Gastric Cancer Invasion Research Articles

Dextran Sulfate Inhibits Angiogenesis and Invasion of Gastric Cancer by Interfering with M2-type Macrophages Polarization.

To explore the effect of dextran sulfate (DS) on the angiogenesis, invasion, and migration of gastric cancer cells by interfering with the polarization of M2-type macrophages. The infiltration of M2-type macrophages and microvascular density in gastric cancer and paracancerous tissues were analyzed using immunohistochemistry and immunofluorescence. The effects of DS on M2-type macrophages and the angiogenesis in metastatic tumors were investigated in the nude mice intraperitoneal metastasis model using immunohistochemistry and western blot. The differentiation and polarization of macrophages, immunocytochemistry, western blot, ELISA, and transwell migration assay were used to evaluate the effect of DS on the polarization of macrophages, immunocytochemistry, western blot, ELISA, and transwell migration assay were used to evaluate the effect of DS on the polarization and recruitment capacity of macrophages. Immunocytofluorescence, tube formation assay, transwell invasion assay, wound healing assay, and western blot were used to investigate the effect of DS on the angiogenesis, invasion, and migration-promoting phenotype of M2- type macrophage in a co-culture system of macrophages and gastric cancer cells. The infiltration of M2-type macrophages and the microvascular density were highly expressed and positively correlated in the human gastric cancer tissue. DS can significantly inhibit the intraperitoneal metastases of gastric cancer in nude mice, and reduce the infiltration of M2-type macrophages and the angiogenesis in intraperitoneal metastatic tumors. Moreover, DS can prevent the polarization of M0-type macrophages to M2 type, reduce the expression of M2-type macrophage markers (CD206, CD163, IL-10, and Arg-1), down-regulate the IL-6-STAT3 pathway, and inhibit the recruitment capability of M2-type macrophages. Finally, the co-culture experiment showed that DS significantly reduced the enhancing effects of M2-type macrophages on the angiogenesis, invasion, and migration of gastric cancer cells, as well as down-regulated the related expressions of proteins (VEGF, N-cadherin, MMP-2 and Vimentin) in gastric cancer cells. DS can reduce the infiltration of M2-type macrophages and the microvascular density in intraperitoneal metastases of gastric cancer in nude mice, and inhibit the angiogenesis, invasion, and migration of gastric cancer cells by interfering with the polarization of M2-type macrophages through repression of the IL-6/STAT3 signaling pathway.

Clinicopathologic significance of human epidermal growth factor receptor 2 expression and gene amplification in gastric carcinoma.

This study is aimed at determining the human epidermal growth factor receptor-2 (HER-2) gene status in gastric cancers using immunohistochemistry (IHC) and silver DNA in situ hybridization (SISH) technique and investigating the relationship between HER-2 status and clinicopathological variables. We examined 104 gastric carcinoma cases with gastrectomy specimens. The age, sex, histopathological tumor type, tumor differentiation, tumor localization, tumor size, invasion depth, lymph node metastasis status, HER-2 IHC evaluation results, and SISH evaluation results of the cases were recorded. In the present study, the SISH-confirmed HER-2 positivity rate (SISH amplification) is 14.4%. A statistically significant relationship was found between HER-2 status and tumor invasion depth (P = 0.03). It was observed that as the depth of invasion increased, HER-2 positivity decreased. No statistically significant relationship was found between HER-2 status and sex, age, tumor size, differentiation, histopathological classification, lymph node metastasis, and tumor localization (P > 0.05). The rate of HER-2 2+ cases was found to be high. HER-2 status is negatively correlated with the depth of invasion in gastric cancers, and HER-2 positivity increases as the depth of invasion decreases. HER-2 may be early molecular evidence in oncogenesis. Immunohistochemically, HER-2 2+ cases should be confirmed by the in situ hybridization (ISH) technique.

