthraquinones (1⎼13) named prisconnatanones J⎼V and seven known anthraquinones (14⎼20) were isolated and characterized. The structures of these compounds were elucidated by detailed spectroscopic analyses, and their absolute configurations were established by modified Mosher's method and ECD calculations. The antitumor cell proliferative activities of prisconnatanones J⎼V were determined. Among them, prisconnatanones J possessed high antitumor cell proliferation in HGC27 cells (IC50, 0.792 µM) by blocking HGC27 cells in the S phase and significantly inducing apoptosis in HGC27 cells. Prisconnatanone J has no cytotoxicity to normal gastric cells line (GES-1) at 10 µM and showed a considerable selectivity for HGC27 cells. Prisconnatanone J can potentially inhibit tumor cell proliferation and should be further investigated.

Yiwei decoction promotes apoptosis of gastric cancer cells through spleen-derived exosomes.

Yiwei decoction (YWD) is a formula of traditional Chinese medicine (TCM) that is clinically effective for the prevention and treatment of gastric cancer recurrence and metastasis. According to the theory of TCM, YWD tonifies the body and strengthens the body's resistance to gastric cancer recurrence and metastasis potentially via the immune regulation of the spleen. The aims of the present study were to investigate whether YWD-treated spleen-derived exosomes in rats inhibit the proliferation of tumor cells, to elucidate the anticancer effects of YWD, and to provide evidence supporting the use of YWD as a new clinical treatment for gastric cancer. Spleen-derived exosomes were obtained by ultracentrifugation and identified by transmission electron microscopy, nanoparticle tracking analysis, and western blot analysis. The location of the exosomes in tumor cells was then determined by immunofluorescence staining. After tumor cells were treated with different concentrations of exosomes, the effect of exosomes on cell proliferation was determined by cell counting kit 8 (CCK8) and colony formation assays. Tumor cell apoptosis was detected by flow cytometry. Particle analysis and western blot analysis identified the material extracted from spleen tissue supernatant as exosomes. Immunofluorescence staining showed that spleen-derived exosomes were taken up by HGC-27 cells, and the CCK8 assay confirmed that the relative tumor inhibition rate of YWD-treated spleen-derived exosomes in the 30 µg/mL reached 70.78% compared to control exosomes in the 30 µg/mL (p < 0.05). Compared to control exosomes in the 30 µg/mL, the colony formation assay indicated that YWD-treated spleen-derived exosomes in the 30 µg/mL colonies have decreased by 99.03% (p < 0.01). Moreover, flow cytometry analysis showed that treatment with YWD-treated exosomes in the 30 µg/mL increased the apoptosis rate to 43.27%, which was significantly higher than that of the control group in the 30 µg/mL (25.91%) (p < 0.05). In conclusion, spleen-derived exosomes from YWD-treated animals inhibit the proliferation of HGC-27 cells via inducing apoptosis, suggesting that spleen-derived exosomes are involved in mediating the antitumor effect of YWD. These results demonstrated a novel exosome-mediated anticancer effect of YWD as a TCM formula, thereby supporting the use of YWD-treated exosomes as a new approach for the clinical treatment of gastric cancer.

Isoprenylated Flavonoids and 2-Arylbenzofurans from the Root Bark of Morus alba L. and Their Cytotoxic Activity against HGC27 Cancer Cells.

Three new compounds (1, 11, and 12), together with 32 known ones, were isolated from the root bark of Morus alba L. using various chromatographic methods. The structures of the undescribed compounds were elucidated based on 1D, 2D NMR, and HRESIMS dataanalysis, while the known ones were identified by comparison of their spectroscopic data with those reported in the literature. All the isolates were evaluated for their cytotoxic activities against human gastric cancer HGC27 cells by CCK-8 assay. Among them, compounds 5, 8, 10, and 30 exhibited cytotoxic activities on HGC27 cells with IC50 values of 33.76 ± 2.64 µM, 28.94 ± 0.72 µM, 6.08 ± 0.34 µM, and 10.24 ± 0.89 µM, respectively. Furthermore, compound 10 was confirmed to reduce proliferation ability, increase apoptosis rate, and inhibit cell migration pathway by annexin V/PI double staining experiment, transwell experiment, and Western blot analysis.

