Genome-wide investigation of lncRNAs revealed their tight association with gastric cancer.

BACKGROUND: Gastric cancer (GC) is a significant health issue globally, ranking as the fifth most common cancer with over 10,000 new cases reported annually. Long non-coding RNA (lncRNA) has emerged as a critical player in cellular functions, influencing GC's development, growth, metastasis, and prognosis. However, our understanding of lncRNA's role in the pathogenesis of GC remains limited. Therefore, it is particularly important to explore the relationship between lncRNA and gastric cancer. METHODS: we conducted a comprehensive analysis of RNA sequencing data from the GEO database and stomach adenocarcinoma (STAD) data from the TCGA database to identify lncRNAs that exhibit altered expression levels in GC and the mechanisms underlying lncRNA-mediated transcription and post-transcriptional regulation were explored. RESULTS: This study uncovered 94 lncRNAs with differential expression and, through co-expression analysis, linked these to 1508 differentially expressed genes (DEGs). GO functional enrichment analysis highlighted that these DEGs are involved in critical pathways, such as cell adhesion and the positive regulation of cell migration. By establishing a lncRNA-miRNA-mRNA regulatory network, we found that the ceRNA mechanism, particularly involving RP11-357H14.17 and CTD-2377D24.4, could play a role in GC progression. Experimental validation of selected differentially expressed lncRNAs and mRNAs (including RP11-357H14.17-CLDN1, BBOX1, TRPM2-AS, CLDN1, PLAU, HOXB7) confirmed the RNA-seq results. CONCLUSIONS: Overall, our findings highlight the critical role of the lncRNA-mRNA regulatory network in the development and progression of GC, offering potential biomarkers for diagnosis and targets for innovative treatment strategies.

Differential metabolic networks in three energy substances of flaxseed (Linum usitatissimum L.) during germination.

Germinated flaxseed (Linum usitatissimum L.) is an essential potential food ingredient, but the major energy substances (proteins, lipids, and carbohydrates) metabolites and metabolic pathways are unknown. Comprehensive metabolomic analyses were performed using Fourier transform infrared spectroscopy and high-performance liquid chromatography mass spectrometry on flaxseed from 0 to 7 d. Additionally, the critical metabolites pathways networks of three energy substances metabolites during flaxseed germination were exhibited. The results showed that arginine was the most active metabolite during germination, strongly associated with the arginine biosynthesis and arginine and proline metabolism pathways. Carbohydrates predominantly comprised sucrose on 0-3 d, which participated in galactose metabolism and starch and sucrose metabolism. The main flaxseed phospholipid molecules were phosphatidic acid, phosphatidylethanolamine, lysophosphatidic acid, and lysophosphatidylcholine during germination. This study underscores the paramount metabolic pathways in proteins, lipids and carbohydrates were arginine and proline metabolism, linoleic acid metabolism, arachidonic acid metabolism, and ascorbate and aldarate metabolism during germination.

Effect of germination pretreatment on the physicochemical properties and lipid concomitants of flaxseed oil.

This study investigated the effects of germination pretreatment on the physicochemical properties, lipid concomitants, and antioxidant activity of flaxseed oil in three varieties. The results indicated that the oil content of flaxseed decreased by 2.29-7.40% during the 5 days germination period. Germinated flaxseed oil showed a significantly higher acid value and lower peroxide value. The unsaturated fatty acid content was slightly increased by germination. Germination pretreatment resulted in significant increases in the α-tocopherol, stigmasterol, pigments, total phenols, and antioxidant activity. As germination time progressed to 5 days, α-tocopherol which was traditionally recognized as having the highest antioxidant activity form of vitamin E in humans increased from 3.07-6.82 mg kg-1 to 258.11-389.78 mg kg-1. Germinated oil had 1.63 to 2.05 times higher stigmasterol content than non-germinated oil. The chlorophyll and carotenoid also increased exponentially. The total phenol content of flaxseed oil increased from 64.29-75.85 mg kg-1 to 236.30-297.78 mg kg-1. Germinated flaxseed oil showed important antioxidant activity. Compared with other varieties during germination, the oil from Gansu showed a higher level of α-linolenic acid, tocopherols, and carotenoid, and a maximum increase level of tocopherols and phytosterols. The comprehensive evaluation of germination time by correlation and principal component analysis showed that when germination time exceeded 2 days, the lipid concomitants and antioxidant capacity of flaxseed oil were significantly improved.