Sympathetic nerve infiltration promotes stomach adenocarcinoma progression via norepinephrine/β2-adrenoceptor/YKL-40 signaling pathway

ObjectiveThis study aimed to address the status, role, and mechanism of sympathetic nerve infiltration in the progression of stomach adenocarcinoma (STAD). MethodsSympathetic nerve and its neurotransmitter NE, β-ARs, and associated signaling molecules in the STAD tissues and the adjacent tissues from 46 STAD patients were examined using immunostaining, HPLC, and western blotting. The effects and mechanisms of β2-AR activation on the proliferation, migration and invasion of AGS and SGC-7901 gastric cancer (GC) cell lines were examined using CCK-8, transwell, and western blotting assays. Correlations between genes and STAD survival were analyzed using bioinformatics. ResultsStriking sympathetic nerve infiltration, elevations of NGF, TrkA, GAP43, TH, S100, NE, β2-AR, YKL-40, syndecan-1, MMP9, CD206, and CD31 were observed in the STAD tissues compared to the adjacent tissues. Activation of β2-AR in the two GC cell lines significantly amplified the expressions of NGF, YKL-40, MMP9, syndecan-1, p-STAT3 and p-ERK, and increased GC cell proliferation, migration and invasion. Bioinformatic analyses revealed positive correlations of NGF, β2-AR, syndecan-1, and macrophage infiltration, respectively, with low survival of STAD, of β2-AR respectively with STAT3, ERK1/2 (MAPK1/3), YKL-40, MMP9, and syndecan-1, and of YKL-40 with MMP9. ConclusionSympathetic nerves significantly infiltrated into human STAD tissues as a result of high NGF and TrkA expressions; elevated NE led to overactivation of β2-AR-STAT3/ERK-YKL-40 signaling pathway, and finally caused cancer cell growth and invasion, M2 macrophage infiltration, angiogenesis, matrix degradation and STAD metastasis and progression.

Inflammatory Ratios as Predictors for Tumor Invasiveness, Metastasis, Resectability and Early Postoperative Evolution in Gastric Cancer.

Our study aimed to evaluate the baseline neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), and systemic immune-inflammation index (SII) in relation to invasion, metastasis, and resectability for patients with gastric cancer, respectively, as predictors of death during hospitalization or surgical complications. A retrospective cohort study was conducted on 657 gastric cancer subjects. Inflammatory biomarkers were computed. The associations with tumor stage, metastasis, optimal procedure, in-hospital mortality, and surgical complications were evaluated. Subjects who underwent curative-intent surgery presented lower median NLRs (2.9 vs. 3.79), PLRs (166.15 vs. 196.76), and SIIs (783.61 vs. 1122.25), and higher LMRs (3.34 vs. 2.9) than those who underwent palliative surgery. Significantly higher NLRs (3.3 vs. 2.64), PLRs (179.68 vs. 141.83), and SIIs (920.01 vs. 612.93) were observed for those with T3- and T4-stage cancer, in comparison with those with T1- and T2-stage cancer. Values were significantly higher in the case of metastasis for the NLR (3.96 vs. 2.93), PLR (205.22 vs. 167.17), and SII (1179 vs. 788.37) and significantly lower for the LMR (2.74 vs. 3.35). After the intervention, the NLR, PLR, and SII values were higher (p < 0.01) for patients with surgical complications, and the NLR and SII values were higher for those who died during hospitalization. Higher NLRs, PLRs, SIIs, and lower LMRs were associated with a more aggressive tumor; during early follow-up, these were related to post-operative complications and death during hospitalization.

ENKUR recruits FBXW7 to ubiquitinate and degrade MYH9 and further suppress MYH9‐induced deubiquitination of β‐catenin to block gastric cancer metastasis

ENKUR was shown as a suppressor in some tumors. However, the biological role of ENKUR on gastric cancer (GC) and its related molecular mechanisms is not clear. Here, we first observed that ENKUR significantly inhibited cell migration, invasion, and metastasis in GC. The molecular basis showed β-catenin-mediated epithelial-mesenchymal transition (EMT) signaling was inactivated in ENKUR-overexpressing GC cells. In addition, ENKUR knockdown markedly restored cell migration and invasion. Subsequently, ENKUR bound to MYH9 and decreased its protein expression by recruiting E3 ubiquitin ligase FBXW7 to form an ubiquitinated degradation complex. The downregulated MYH9 protein weakened the recruitment of the deubiquitinase USP2 and thus promoted the degradation of β-catenin protein, which finally suppressed EMT signaling. Finally, the oncogenic transcription factor c-Jun bound to ENKUR promoter and reduced its expression in GC. In clinical samples, decreased ENKUR expression promoted the unfavorable prognosis of GC. Our data proved the vital role of ENKUR on suppressing cell migration, invasion, and metastasis and demonstrated its potential as a therapeutic target for GC.