Ginkgolide B inhibits the malignant biological behaviors of gastric cancer HGC-27 cells by blocking the PI3K/Akt/mTOR signaling pathway / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨银杏内酯B（GKB）是否通过阻抑PI3K/Akt/mTOR信号通路抑制胃癌HGC-27细胞的增殖、凋亡、迁移及侵袭。方法：将HGC-27细胞分为对照、GKB低剂量（100 mg/L）、GKB高剂量（200 mg/L）、GKB高剂量（200 mg/L）+740Y-P（PI3K激活剂）、Ly294002（PI3K抑制剂）组。采用MTT、Edu、FCM、Transwell实验分别检测各组细胞的增殖、凋亡、迁移和侵袭能力，qPCR和WB法分别检测各组细胞中PI3K mRNA、Akt mRNA、mTOR mRNA和Ki-67、caspase-3、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白的表达。构建胃癌HGC-27细胞裸鼠移植瘤模型，观察GKB对移植瘤生长的影响，WB法检测移植瘤组织中Ki-67、caspase-3、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白的表达。结果：体外实验结果表明，与对照组相比，GKB低剂量组、GKB高剂量组、Ly294002组HGC-27细胞的增殖活力及细胞增殖率、迁移和侵袭细胞数，PI3K、Akt、mTOR mRNA表达，以及Ki-67、p-PI3K/PI3K、p-Akt/Akt、p-mTOR/mTOR蛋白表达均显著降低（均P<0.05）；细胞凋亡率、caspase-3蛋白表达均显著升高（均P<0.05）；740Y-P可部分逆转GKB对HGC-27细胞的抑制作用（均P<0.05）。荷瘤裸鼠实验结果显示，GKB可显著抑制HGC-27细胞裸鼠移植瘤的生长（P<0.05），且可下调PI3K/Akt/mTOR通路相关蛋白的表达。结论：GKB可通过阻抑PI3K/Akt/mTOR信号通路而抑制胃癌HGC-27细胞增殖、迁移与侵袭并促进其凋亡。

Design, Synthesis and Biological Evaluation of Neocryptolepine Derivatives as Potential Anti-Gastric Cancer Agents.

Natural products play an important role in drug development and lead compound synthesis. Neocryptolepine is a polycyclic quinoline compound isolated from Cryptolepis sanguinolent. The cytotoxicity of neocryptolepine to gastric cancer cells AGS, MKN45, HGC27, and SGC7901 was not very strong, and it also had certain toxicity to gastric mucosa cells GES-1. Therefore, a series of neocryptolepine derivatives were synthesized by the modification of the structure of neocryptolepine, and their cytotoxicity was evaluated. The results showed that compounds C5 and C8 exhibited strong cytotoxicity to AGS cells. The cell colony formation and cell migration experiments suggested that compounds C5 and C8 could inhibit the proliferation and cell migration of AGS and HGC27 cells. Cell cycle and apoptosis experiments showed that compounds C5 and C8 did not cause the apoptosis of AGS and HGC27 cells but, mainly, caused cell necrosis. Compound C5 had no significant effect on AGS and HGC27 cell cycles at low concentration. After treatment with AGS cells for 24 h at high concentration, compound C5 could significantly arrest the AGS cell cycle in the G2/M phase. Compound C8 had no significant effect on the AGS and HGC27 cell cycles. The results of molecular docking and Western blot showed that compounds C5 and C8 might induce cytotoxicity through the PI3K/AKT signaling pathway. Therefore, compounds C5 and C8 may be promising lead compounds for the treatment of gastric cancer.

Daphnane-Type Diterpenes from Stelleropsis tianschanica and Their Antitumor Activity.

Four new daphnane-type diterpenes named tianchaterpenes C-F (1-4) and six known ones were isolated from Stelleropsis tianschanica. Their structures were elucidated based on chemical and spectral analyses. The comparisons of calculated and experimental electronic circular dichroism (ECD) methods were used to determine the absolute configurations of new compounds. Additionally, compounds 1-10 were evaluated for their cytotoxic activities against HGC-27 cell lines; the results demonstrate that compound 2 had strong cytotoxic activities with IC50 values of 8.8 µM, for which activity was better than that of cisplatin (13.2 ± 0.67 µM).

Investigation of the anticancer and apoptotic effect of Micromeria congesta under in vitro conditions and detection of related genes by real-time PCR.