BRD9 inhibition promotes PUMA-dependent apoptosis and augments the effect of imatinib in gastrointestinal stromal tumors.

Gastrointestinal stromal tumors (GISTs) are primarily characterized by activating mutations of tyrosine kinase or platelet-derived growth factor receptor alpha. Although the revolutionary therapeutic outcomes of imatinib are well known, the long-term benefits of imatinib are still unclear. The effects of BRD9, a recently identified subunit of noncanonical BAF complex (ncBAF) chromatin remodeling complexes, in GISTs are not clear. In the current study, we evaluated the functional role of BRD9 in GIST progression. Our findings demonstrated that the expression of BRD9 was upregulated in GIST tissues. The downregulation or inhibition of BRD9 could significantly reduce cellular proliferation, and facilitates apoptosis in GISTs. BRD9 inhibition could promote PUMA-dependent apoptosis in GISTs and enhance imatinib activity in vitro and in vivo. BRD9 inhibition synergizes with imatinib in GISTs by inducing PUMA upregulation. Mechanism study revealed that BRD9 inhibition promotes PUMA induction via the TUFT1/AKT/GSK-3ß/p65 axis. Furthermore, imatinib also upregulates PUMA by targeting AKT/GSK-3ß/p65 axis. In conclusion, our results indicated that BRD9 plays a key role in the progression of GISTs. Inhibition of BRD9 is a novel therapeutic strategy in GISTs treated alone or in combination with imatinib.

Regulatory Roles of Tumor Necrosis Factor-α-Induced Protein 8 Like-Protein 2 in Inflammation, Immunity and Cancers: A Review.

Tumor necrosis factor-alpha (TNF-α)-induced protein 8 (TNFAIP8/TIPE) family, including TNFAIP8 (TIPE), TNFAIP8 like-protein 1 (TNFAIP8L1/TIPE1), TNFAIP8 like-protein 2 (TNFAIP8L2/TIPE2), and TNFAIP8 like-protein 3 (TNFAIP8L3/TIPE3), plays a vital role in regulating inflammatory responses, immune homeostasis, and cancer development. Over the last decade, studies have shown that TIPE2 protein is differentially expressed in diverse cells and tissues. The dysregulation of TIPE2 protein can lead to dysregulation of inflammatory responses and immune homeostasis, and change the basic characteristics of cancers. In consideration of the immeasurable values of TIPE2 in diagnosis, treatment, and prognosis of various human diseases, this review will focus on the expression pattern, structure, and regulatory roles of TIPE2 in inflammation, immunity, and cancers.

Bromodomain and extraterminal domain inhibitor enhances the antitumor effect of imatinib in gastrointestinal stromal tumours.

In gastrointestinal stromal tumours (GISTs), the function of bromodomain-containing 4 (BRD4) remains underexplored. BRD4 mRNA abundance was quantified in GISTs. In the current study, we investigated the role of BRD4 in GISTs. Our results show a significant enhancement in BRD4 mRNA and a shift from very low-risk/low-risk to high-risk levels as per NCCN specifications. Overexpression of BRD4 correlated with unfavourable genotype, nongastric location, enhanced risk and decreased disease-free survival, which were predicted independently. Knockout of BRD4 in vitro suppressed KIT expression, which led to inactivation of the KIT/PI3K/AKT/mTOR pathway, impeded migration and cell growth and made the resistant GIST cells sensitive to imatinib. The expression of KIT was repressed by a BRD4 inhibitor JQ1, which also induced myristoylated-AKT-suppressible caspases 3 and 9 activities, induced LC3-II, exhibited dose-dependent therapeutic synergy with imatinib and attenuated the activation of the PI3K/AKT/mTOR pathway. In comparison with their single therapy, the combination of JQ1/imatinib more efficiently suppressed the growth of xenografts and exhibited a reduction in KIT phosphorylation, a decrease in Ki-67 and in the levels of phosphorylated PI3K/AKT/mTOR and enhanced TUNEL staining. Thus, we characterized the biological, prognostic and therapeutic implications of overexpressed BRD4 in GIST and observed that JQ1 suppresses KIT transactivation and nullifies the activation of PI3K/AKT/mTOR, providing a potential strategy for treating imatinib-resistant GIST through dual blockade of KIT and BRD4.