ZNF521/EBF1 axis regulates AKR1B1 to promote the proliferation, migration, and invasion of gastric cancer cells.

Although the incidence and death rates of gastric cancer (GC) are decreasing, approximately one million new cases and 800,000 GC-related deaths were reported worldwide in 2018. Currently, the oncogenesis of GC remains unclear, and the demand for novel treatment options are unmet. Here, we explored the role of aldo-keto reductase family 1 member B (AKR1B1) in the progression of GC. The proliferation, migration, and invasion of GC cells were evaluated by CCK-8 assay, wound healing assay, and transwell assay, respectively. The interaction between EBF transcription factor 1 (EBF1) and the promoter region of AKR1B1 was determined by luciferase reporter assay and chromatin immunoprecipitation (ChIP). Gene expression levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting assay. The expression of AKR1B1 was elevated in GC cells, resulting in increased cell proliferation, migration, and invasion. Meanwhile, EBF1 was a negative regulator of AKR1B1; its overexpression suppressed AKR1B1 expression and GC progression. Furthermore, knockdown of ZNF521 induced EBF1 expression, thus suppressing AKR1B1 expression and resulting in attenuated GC growth and invasiveness. Notably, knockdown of ZNF521 attenuated GC progression and was rescued by overexpression of AKR1B1. Our current study revealed a novel ZNF521/EBF1/AKR1B1 axis in GC and elaborated its important role in promoting GC progression, providing potential therapeutic targets for anti-GC treatments.

SUMOylation of TUFT1 is essential for gastric cancer progression through AKT/mTOR signaling pathway activation.

Tuftelin (TUFT1) is highly expressed in various tumor types and promotes tumor growth and metastasis by activating AKT and other core signaling pathways. However, the effects of post-translational modifications of TUFT1 on its oncogenic function remain unexplored. In this study, we found that TUFT1 was SUMOylated at K79. SUMOylation deficiency significantly impaired the ability of TUFT1 to promote the proliferation, migration, and invasion of gastric cancer (GC) cells by blocking AKT/mTOR signaling pathway activation. SUMOylation of TUFT1 is mediated by the E3 SUMO ligase tripartite motif-containing protein 27 (TRIM27), and these two proteins regulate the malignant behavior of GC cells and AKT activation in the same pathway. TUFT1 binds to TRIM27 through its N-terminus, and decreased binding affinity of TUFT1 to TRIM27 significantly impairs its oncogenic effect. In addition, data collected from GC clinical samples indicated that the combined detection of TUFT1 and TRIM27 expression reflected tumor malignancy and patient survival with higher precision. In addition, we proved that SUMOylated TUFT1 is not only an upstream signal for AKT activation but also directly activates mTOR by forming a complex with Rab GTPase activating protein 1, which further inhibits Rab GTPases and promotes the perinuclear accumulation of mTORC1. Altogether, these data indicate that SUMOylated TUFT1 is the active form that affects GC progression through the AKT/mTOR signaling pathway and might be a promising therapeutic target or biomarker for GC progression.

MiRNA-223-3p regulates ECT2 to promote proliferation, invasion, and metastasis of gastric cancer through the Wnt/β-catenin signaling pathway.