At the present time cancer is one of the biggest health problems and because of the problems encountered in its treatment, alternative treatment methods of herbal origin are researched. In this study, the cytotoxic effects of the essential oil extracted from the Micromeria congesta plant on various cancer cells (A549, ECC-1, HCT-116, HELA, HGC-27, MDA-MB-231, SNU-423, U20S, DLD-1, PC-3) and normal cells (BEAS-2B, CRL-4010) have been examined. Anticancer mechanism of action has been particularly examined on gastric cancer (HGC-27; IC50: 15.84 µg mL-1), on which essential oil showed a high cytotoxic effect. In the study, the cytotoxic effect and the apoptotic effect have been applied by MTT and flow cytometric annexin-V methods, respectively. The apoptotic gene expression (caspase 3, caspase 9, MMP2, MMP9, ACTB) real-time PCR content analysis has been performed with gas chromatography mass spectrometry (GC-MS). M. congesta essentials oil has the highest cytotoxic effect on gastric cancer (HGC-27) cells, decreases MMP2 and MMP9 expressions, and induces apoptosis with increasing the expression of caspase 3 and caspase 8 genes. In addition, it has been determined that piperitenone oxide (40.00 - 45.00%), pulegone (11.00%) and cyclohexanone (18.00 - 19.00%) are the major components of M. congesta essentials oil. In conclusion, it has been determined that the compounds found in high amounts in M. congesta plant induces apoptosis by affecting the expression of compound genes and thus can have the potential to be an alternative drug in the treatment of gastric cancer.

Design, Synthesis and Bioactivity Evaluation of Novel Chalcone Derivatives Possessing Tryptophan Moiety with Dual Activities of Anti-Cancer and Partially Restoring the Proliferation of Normal Kidney Cells Pre-Treated with Cisplatin.

BACKGROUND: Chalcone is a broad-spectrum natural product with anti-cancer and anti-inflammatory activities. However, low potency, low selectivity, and serious side effects limit its druggability. L-Tryptophan is an essential precursor molecule of an anti-cancer active substance. Also, the indole moiety inhibits the proliferation of tumor cells by binding to colchicine sites. A decrease in kidney cell activity caused by kidney inflammation is the primary side effect of cancer therapy. OBJECTIVE: The purpose of this work was to design, synthesize, and perform bioactivity evaluation of novel chalcone derivatives possessing tryptophan moiety with dual activities of anti-cancer and partially restoring the proliferation of normal kidney cells pre-treated with cisplatin. METHODS: A series of novel chalcone derivatives possessing tryptophan moiety (5a-5g, 6a-6o) were designed, synthesized, and evaluated for anti-cancer activity against four cancer cell lines (gastric (HGC-27), colon (HCT-116), prostate (PC-3), and lung (A549)), and a human normal cell line (gastric mucosal epithelial (GES-1)). The activity of restoring the proliferation of normal kidney cells pre-treated with cisplatin was evaluated by MTT assay. Cell cycle, apoptosis, and apoptosis proteins (Bax and Bcl-2) were used to evaluate the anti-cancer mechanism of the most potent compound. Moreover, a docking study was performed to explain the high anti-cancer activity of 6n. The expressions of TNF-α, IL- 6, and MCP-1 were detected by ELISA. RESULTS: Most of the compounds exhibited high anti-cancer activity against the HGC-27 cell line and exhibited low toxicity against the normal cell line. Based on three rounds of a structure optimization, 6n was discovered as the most potent compound against HGC-27 cells with an IC50 value of 2.02 µM and an SI value of 28.47. Further studies demonstrated that 6n could induce cell cycle arrest at the G2/M phase and the apoptosis of the HGC-27 cell line by reducing the expression of Bcl-2 and improving the expression level of Bax. Molecular docking result displayed 6n bound to the colchicine site. At the same time, 6n also exhibited moderate activity of restoring the proliferation of normal kidney cells pre-treated with cisplatin by reducing the expression of inflammatory substances. CONCLUSION: Our findings collectively suggested that 6n should be further studied as a potential anti-cancer agent that could partially restore the proliferation of normal kidney cells pre-treated with cisplatin in gastric cancer patients by an anti-inflammatory pathway.