BRD4 promotes tumor progression and NF-κB/CCL2-dependent tumor-associated macrophage recruitment in GIST.

The most commonly occurring sarcoma of the soft tissue is gastrointestinal stromal tumor (GIST). Treatment and prevention of the disease necessitate an understanding of the molecular mechanisms involved. However, the role of BRD4 in the progression of GIST is still unclear. While it is known there are abundant infiltrating tumor-associated macrophages (TAMs) in the tumor microenvironment, the exact role of these cells has yet to be studied. This work showed an upregulation of BRD4 in GIST that was associated with GIST prognosis. Through gain and loss of function studies, it was found that BRD4 promotes GIST growth and angiogenesis in vitro and in vivo. Mechanistically, BRD4 enhances CCL2 expression by activating the NF-κB signaling pathway. Furthermore, this CCL2 upregulation causes recruitment of macrophages into the tumor leading to tumor growth. A likely mechanism for interactions in the GIST microenvironment has been outlined by this work to show the role and potential use of BRD4 as a treatment target in GIST.

Expression of miR-124 in gastric adenocarcinoma and the effect on proliferation and invasion of gastric adenocarcinoma SCG-7901 cells.

Expression of miR-124 in gastric adenocarcinoma cell line SGC-7901 and its effect on biological functions was investigated. Expression of miR-124 in cancer tissues and paracancerous tissues of gastric adenocarcinoma patients was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). RT-qPCR was used to detect the expression of miR-124 in human normal gastric epithelial cells GES-1 and gastric adenocarcinoma SGC-7901 cells. Cells in miR-124 group were transfected with miR-124 agomir, cells in NC group were transfected with agomir-negative control sequence and cells in the control group were not transfected. MTT assay was used to detect cell proliferation, and Transwell invasion assay to detect cell invasion ability, and the effect of transfected miR-124 agonist on the proliferation and invasive ability of gastric adenocarcinoma cells was evaluated. RT-qPCR results showed that miR-124 expression was significantly downregulated in gastric adenocarcinoma tumor tissues compared with paracancerous tissues. Compared with cells of normal human gastric epithelial cell line GES-1, the expression of miR-124 human gastric adenocarcinoma SGC-7901 cells was significantly downregulated. At 12 h, there was no significant difference in OD at 490 nm in the three groups (P>0.05). OD (490) in the three groups showed a gradual upward trend. After transfection, proliferation curves of the three groups showed an upward trend, proliferation rate of miR-124 group was significantly lower than that of NC and control groups (P<0.05). The number of invading cells in miR-124 group was significantly lower than that in NC group and control group, but there was no significant difference in the number of cell invasion between the NC and control groups. miR-124 can inhibit the proliferation and invasion of gastric adenocarcinoma cells. Downregulation of miR-124 expression in gastric adenocarcinoma may be closely related to the development of gastric adenocarcinoma.

Curcumin derivative L6H4 inhibits proliferation and invasion of gastric cancer cell line BGC-823.