Expression of the guanine nucleotide exchange factor epithelial cell transforming 2 (ECT2) is elevated in gastric cancer (GC) but its biological function in GC is poorly understood. MicroRNAs (miRNAs) have great potential as therapeutic targets for GC through their ability to modulate gene expression. In the present study, we sought to identify potential miRNA-mRNA-protein regulatory pathways that might control ECT2 expression and function in GC. ECT2 expression was examined in clinical GC specimens by immunohistochemical staining, and protein levels were correlated with clinicopathological features and prognosis. TargetScan was used to identify potential ECT2 mRNA-complementary miRNAs, and the roles of ECT2 and miRNA-223-3p (miR-223-3p) in GC cell biology and signaling pathway activation were examined by targeted knockdown (KD) or overexpression (OE) of ECT2 and miR-223-3p in GC cell lines. A murine GC xenograft model was developed to explore the impact of ECT2 OE on tumor growth in vivo. ECT2 expression was significantly elevated in GC specimens compared with normal gastric tissues and the level correlated positively with depth of invasion, ulceration, vascular tumor thrombus, neural invasion, and lymph node metastasis (p < 0.05). ECT2 was an independent prognostic factor for overall survival of GC patients (high ECT2 expression v.s. low ECT2 expression: χ2 = 29.831, p < 0.001). ECT2 KD or miR-223-3p OE markedly suppressed the proliferation, migration, and invasion of GC cells in vitro, whereas ECT2 OE had the opposite effects. ECT2 OE also promoted the growth of GC tumors in vivo. Tumor expression of Wnt2, β-catenin, and several downstream target proteins in GC cells were decreased by ECT2 KD or miR-223-3p OE but increased by ECT2 OE. miR-223-3p regulates ECT2 expression to promote tumorigenic behavior of GC via activation of the Wnt/β-catenin signaling pathway, suggesting that ECT2 and miR-223-3p as potential therapeutic targets for GC.

Knockdown of mediator complex subunit 27 suppresses gastric cancer cell metastasis and angiogenesis via Wnt/β-catenin pathway

Mediator complex (MED) contains 28 subunits, functions as a transcription machinery through interaction with RNA polymerase II and modulates gene expression involved in cell survival and growth. MED27, as an oncogene, stimulates malignant behavior of various tumors. Role of MED27 in gastric cancer was assessed in this study. Firstly, bioinformatics analysis predicted that MED27 was elevated in gastric cancer. Gastric cancer cells also showed higher expression of MED27 than normal gastric epithelial cells. Secondly, functional assays revealed that silencing of MED27 decreased cell viability, and reduced proliferation of gastric cancer. Cell invasion and migration of gastric cancer were also inhibited by loss of MED27. Moreover, knockdown of MED27 inhibited angiogenesis of gastric cancer. Thirdly, nuclear protein of β-catenin in gastric cancer was reduced by silencing of MED27. Lastly, in vivo tumor growth of gastric cancer was suppressed by interference of MED27. In conclusion, MED27 functioned as an oncogene in gastric cancer through promoting cell metastasis and angiogenesis.

Knockdown of LINC01279 Suppresses Gastric Cancer Proliferation and Migration by Inhibiting PI3K/Akt/mTOR Signaling Pathway.

To explore the functional and molecular mechanism of long noncoding RNA LINC01279 in gastric cancer (GC). The LINC01279 expression in GC and tissues of para-carcinoma was detected by qPCR (real-time fluorescent quantitative PCR), and the association between the LINC01279 expression and clinicopathological features of patients with GC was investigated. The colony formation, CCK-8, transwell assays, and cell cycle detection kit were used for detection of the effect of LINC01279 on GC cell proliferation, cell cycle, colony formation, and invasion. The effect of LINC01279 on PI3K/AKT/mTOR in the GC signaling pathway was identified by the Western blotting technique. The effect of LINC01279 on GC cell proliferation in vivo was evaluated by subcutaneous xenograft tumors in the nude mice. The results of qPCR displayed the expression of LINC01279 was higher in tissues of GC patients. Furthermore, the tumor size, TNM stage, and metastasis of lymph nodes were also closely related to LINC01279 expression. The experiments on cell function showed that the LINC01279 knockdown significantly inhibited the colony formation, invasion, and proliferation of GC cells and induced the cell cycle arrest in G0 and G1 phases. The Western blotting technique also showed that LINC01279 knockdown significantly inhibited the phosphorylation of PI3K, Akt, and mTOR in GC cells. Furthermore, in vivo experiments displayed that the LINC01279 knockdown significantly inhibited the GC growth. Knockdown of LINC01279 plays a significant role in inhibiting the PI3K/AKT/mTOR signaling pathway which affects the GC invasion and proliferation. The LINC01279 expression can be utilized as a biomarker for the prediction of the GC prognosis.