Inhibitory Effect of Hederasaponin B on Gastric Cancer HGC-27 Cells and Its Mechanism Based on Bioinformatics / 中国实验方剂学杂志

ObjectiveTo investigate the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell and the mechanism. MethodMethyl thiazolyl tetrazolium （MTT） assay， hematoxylin-eosin （HE） staining， 4'，6-diamidino-2-phenylindote （DAPI） staining， colony formation assay， scratch assay， and flow cytometry were employed for the analysis of apoptosis and cell cycle. Thereby， the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell was investigated. Then the Pharm Mapper， UniProt， Swissdock， STRING， and Metascape were used for target screening， gene annotation， molecular docking， protein-protein interaction （PPI） network construction， Gene Ontology （GO） term and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis to explore the mechanism. ResultHederasaponin B （15， 30， 60， 120 μmol·L-1） can significantly reduce the survival rate of HGC-27 cell （P<0.01） in a time-dependent and dose-dependent manner compared with the blank group. It had no significant toxicity to normal GES-1 cell at concentration below 120 μmol·L-1. Compared with the blank group， hederasaponin B （30， 60， 120 μmol·L-1） induced cytoplasmic vacuolization， and nuclear deformation and karyopyknosis， inhibited the migration of HGC-27 cell （P<0.01）， and brought about the apoptosis （P<0.05， P<0.01） and cell cycle arrest of HGC-27 cell （P<0.05， P<0.01）. Hederasaponin B （10， 20， 30 μmol·L-1） also suppressed the independent survival ability and proliferation ability of HGC-27 cell （P<0.01）. The possible action targets were kinesin-like protein KIF11， cGMP-specific 3，5 cyclic phosphodiesterase， caspase-3， serine/threonine protein kinase Chk1， proto-oncogene tyrosine protein kinase， epidermal growth factor receptor， and mitogen-activated protein kinase （MAPK） 8. The mechanism may be related to MAPK signaling pathway （pathways in cancer）， adhesion connection， focal adhesion and proteoglycans in cancer （epithelial cell signaling pathways in Helicobacter pylori infection）. ConclusionHederasaponin B exerts significant inhibitory effect on gastric cancer HGC-27 cell through multiple targets and multiple pathways.

Expression of mitogen-activated protein kinase kinase 1 in gastric cancer and its clinical significance / 解剖学报

Objective To investigate the expression of mitogen-activated protein kinase kinase 1 (MAP2K1) in gastric cancer and its clinical significance. Methods Immunohistochemistry and Western blotting were used to detect the protein expression of MAP2K1 in gastric cancertissues and cells. The morphology and the expression position of MAP2K1 were observed by immunofluorescence. MAP2K1 mRNA expression in gastric cancer tissues was analyzed by data mining of Starbase database and Oneomine database. The correlation between MAP2K1 mRNA expression and clinicopathological features was analyzed by UALCAN database. Survival analysis was performed using Kaplan Meier-Plotter online analysis tools. GEPIA2 database mining the relationship between MAP2K1 and gastric cancer stem cell related factors and drug resistance related factors. Results Immunohistochemistry, immunofluorescence and Western blotting showed that MAP2K1 protein was highly expressed in gastric cancer tissues and cells, and MAP2K1 was expressed in the cytoplasm of gastric cancer. According to the analysis of various databases, the expression of MAP2K1 mRNA in gastric cancer tissue was higher than that in normal gastric tissue, and the expression of MAP2K1 mRNA was closely related to gastric cancer stage, grade, lymph node metastasis and patient gender, and the overall survival rate of gastric cancer patients in the group with high MAP2K1 mRNA expression was significantly lower than that in the group with low MAP2K1 mRNA expression, which may be related to the characteristics of gastric cancer stem cells and drug resistance. Conclusion MAP2K1 is highly expressed in gastric cancer, and its expression level may affect the poor prognosis of patients by regulating stem cell related factors and drug resistance related factors. MAP2K1 may be a new diagnostic marker to determine the prognosis of gastric cancer patients.

Direct interaction between Rab5a and Rab4a enhanced epidermal growth factor-stimulated proliferation of gastric cancer cells.