Curcumin and its chalcone derivatives have well-known, explicit biological antitumor properties, such as instance antiproliferative and apoptotic effects via multiple molecular targets. In this study, we investigated the anticancer activity of curcumin derivative L6H4 (curcumin L6H4) on gastric cancer cells. Inhibitory effects of curcumin L6H4 on gastric cancer cells (BGC-823) were studied by the diphenyltetrazolium (MTT) assay, and cell apoptosis was detected by Annexin-V/propidium iodide (PI) staining and then analyzed by flow cytometry. A mouse xenotransplant gastric tumor model was established to detect the role of curcumin L6H4 in vivo. The apoptosis-related proteins p53, p21, Bax, and Bcl-2 in BGC-823 cells and mouse xenotransplant models treated with curcumin L6H4 were determined by Western blot analysis. Curcumin L6H4 can significantly inhibit the proliferation and induce the apoptosis of BGC-823 cells, thus enhancing the expression levels of p53, p21, Bax, and Bcl-2 noticeably in vivo and in vitro. Meanwhile, curcumin L6H4 can remarkably suppress the growth of tumor cells in animal models. These results suggest that curcumin derivative L6H4 has potent of antitumor properties in vitro or in vivo.

Eukaryotic translation initiation factor EIF3H potentiates gastric carcinoma cell proliferation.

Eukaryotic translation initiation factor 3 subunit H (EIF3H) is required for the progression of several types of cancer. However, little is known about the function of EIF3H in gastric carcinoma. To address this issue, in the present study, we investigated EIF3H genetic alterations in and expression of EIF3H in gastric cancer tissue samples using cBioPortal and Oncomine databases. Endogenous EIF3H expression was knocked down in MGC80-3 and AGS gastric cancer cell lines by lentivirus-mediated RNA interference. We confirmed the knockdown efficiency by quantitative real-time PCR and western blotting and evaluated the effects of EIF3H silencing on cell proliferation of gastric cancer with the cell viability and colony formation assays and by flow cytometry. The OncoPrint of EIF3H generated using cBioPortal indicated that EIF3H genetic alterations (mutation, deletion and amplification) were present in two gastric cancer sample sets. The Oncomine analysis revealed that EIF3H mRNA level was upregulated in gastric cancer tissues. EIF3H knockdown inhibited cell proliferation and colony formation in gastric cancer lines and led to cell cycle arrest at the G0/G1 phase, while inducing apoptosis via up- and downregulation of pro- and anti-apoptotic factors, respectively. These results indicate that EIF3H can serve as a novel therapeutic target for the clinical treatment of gastric cancer.

Effects of Scutellaria barbata polysaccharide on the proliferation, apoptosis and EMT of human colon cancer HT29 Cells.

A water-soluble polysaccharide SPS2p was isolated from the whole grass of Scutellaria barbata and SPS2p contained 53.6% carbohydrates, 38.5% uronic acid and 8.2% proteins. The molecular weight of SPS2p showed only one molecular weight distribution (2.6×104Da) and the monosaccharide composition of SPS2p showed the presence of arabinose, mannose, glucose and galactose at the ratio of 1.31:1.00:3.59:1.59. The results showed that SPS2p could improve the proliferation inhibition rate; SPS2p could also elevate apoptosis rate, apoptosis index and the levels of Bax and Bak, but lower levels of Bcl-2 and FN; SPS2p could up-regulate the expression of E-cadherin mRNA, and down-regulate the expressions of N-cadherin and vimentin mRNA, and the ratio of p-AKT/AKT in HT29 cells. These results indicate that SPS2p can inhibit the proliferation and EMT, and promote the apoptosis in HT29 cells, which may be related to the inhibition of SPS2p on the PI3K/AKT pathway.

[Advantage investigation of totally laparoscopic modified Roux-en-Y reconstruction].

OBJECTIVE: To investigate the clinical advantage of the application of modified Roux-en-Y reconstruction after totally laparoscopic total gastrectomy. METHODS: Clinical data of 36 patients who underwent totally laparoscopic total gastrectomy with Roux-en-Y reconstruction by one medical team for gastric adenocarcinoma between January 2014 and December 2014 in the Second Hospital of Jilin University were retrospectively analyzed. Patients were divided into classic Roux-en-Y group (CRY, 16 cases) and modified Roux-en-Y group (MRY, 20 cases) according to reconstructive methods. The data concerning the intraoperative and postoperative situation in two groups were compared. RESULTS: Operation was successfully completed in all the cases without conversion to laparotomy. Compared to CRY group, MRY group had shorter mean operative time [(260.9 ± 21.2) min vs. (287.9 ± 19.0) min, P=0.000], shorter mean reconstruction duration [(32.4 ± 9.2] min vs. (45.4 ± 13.2) min, P=0.001] and less intraoperative bleeding [(50.9 ± 23.5) ml vs. (67.0 ± 20.5) ml, P=0.000]. Jejunum mesentery dissection and jejunum resection were not necessary in MRY group. However, there were no significant differences in lymph nodes harvested, time to flatus, hospital stay and postoperative complications between two groups. CONCLUSIONS: As compared to classic Roux-en-Y reconstruction, the modified Roux-en-Y reconstruction can simplify the surgical procedures and achieve similar efficacy. It is feasible and safe, and worth further promotion in clinical practice.