Upregulated Solute Carrier SLC39A1 Promotes Gastric Cancer Proliferation and Indicates Unfavorable Prognosis.

Solute carrier 39A1 (SLC39A1) is an indirect zinc transporter which showed diverse tumor-related functions in different malignancies. Here, we aimed to investigate its expression and role in gastric adenocarcinoma. A retrospective gastric adenocarcinoma cohort (n = 154) was collected from our hospital to test their tissue expression of SLC39A1 through immunohistochemical staining method. After SLC39A1 overexpression or knockdown, proliferation and invasion assays were conducted for proliferation and invasion estimation, respectively. Xenograft in nude mice was used as the in vivo strategy to validate in vitro findings. Compared with adjacent stomach tissues, gastric adenocarcinoma tissues showed significantly higher SLC39A1 on both mRNA and protein levels. Higher SLC39A1 was observed in patients with larger tumor size (P=0.003) and advanced tumor stages (P < 0.001). Univariate (P=0.001) and multivariate analyses (P=0.035) confirmed the independent prognostic significance of SLC39A1 on gastric adenocarcinoma outcomes. The median survival time was 22.0 months in patients with high-SLC39A1 expression, while up to 57.0 months in those with low-SLC39A1 (P=0.001). In vitro and in vivo assays demonstrated that overexpressing SLC39A1 could promote gastric cancer growth and invasion, while silencing SLC39A1 led to opposite effects. Aberrant high-SLC39A1 expression can serve as an independent unfavorable prognostic factor for gastric adenocarcinoma. High SLC39A1 is critical for a more aggressive tumor phenotype by promoting cell proliferation and invasion. Therefore, targeting SLC39A1 may provide novel therapeutic insights.

Low FNDC5/Irisin expression is associated with aggressive phenotypes in gastric cancer.

Background: FNDC5 belongs to the family of proteins called fibronectin type III domain-containing which carry out a variety of functions. The expression of FNDC5 is associated with the occurrence and development of tumors. However, the role of FNDC5 in gastric cancer remains relatively unknown. Methods: In the research, the expression of FNDC5 and its value for the prognosis of gastric cancer patients were observed with the TCGA database and GEO datasets of gastric cancer patients. The role of FNDC5 in the regulation of gastric cancer cells proliferation, invasion, and migration was determined. WGCNA and Enrichment analysis was performed on genes co-expressed with FNDC5 to identify potential FNDC5-related signaling pathways. Meanwhile, the LASSO Cox regression analysis based on FNDC5-related genes develops a risk score to predict the survival of gastric cancer patients. Results: The expression of FNDC5 was decreased in gastric cancer tissues compared to normal gastric tissues. However, survival analysis indicated that lower FNDC5 mRNA levels were associated with better overall survival and disease-free survival in gastric cancer patients. Meanwhile, a significant negative correlation was found between FNDC5 and the abundance of CD4+ memory T cells in gastric cancer. In vitro overexpression of FNDC5 inhibits the migration and invasion of gastric cancer cells, without affecting proliferation. Finally, A two-gene risk score module based on FNDC5 co-expressed gene was built to predict the overall clinical ending of patients. Conclusion: FNDC5 is low expressed in gastric cancer and low FNDC5 predicts a better prognosis. The better prognosis of low FNDC5 expression may be attributed to the increased number of CD4+ memory activated T-cell infiltration in tumors, but the exact mechanism of the effect needs to be further explored. Overexpressing FNDC5 inhibits the invasion and migration of gastric cancer but does not affect proliferation. At last, we constructed a clinical risk score model composed of two FNDC5-related genes, and this model may help lay the foundation for further in-depth research on the individualized treatment of gastric cancer patients.