BACKGROUND: Gastric cancer (GC) is one of the leading causes of cancer-related death worldwide. Although targeted therapies such as antibodies against human epidermal growth factor receptor 2 or vascular endothelial growth factor receptor 2 have been widely used in the treatment of metastatic cancer, the overall outcomes are poor. Therefore, elucidation of the mechanism underlying cancer progression is important to improve prognosis. Overexpression of the Rab5a gene has been confirmed to correlate with tumorigenesis of many cancers, but the mechanism underling, especially of GC, is still unclear. AIM: To investigate the effects of Rab5a overexpression on the tumorigenesis of GC. METHODS: First, the expression levels of Rab5a and Rab4a in primary tumorous tissues of GC patients diagnosed between 2015 and 2018 were analyzed. Then we constructed HGC-27 cell lines overexpressing green fluorescent protein-Rab5a or red fluorescent protein-Rab4a and investigated the interaction between Rab5a or Rab4a using Western blotting, co-immunoprecipitation, confocal microscopy, and colocalization analysis. Finally, epidermal growth factor-stimulated proliferation of these cell lines was analyzed using cell counting kit-8 cell viability assay. RESULTS: Compared with normal gastric tissues, the expression levels of Rab5a and Rab4a increased progressively both in paracancerous tissues and in advanced cancerous tissues. Epidermal growth factor could promote the proliferation of HGC-27 cells, especially Rab5a-overexpressing HGC-27 cells. Notably, Rab5a and Rab4a co-overexpression promoted the proliferation of HGC-27 cells to the greatest extent. Further analysis identified a direct interaction between Rab5a and Rab4a in HGC-27 cells. CONCLUSION: Co-overexpression of Rab5a and Rab4a in GC may promote the endosomal recycling of epidermal growth factor receptor, which in turn contributes to poor prognosis and tumor progression in GC patients. Inhibition of Rab5a or Rab4a expression might be a promising therapy for refractory GC.

CircTMCO3 Promotes Gastric Cancer Progression by Regulating miR-577/RAB14 Axis.

BACKGROUND: Growing evidence indicated that circRNAs played major roles in the progression of human cancer. Nevertheless, the molecular mechanism and effects of circTMCO3 in GC are still unclear. METHODS: First, qRT-PCR was used to evaluate the levels of circTMCO3 from GC tissues, GC cells, normal tissues and gastric epithelial cells. Then, the GC cells were transfected to analyze the proliferation, migration and invasion of GC cells by MTT, colony formation and transwell assays. Next, the expressions of miR-577 and RAB14 in GC tissues and cells were examined by qRT-PCR following transfection. The target interaction of circTMCO3-miR-577 and miR-577-RAB14 was explored by the dual-luciferase reporter and RNA pull-down assays. In the end, the growth and viability of GC cells were detected by MTT, colony formation and transwell assays, respectively, following the transfection of GC cells. RESULTS: In this research, we found circTMCO3 expressions are significantly up-regulated in GC tissues and cells compared with the normal tissues and gastric epithelial cells. We discovered that the knockdown of circTMCO3 remarkably inhibits the proliferation, migration and invasion of GC cells. Besides, through the prediction of binding sites between circTMCO3, miR-577 and RAB14, we discovered miR-577 is a target of circTMCO3 while RAB14 is a target gene of miR-577. Finally, the results demonstrate the overexpression of miR-577 and the silence of RAB14 could inhibit the effects of circTMCO3 on proliferation, migration and invasion in GC cells. CONCLUSION: circTMCO3 accelerated the growth and migration of GC cells by regulating miR-577/RAB14 axis.

Apatinib inhibits gastric carcinoma development by regulating the expression levels of IL-17 via the Bax/Bcl-2 signaling pathway.

Gastric carcinoma is a common type of gastrointestinal tumor with high morbidity and mortality rates. IL-17 is a newly discovered cytokine that has been reported to serve an important role in the development of gastric carcinoma. The potential effect of apatinib on IL-17 expression levels in the development of gastric carcinoma has been rarely reported. The present study aimed to investigate the potential mechanism of IL-17 and apatinib in the development of gastric carcinoma. A total of 30 tumor and para-carcinoma tissues were collected from 30 patients with gastric carcinoma between January 2019 and December 2019 and the expression levels of IL-17 in the tissues were analyzed by reverse transcription-quantitative PCR and western blotting. An in vitro model of gastric carcinoma was also established using the HGC-27 cell line, in which the cells were divided into control, IL-17, IL-17-apatinib and apatinib groups. The expression levels of IL-17, Bax, Bcl-2 and caspase-3 were analyzed using reverse transcription-quantitative PCR and western blotting. An MTT assay and flow cytometry were used to analyze the proliferation and apoptosis of HGC-27 cells, respectively, and a Transwell assay was used to analyze the invasive ability of HGC-27 cells. The results revealed that the expression levels of IL-17 were significantly upregulated in the gastric carcinoma tissues compared with the para-carcinoma tissues. In vitro, IL-17 treatment promoted the proliferation and invasive ability of HGC-27 cells, but inhibited the apoptosis with the significantly downregulated expression levels of Bax and caspase-3 and the upregulated expression levels of Bcl-2 than control group. Conversely, apatinib treatment significantly inhibited the proliferative and invasive abilities of HGC-27 cells, but promoted cell apoptosis in the IL-17 and IL-17-apatinib groups.. Collectively, the present results suggested that the upregulation of IL-17 may be associated with the occurrence and development of gastric carcinoma. The findings indicated that apatinib may inhibit gastric carcinoma development by regulating IL-17 expression via the Bax/Bcl-2 signaling pathway. Therefore, the present findings may enhance the current knowledge of the effect of apatinib on gastric carcinoma cells.