miR-206 is an independent prognostic factor and inhibits tumor invasion and migration in colorectal cancer.

microRNAs (miRNAs) are small non-coding RNAs that play important role in a variety of biological processes, and aberrant expression of miRNA is always associated with tumor progression and invasion. However, the role of miR-206 in colorectal cancer (CRC) is still unclear. In this study, the expression of miR-206 in CRC tissue was compared to paired normal adjacent tissue, and results indicate that miRNA-206 levels are significantly lower in CRC tissue compared to control tissue. Kaplan-Meier analysis showed that patients with low miR-206 expression had significantly poorer overall survival. In addition, we found that over-expression of miR-206 can inhibit CRC cell migration and invasion. In conclusion, our results suggest that miR-206 functions as a tumor suppressor by inhibiting CRC migration and invasion. These findings suggest that miR-206 may be useful as a new potential therapeutic target for CRC.

Expression, purification and characterization of human interferon-γ in Pichia pastoris.

Human interferon-γ (hIFN-γ) is a multifunctional protein known to possess immunoregulatory, antiviral and anticancer functions. In the present study, in order to explore the biological roles of hIFN-γ and its mechanisms of action, IFN-γ was cloned and expressed in Pichia pastoris (P. pastoris) under the control of alcohol oxidase promoter 1 (AOX1). The protein was secreted by two different signal peptides, the native secretion signal peptide of hIFN-γ and the Saccharomyces cerevisiae α signal peptide. Following 96 h of methanol induction, Tricine-SDS-PAGE Coomassie staining, western blot analysis and N-terminal protein sequencing revealed that the level of recombinant hIFN-γ (rhIFN-γ) secreted by the native secretion signal was barely detectable, while the α signal peptide secreted ~300 mg/l. rhIFN-γ was purified by Vivaflow 200, SP Sepharose Fast Flow and Vivaspin 2 ml, yielding >96% of a highly purified rhIFN-γ preparation, with a specific activity of 1 x 10(7)-1.4 x 10(7) IU/mg protein as determined by an antiviral assay. The results demonstrated that the experimental procedures developed are capable of producing a large quantity of active rhIFN-γ from P. pastoris.

[Value of radical dissection with vagus nerve preservation for proximal gastric cancer].

OBJECTIVE: To study the feasibility and influence of vagus nerve preservation in radical operation for proximal gastric cancer. METHODS: Thirty-two patients with early or T2 cardia cancer from May 2007 to May 2009 were enrolled and randomized into two groups, i.e. vagus nerve preservation group(n=16) and control group(n=16). Two groups were compared with regard to operative time, anastomotic fistula, digestive discomforts, body weight, survival rate, findings on gastroscope and abdominal ultrasonography. RESULTS: There were no statistically significant differences between the two groups in operative time (2.8 vs. 2.5 h), postoperative complications rate (25.0% vs. 31.3%). No recurrence or mortality was observed after one-year follow-up. However, patients who underwent vagus nerve preservation had less postprandial discomforts(3 vs. 12 cases), bile reflux(3 vs. 10 cases), atrophic gastritis(1 vs. 9 cases), gallstones(1 vs. 8 cases), body mass index, and diarrhea(P<0.05). CONCLUSION: For patients with early gastric cancer, preservation of the vagus nerve during radical gastrectomy results in less complications and does not compromise patient survival.