CircPCSK5 is highly expressed in gastric cancer and promotes cancer cell proliferation, invasion and epithelial-mesenchymal transition

To investigate the expression of circPCSK5 in gastric cancer (GC) and its role in regulation of the proliferation, invasion and epithelial-mesenchymal transition (EMT) of GC cells. High-throughput sequencing was performed in 3 pairs of GC and adjacent gastric mucosa tissues to obtain the differential expression profile of circRNA. The expression of circPCSK5 was detected in 62 patients undergoing radical surgery for GC using RT-qPCR, and the correlation between circPCSK5 expression level and clinicopathological data of the patients was analyzed. The overall survival and disease-free survival of the patients were assessed with Kaplan-Meier survival analysis, and the independent risk factors affecting the patients' prognosis were analyzed using Cox proportional hazards regression model. The stability and subcellular localization of circPCSK5 were assessed using RNase R and actinomycin D assays, fluorescence in situ hybridization and nucleocytoplasmic separation assay. CCK-8 assay, EdU assay and Transwell assay were employed to examine the changes in proliferation, migration and invasion of GC cells with circPCSK5 knockdown or overexpression; Western blotting and RT-qPCR assays were used to detect the expression levels of EMT markers in the transfected cells. The expression of circPCSK5 was significantly upregulated in GC tissues and cells (P < 0.001, P < 0.01). The expression level of circPCSK5 was positively correlated with tumor size, vascular invasion, lymph node metastasis and AJCC stage of GC (P < 0.05). The overall survival and disease-free survival were significantly lower in GC patients with high circPCSK5 expression than in those with low circPCSK5 expression (P < 0.001). High circPCSK5 expression was an independent risk factor for a poor prognosis of GC patients (P < 0.05). Knockdown of circPCSK5 significantly inhibited the proliferation, migration and invasion of HGC27 cells (P < 0.01), increased the expressions of E-cadherin, and decreased the expression of N-cadherin and vimentin (P < 0.01). CircPCSK5 overexpression promoted the proliferation, migration and invasion of MKN45 cells (P < 0.01), reduced E-cadherin expression and increased N-cadherin and vimentin expressions (P < 0.01). CircPCSK5 is highly expressed in GC and promotes the proliferation, invasion and EMT of GC cells, suggesting its potential as a prognostic biomarker and therapeutic target for GC.

ZNF521 Is Correlated with Tumor Immune Cell Infiltration and Act as a Valuable Prognostic Biomarker in Gastric Cancer.

Aim To explore the correlations between the expression of zinc finger protein 521 (ZNF521) with immune invasion and prognosis of gastric cancer. Methods Expression of ZNF521 was examined by immunohistochemistry in gastric cancer cases. Kaplan–Meier plotter was used to determine the relationships between ZNF521 and prognosis. TIMER and GEPIA were used to analyze the correlation between ZNF521 expression and gene markers of immune cell infiltration. Results The expression of ZNF521 was up-regulated in gastric cancer samples. Kaplan–Meier analysis indicated that higher expression of ZNF521 was associated with poor prognosis. The expression of ZNF521 was correlated with infiltrating levels of CD4+ T and CD8+ T cells, macrophages, neutrophils, and dendritic cells in gastric cancer, which also correlated with diverse immune marker sets. Conclusions ZNF521 is correlated significantly with immune cell infiltration and is a valuable biomarker for prognosis in gastric cancer.

Spectral CT for preoperative prediction of lymphovascular invasion in resectable gastric cancer: With external prospective validation.

ObjectivesTo develop and externally validate a spectral CT based nomogram for the preoperative prediction of LVI in patients with resectable GC.MethodsThe two centered study contained a retrospective primary dataset of 224 pathologically confirmed gastric adenocarcinomas (161 males, 63 females; mean age: 60.57 ± 10.81 years, range: 20-86 years) and an external prospective validation dataset from the second hospital (77 males and 35 females; mean age, 61.05 ± 10.51 years, range, 31 to 86 years). Triple-phase enhanced CT scans with gemstone spectral imaging mode were performed within one week before surgery. The clinicopathological characteristics were collected, the iodine concentration (IC) of the primary tumours at arterial phase (AP), venous phase (VP), and delayed phase (DP) were measured and then normalized to aorta (nICs). Univariable analysis was used to compare the differences of clinicopathological and IC values between LVI positive and negative groups. Independent predictors for LVI were screened by multivariable logistic regression analysis in primary dataset and used to develop a nomogram, and its performance was evaluated by using ROC analysis and tested in validation dataset. Its clinical use was evaluated by decision curve analysis (DCA).ResultsTumor thickness, Borrmann classification, CT reported lymph node (LN) status and nICDP were independent predictors for LVI, and the nomogram based on these indicators was significantly associated with LVI (P<0.001). It yielded an AUC of 0.825 (95% confidence interval [95% CI], 0.769-0.872) and 0.802 (95% CI, 0.716-0.871) in primary and validation datasets (all P<0.05), with promising clinical utility by DCA.ConclusionThis study presented a dual energy CT quantification based nomogram, which enables preferable preoperative individualized prediction of LVI in patients with GC.