LINC01018 regulates the proliferation, apoptosis and radiosensitivity of gastric cancer HGC-27 cells through miR-297 / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨长基因间非编码RNA（long intergene non-coding RNA，LINC）01018是否通过抑制miR-297调控胃癌HGC-27细胞的增殖、凋亡及放射敏感性。方法： 收集于青海省第五人民医院接受手术的胃癌患者（21例）及化疗抵抗胃癌患者（19例）的癌组织和癌旁组织，以qPCR检测胃癌组织、化疗抵抗胃癌组织和胃癌HGC-27细胞中LINC01018、miR-297的表达。双荧光素酶报告基因实验验证LINC01018与miR-297之间的靶向关系。在HGC-27细胞中转染LINC01018过表达质粒pcDNA-LINC01018或miR-297抑制剂，或共转染pcDNA-LINC01018和miR-297模拟物，以qPCR检测验证细胞转染效果。MTT和克隆形成实验检测转染后HGC-27细胞的增殖活力与克隆形成能力，流式细胞术检测转染后HGC-27细胞的凋亡率，克隆形成实验检测各组转染后HGC-27细胞的放射敏感性，WB实验检测细胞中Ki67、cleaved-caspase3、pro-caspase3蛋白的表达。结果： 与癌旁组织或胃正常黏膜上皮细胞GES-1相比，胃癌组织、化疗抵抗胃癌组织和胃癌HGC-27细胞中LINC01018呈低表达、miR-297呈高表达（均P<0.01）。LINC01018与miR-297存在靶向结合关系，LINC01018负向调控miR-297的表达。过表达LINC01018或敲减miR-297可抑制HGC-27细胞的增殖活力与克隆形成能力、降低细胞存活率，促进细胞的凋亡、下调Ki67和pro-caspase3蛋白表达水平、上调cleaved-caspase3蛋白表达水平、提高HGC-27细胞的放射敏感性（均P<0.01）。共转染pcDNA-LINC01018和miR-297模拟物可逆转过表达LINC01018对HGC-27细胞的所有上述作用，尤其可逆转其对HGC-27细胞的放射增敏作用（P<0.05或P<0.01）。结论： LINC01018通过下调miR-297表达抑制胃癌HGC-27细胞增殖而促进凋亡和增强细胞的放射敏感性，该作用与Ki67和caspase3的表达变化有关。

Effects of microRNA-4286 on the growth, migration and invasion of gastric cancer cell line HGC-27 by regulating inositol polyphosphate-4-phosphatase type I / 解剖学报

Objective To investigate the effects of microRNA-4286 (miRNA-4286) on the growth, migration and invasion of gastric cancer cell line HGC-27 by regulating inositol polyphosphate-4-phosphatase type I (INPP4A). Methods HGC-27 cells were divided into blank control group, negative control group and intervention group. The blank control group received no treatment, negative control group was transfected with pEGFP-Nl plasmid, and intervention group was transfected with pEGFP-Nl-miRNA-4286 plasmid. Real-time PCR was used to detect the expression of miRNA-4286 in HGC-27 cells, and Western blotting was used to detect the expression of INPP4A protein in HGC-27 cells. The proliferation rate, invasion ability and migration ability of HGC-27 cells were detected by cholecystokinin octapeptide, cholecystokinin octapeptide (CCK8), Transwell chamber assay and scratch test, respectively. Results The miRNA-4286 expression level, proliferation rate, and number of transmembrane cells in intervention group were significantly higher than those in blank control group and negative control group (P0.05). Conclusion miRNA-4286 may promote the growth, migration and invasion of gastric cancer cell line HGC-27 by down- regulating the expression of INPP4A.

Genetic Profiles Affect the Biological Effects of Serine on Gastric Cancer Cells.

A high serine content in body fluid was identified in a portion of patients with gastric cancer, but its biological significance was not clear. Here, we investigated the biological effect of serine on gastric cancer cells. Serine was added into the culture medium of MGC803 and HGC27 cancer cells, and its influence on multiple biological functions, such as cell growth, migration and invasion, and drug resistance was analyzed. We examined the global transcriptomic profiles in these cultured cells with high serine content. Both MGC803 and HGC27 cell lines were originated from male patients, however, their basal gene expression patterns were very different. The finding of cell differentiation-associated genes, ALPI, KRT18, TM4SF1, KRT81, A2M, MT1E, MUC16, BASP1, TUSC3, and PRSS21 in MGC803 cells suggested that this cell line was more poorly differentiated, compared to HGC27 cell line. When the serine concentration was increased to 150mg/ml in medium, the response of these two gastric cancer cell lines was different, particularly on cell growth, cell migration, and invasion and 5-FU resistance. In animal experiment, administration of high concentration of serine promoted cancer cell metastasis to local lymph node. Taken together, we characterized the basal gene expressing profiles of MGC803 and HGC27. The HGC27 cells were more differentiated than MGC803 cells. MGC803 cells were more sensitive to the change of serine content. Our results suggested that the responsiveness of cancer cells to microenvironmental change is associated with their genetic background.

Effect of simulated microgravity on metabolism of HGC-27 gastric cancer cells.

The understanding into the pathogenesis and treatment of gastric cancer has improved in recent years; however, a number of limitations have delayed the development of effective treatment. Cancer cells can undergo glycolysis and inhibit oxidative phosphorylation in the presence of oxygen (Warburg effect). Previous studies have demonstrated that a rotary cell culture system (RCCS) can induce glycolytic metabolism. In addition, the potential of regulating cancer cells by targeting their metabolites has led to the rapid development of metabolomics. In the present study, human HGC-27 gastric cancer cells were cultured in a RCCS bioreactor, simulating weightlessness. Subsequently, liquid chromatography-mass spectrometry was used to examine the effects of simulated microgravity (SMG) on the metabolism of HGC-27 cells. A total of 67 differentially regulated metabolites were identified, including upregulated and downregulated metabolites. Compared with the normal gravity group, phosphatidyl ethanolamine, phosphatidyl choline, arachidonic acid and sphinganine were significantly upregulated in SMG conditions, whereas sphingomyelin, phosphatidyl serine, phosphatidic acid, L-proline, creatine, pantothenic acid, oxidized glutathione, adenosine diphosphate and adenosine triphosphate were significantly downregulated. The Human Metabolome Database compound analysis revealed that lipids and lipid-like metabolites were primarily affected in an SMG environment in the present study. Overall, the findings of the present study may aid our understanding of gastric cancer by identifying the underlying mechanisms of metabolism of the disease under SMG.

The Volatilomic Footprints of Human HGC-27 and CLS-145 Gastric Cancer Cell Lines.

The presence of certain volatile biomarkers in the breath of patients with gastric cancer has been reported by several studies; however, the origin of these compounds remains controversial. In vitro studies, involving gastric cancer cells may address this problem and aid in revealing the biochemical pathways underlying the production and metabolism of gastric cancer volatile indicators. Gas chromatography with mass spectrometric detection, coupled with headspace needle trap extraction as the pre-concentration technique, has been applied to map the volatilomic footprints of human HGC-27 and CLS-145 gastric cancer cell lines and normal Human Stomach Epithelial Cells (HSEC). In total, 27 volatile compounds are found to be associated with metabolism occurring in HGC-27, CLS-145, and HSEC. Amongst these, the headspace concentrations of 12 volatiles were found to be reduced compared to those above just the cultivating medium, namely there was an observed uptake of eight aldehydes (2-methylpropanal, 2-methyl-2-propenal, 2-methylbutanal, 3-methylbutanal, hexanal, heptanal, nonanal, and benzaldehyde), three heterocyclic compounds (2-methyl-furan, 2-ethyl-furan, and 2-pentyl-furan), and one sulfur-containing compound (dimethyl disulphide). For the other 15 volatiles, the headspace concentrations above the healthy and cancerous cells were found to be higher than those found above the cultivating medium, namely the cells were found to release three esters (ethyl acetate, ethyl propanoate, and ethyl 2-methylbutyrate), seven ketones (2-pentanone, 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, 2-pentadecanone, and 2-heptadecanone), three alcohols (2-methyl-1-butanol, 3-methyl-1-butanol, and 2-ethyl-1-hexanol), one aromatic compound (toluene), and one sulfur containing compound [2-methyl-5-(methylthio) furan]. In comparison to HSEC, HGC-27 cancer cell lines were found to have significantly altered metabolism, manifested by an increased production of methyl ketones containing an odd number of carbons. Amongst these species, three volatiles were found exclusively to be produced by this cell line, namely 2-undecanone, 2-tridecanone, and 2-heptadecanone. Another interesting feature of the HGC-27 footprint is the lowered level of alcohols and esters. The CLS-145 cells exhibited less pronounced changes in their volatilomic pattern compared to HSEC. Their footprint was characterized by the upregulated production of esters and 2-ethyl-hexanol and downregulated production of other alcohols. We have therefore demonstrated that it is possible to differentiate between cancerous and healthy gastric cells using biochemical volatile signatures.