MiR-28-5p's Targeting of GAGE12I Inhibits Proliferation, Migration, and Invasion of Gastric Cancer in Vitro.

GAGE12I is a tumor metastasis-promoting factor, which can induce gastric cancer cells to invade and migrate. We investigated the effect of miR-28-5p targeting GAGE12I on proliferation, invasion, and migration of human gastric cancer cell lines SGC-7901, AGS, and MGC-803. The expression levels of miR-28-5p and GAGE12I were detected by real-time PCR and western blot, respectively. Cell proliferation, migration, and invasion were measured by MTT and Transwell chamber. The interaction between miR-28-5p and GAGE12I was investigated by bioinformatics analysis and luciferase assay. Results showed that the expression of miR-28-5p in human gastric cancer cell lines was lower than that in normal gastric epithelial cells (P < 0.05). Overexpression of miR-28-5p suppressed cell proliferation, invasion, and migration (P < 0.05). GAGE12I was confirmed as a target of miR-28-5p. Cell proliferation, invasion, and migration were decreased in cells transfected with shGAGE12I compared with those of the scrambled group (P < 0.05). Collectively, miR-28-5p negatively regulated GAGE12I and reduced the proliferation, invasion, and migration of gastric cancer cells.

PRSS2 overexpression relates to poor prognosis and promotes proliferation, migration and invasion in gastric cancer

Serine protease 2 (PRSS2) plays a pivotal role in tum or pathogenesis as a serine protease. In this paper, we investigated the expression and role of PRSS2 in gastric cancer (GC). Quantitative real-time polymerase chain reaction (qRT-PCR), western blotting and immunohistochemistry (IHC) were performed to detect PRSS2 expression in GC tissues. The correlations between PRSS2 and clinicopathological variables and prognosis were analyzed. The effects of PRSS2 on gastric cancer in vitro and in vivo were detected by Cell Counting Kit-8 (CCK-8), colony formation assay, migration and invasion test, and nude mouse experiment. We observed that the PRSS2 mRNA and protein levels were upregulated in GC tissues, and PRSS2 expression was associated with GC N stage. Patients with high PRSS2 expression have a poor prognosis. Finally, the regulatory effect of PRSS2 on the biological behaviors of GC cells was assessed in GC cells transfected with lentiviral interference fragments of PRSS2. The results suggested that PRSS2 knockdown decreased the proliferation, migration and invasion of GC cells and downregulated MMP-9 expression. In summary, PRSS2 overexpression is associated with poor postoperative prognosis in GC patients and can be used as a potential biomarker to assess the prognosis of GC.

Downregulation of long noncoding RNA HCP5/miR-216a-5p/ZEB1 axis inhibits the malignant biological function of laryngeal squamous cell carcinoma cells.

Previous studies find that long noncoding RNA human leukocyte antigen complex P5 (HCP5) is regarded as an oncogene via accelerating cancer cell growth, invasion, metastasis, vascularization, and drug resistance in renal cell carcinoma, gastric cancer, and colorectal cancer. Nevertheless, the effect and regulatory mechanism of HCP5 in laryngeal squamous cell carcinoma (LSCC) remains unknown. In this study, HCP5 expression levels were confirmed to be prominently raised in LSCC cell lines. HCP5 knockdown reduced cell proliferation and migration and invasive ability of LSCC cell lines. Furthermore, miR-216a-5p was confirmed to sponge HCP5, and its expression was prominently downregulated in LSCC cell lines and upregulated in HCP5-silenced LSCC cell lines. miR-216a-5p overexpression downregulated the cell proliferation and migration and invasive ability of LSCC cells. Additionally, the protein level of zinc finger E-box binding homeobox 1 (ZEB1), one target gene of miR-216a-5p, was highly expressed in LSCC cell lines, and its expression level was downregulated by HCP5 knockdown and miR-216a-5p overexpression. An miR-216a-5p inhibitor reversed the effect of HCP5 knockdown on the proliferation and migration and invasive ability of LSCC cells. In conclusion, knocking down HCP5 may be a strategy to suppress the malignant biological function via regulating miR-216a-5p/ZEB1. Therefore, HCP5 may become a prospective therapeutic target for LSCC.