Cytotoxicity of lanostane-type triterpenoids and ergosteroids isolated from Omphalia lapidescens on MDA-MB-231 and HGC-27 cells.

Omphalia lapidescens Schroet. is a medicinal macrofungus in China that has shown good antitumor activity in recent research. To explore the potential cytotoxic compounds from O. lapidescens, the ethyl acetate soluble fraction of a 95% ethanol extract with cytotoxic activity was phytochemically investigated. A new tetranorlanostane triterpenoid (1) and a new ergosteroid (4), together with two known lanostanes (2 and 3) and six known Δ22-ergosteroids (5-10), were isolated. Notably, compound 1 possesses a new 24,25,26,27-tetranorlanostane carbon skeleton. All isolates, except compound 8, were tested for cytotoxic activities using a human breast cancer cell line (MDA-MB-231) and a human gastric cancer cell line (HGC-27) in vitro. The new nortriterpenoid (1) exhibited no obvious cytotoxicity, while the known triterpenoid (3) showed significant cytotoxicity against MDA-MB-231 and HGC-27 cells. Compound 4 exhibited the highest cytotoxicity against MDA-MB-231 and HGC-27 cells, with IC50 values of 11.33 ±â€¯2.18 and 12.28 ±â€¯3.64 µM, respectively. Furthermore, Hoechst fluorescence 33342 staining and Western blot tests demonstrated that compound 4 induced apoptosis in MDA-MB-231 cells by downregulating procaspase-3 expression and upregulating the expression ratio of Bax/Bcl-2.

Voltage-dependent calcium channel α2δ1 subunit is a specific candidate marker for identifying gastric cancer stem cells.

Purpose: Cancer stem cells (CSCs) are a subpopulation of cancer cells with self-renewal property and responsible for tumor malignancy, progression and drug resistance. Researches on CSC-specific markers in gastric cancer remain limited. Our current study explored the expression of voltage-dependent calcium channel α2δ1 subunit and the potential of using α2δ1 as a CSC marker in gastric cancer. We also compared the specificity of α2δ1 and CD44 in identifying gastric cancer stem cells (GCSCs). Materials and methods: Expression of α2δ1 was analyzed in gastric cancer cell lines, patient-derived xenograft (PDX) models and clinical samples of malignant ascites of gastric cancer patients. α2δ1+ gastric cancer cells were isolated from gastric cancer cell lines. CSC properties of α2δ1+ gastric cancer cells were then verified by subsequent tests both in vitro and in vivo. Results: The expression level of α2δ1 was found to differ drastically among gastric cancer cell lines, PDX models and clinical samples of malignant ascites. α2δ1+ gastric cancer cells sorted from HGC-27 and SGC-7901 cell lines demonstrated significant self-renewal properties, including tumorigenic capacity, sphere-formation capacity and asymmetric differentiation potential. Knockdown of α2δ1 in α2δ1+ HGC-27 significantly inhibited CSC properties and rendered HGC-27 more sensitive to chemotherapy. Flow cytometry showed that α2δ1+ gastric cancer cells accounted for a small fraction of CD44+ gastric cancer cells. Isolated CD44+α2δ1+ HGC-27 cells displayed more significant tumorigenic capacity and sphere-forming capacity compared with their CD44+α2δ1- counterparts. Conclusion: α2δ1+ gastric cancer cells possessed CSC properties. α2δ1 could be a proper marker in identifying GCSCs with superior specificity than CD44. The combination of α2δ1 and CD44 could be used to identify GCSCs with improved accuracy. Knockdown of α2δ1 combined with chemotherapy displayed higher therapeutic efficacy on gastric cancer cells, suggesting that α2δ1 could be a potential target for anticancer treatment.