Galectin‑1 binds GRP78 to promote the proliferation and metastasis of gastric cancer.

The present study aimed to investigate the potential molecular mechanisms by which galectin-1 (Gal-1) and glucose-regulated protein 78 (GRP78) influence the development of malignant gastric cancer (GC). Immunohistochemistry and western blotting were used to map the expression and location of the Gal-1 gene in the 80 paraffin-embedded GC samples, 16 fresh samples and surrounding tissues. Gal-1 was overexpressed and knocked down using lentiviral vectors in the human GC cell lines HGC-27 and AGS. Through the use of the Cell Counting Kit-8 assay, clone formation assay, wound healing assay, invasion assay and tumor xenograft, the possible biological roles of Gal-1 were further evaluated. The downstream interacting proteins were predicted by the BioGRID database, and GRP78 was chosen for further investigation. Immunofluorescence labeling and Co-IP were used to confirm the connection. The statistical tests utilized were the two-tailed paired Student's t-test, χ2 test, Kaplan-Meier and Cox regression analysis, and Spearman's rank correlation coefficients. In GC, Gal-1 is extensively expressed and has the potential to interact with GRP78. Poor prognosis is linked to high levels of GRP78 and Gal-1 expression in patients with GC. According to the functional study, Gal-1 knockdown prevented cells from thriving and pushed Gal-1 expression, which aided in the proliferation, migration and invasion of GC. Gal-1 overexpression additionally aided the development of subcutaneous xenograft tumors. The mechanistic investigation proved that GRP78 and Gal-1 interacted, accelerating the course of GC. Gal-1 silencing had an inhibitory effect on the proliferation of HGC-27 cells that was removed by ectopic GRP78 expression, whereas the stimulating effects of Gal-1 overexpression in AGS cells were inhibited by GRP78 knockdown. In conclusion, Gal-1 interacts with GRP78 to facilitate the advancement of GC. The Gal-1/GRP78 axis is supported by the functional data of the present study as a possible GC treatment target.

MicroRNA-34c-5p exhibits anticancer properties in gastric cancer by targeting MAP2K1 to inhibit cell proliferation, migration, and invasion.

Purpose Gastric cancer(GC)is one of the deadliest digestive tract tumors worldwide，existing studies suggest that dysregulated expression of microRNAs (miRNAs) plays an important role in the pathogenesis and progression of GC. This study aimed to investigate the expression, biological function, and downstream mechanism of miR-34c-5p in GC, provide new targets for gastric cancer diagnosis and treatment. Methods The expression of miR-34c-5p in GC tissues and cell lines was examined by RT-qPCR. Cell wound healing, transwell and cell cloning assays were used to detect the effect of miR-34c-5p on the migration and invasion abilities, respectively, of GC cells. Western blot was performed to detect the expression of related proteins. Bioinformatics analysis was used to predict the binding of MAP2K1 to miR-34c-5p and the targeting relationship was confirmed by dual luciferase reporter assay. Results The expression level of miR-34c-5p was significantly decreased in GC tissues and cell lines. miR-34c-5p overexpression inhibited migration, invasion, and colony formation of gastric cancer cells, the related protein E-cadherin expression was significantly increased and N-cadherin, vimentin, and PCNA expression were significantly decreased, while miR-34c-5p knockdown exerted the opposite effects. In addition, the targeting relationship between miR-34c-5p and MAP2K1 was predicted and confirmed, and further confirmed by rescue experiments that MAP2K1 alleviated the inhibitory effect of miR-34c-5p in GC. Conclusion MiR-34c-5p is lowly expressed in GC, and it can target MAP2K1 to exert inhibitory effects on GC proliferation, invasion, and migration. These findings provide a promising biomarker and a potential therapeutic target for gastric cancer.