Arginase 2 Promotes Colorectal Cancer Metastasis via PI3K/AKT Pathway Activation and Regulates Tumor Immune Infiltration

BackgroundColorectal cancer (CRC) is one of the most common malignant tumors of digestive system. As ubiquitin E3 ligases, Deltex (DTX) family proteins have been reported to be associated with multiple cancers. However, the role of Deltex-3 (DTX3) in CRC cells remains unclear. The study aimed to investigate the role of DTX3 in CRC.MethodsGene Expression Profiling Interactive Analysis (GEPIA) and Human Protein Atlas (HPA) were used to analyze DTX3 messenger RNA (mRNA) and protein expression in CRC and para-cancerous normal (PN) tissues, HPA was used to analyze the DTX3 expression and the survival prognosis of CRC patients. Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting (WB) and immunohistochemistry (IHC) were used to detect DTX3 mRNA and protein expression, in 80 CRC and PN tissues, and the relationships between DTX3 expression and clinicopathological characteristics of CRC patients were analyzed. DTX3 mRNA in CRC cells was knockdown or overexpressed using gene-editing tools and transfection efficiency was detected. Scratch and Transwell invasion assays were used to detect the effects of altered DTX3 mRNA expression on the migration and invasion ability of CRC cells, and the expression of epithelial-mesenchymal transition (EMT), AKT and p-AKT-related proteins were detected via WB.ResultsGEPIA data analysis showed that the expression level of DTX3 mRNA in CRC tissues was significantly lower than PN tissues. HPA analysis showed positive DTX3 protein staining in PN tissues, and the greater overall survival in CRC patients with positive DTX3 staining (P=0.007). qRT-PCR and WB revealed the mRNA and protein expression levels of DTX3 in CRC tissues were significantly lower than PN tissues (t=0.851, P<0.001; t=0.914, P<0.001), and IHC revealed the DTX3 positive staining in CRC tissues was lower than PN tissues (P<0.001). In addition, the expression of DTX3 was correlated to the depth of tumor invasion, degree of lymph node metastasis and tumor-node-metastasis (TNM) stage of CRC patients. DTX3 mRNA and protein expression in the short hairpin RNA (DTX3-shRNA) group was significantly decreased; in contrast the pcDNA3.1-DTX3 group. Scratch and Transwell experiments revealed the migration and invasion ability of CRC cells was significantly greater in the DTX3-shRNA group than the control and knockdown empty group (neg-shRNA) group, whereas the pcDNA3.1-DTX3 group was significantly lower than the control and pcDNA3.1 group. Compared with the control and pcDNA3.1 group, the average expression levels of Vimentin and p-AKT proteins in the pcDNA3.1-DTX3 group were significantly lower, whereas the E-cadherin protein level was significantly greater. In contrast, this trend was reversed when DTX3 mRNA was knockdown in CRC cells, and there was no significant difference in the expression levels of these proteins between the control and neg-shRNA or pcDNA3.1 group.ConclusionsDTX3 inhibits CRC migration, invasion and EMT through the AKT signaling pathway, and DTX3 could be used to a novel biomarker inhibit CRC progression.

BackgroundColorectal cancer (CRC) is one of the most lethal malignancies. Tripartite Motif Containing 14 (TRIM14) is a member of TRIM family proteins, which are involved in the pathogenesis of various cancers. This study aimed to investigate TRIM14 expression in CRC tissues, and its effects on the migration and invasion of CRC cell lines.MethodsTRIM14 mRNA expression was detected by real-time PCR analysis. Cell migration and invasion were measured by Transwell assays. Protein expression was assessed by western blot analysis.ResultsThe expression of TRIM14 was significantly higher in CRC tissues than in matched non-cancerous tissues. TRIM14 knockdown by specific short hairpin RNA (shRNA) attenuated CRC cell migration and invasion, whereas TRIM14 overexpression caused reverse effect. Moreover, TRIM14 positively regulated the protein levels of sphingosine kinase 1 (SPHK1) and phosphorylated STAT3 (p-STAT3), as well as the mRNA and protein expression of matrix metalloproteinase 2, MMP9 and vascular endothelial growth factor, which are transcriptional targets of the STAT3 signaling pathway. Importantly, the blockage of the SPHK1/STAT3 signaling pathway by SKI-II or AG490 could reverse the TRIM14-promoted CRC cell migration and invasion.ConclusionsOur results reveal a critical role for TRIM14 in promoting migration and invasion of CRC cells, and suggest TRIM14 may serve as a potential molecular target to prevent CRC metastasis.

BackgroundColorectal cancer (CRC) is regarded as one of the most common malignancies in the world. MiR-1-3p was reported to be a tumor suppressor in CRC. However, the mechanisms have not been fully elucidated.MethodsTo identify CRC-associated miRNA, microarray data set GSE30454 was downloaded from the Gene Expression Omnibus database (GEO), and miR-1-3p was screened out as a candidate. The expression of miR-1-3p was detected using quantitative real-time polymerase chain reaction (qRT-PCR) in CRC cell lines and tissues. CCK-8 assay and transwell invasion assay were performed to determine CRC cell line proliferation and invasion, respectively. The levels of YWHAZ and EMT-associated proteins were detected using western blotting.ResultsBioinformatic analysis showed that miR-1-3p was downregulated in CRC tissues, which is verified by our experimental validation. The overexpression of miR-1-3p significantly suppressed CRC cell proliferation and invasion. Further studies showed that YWHAZ was a direct target of miR-1-3p and mediated epithelial–mesenchymal transition (EMT) modulated by miR-1-3p.ConclusionOur results demonstrated that miR-1-3p suppresses colorectal cancer cell proliferation and metastasis through regulating YWHAZ-mediated EMT, which may support a novel therapeutic strategy for CRC patients.

Super-enhancer-associated long noncoding RNAs (SE-lncRNAs) play crucial roles in CRC pathogenesis. RP11-803D5.4 and AC005592.2 were identified as SE-lncRNAs of interest via microarray analysis, and our study aimed to evaluate their clinical value in CRC diagnosis and prognosis assessment. Fluorescence quantitative real-time PCR (qRT-PCR) was used to measure the expression of RP11-803D5.4 and AC005592.2 in the tissues and serum of CRC patients. Receiver operating characteristic (ROC) curves were generated to determine the predictive value of the two SE-lncRNAs. Functional assays were applied to assess the ability of RP11-803D5.4 to promote the proliferation, migration, and invasion of CRC cells. The two SE-lncRNAs were significantly upregulated in CRC tissue and serum samples vs. corresponding controls. ROC curve analysis indicated that RP11-803D5.4 (AUC=0.842) and AC005592.2 (AUC=0.811) had a high diagnostic performance for CRC. The combination of RP11-803D5.4, AC005592.2, and CEA had an AUC of 0.946 and distinguished CRC patients and healthy controls better than SE-lncRNA alone. The serum levels of RP11-803D5.4 and AC005592.2 were strongly correlated with their tissue expression levels. The expression levels of the two SE-lncRNAs were significantly lower in postoperative samples than in preoperative samples. Furthermore, similar to the findings of previous studies on AC005592.2, high RP11-803D5.4 expression promoted the proliferation, invasion, and migration of CRC cells. The findings suggested that RP11-803D5.4 and AC005592.2 are upregulated in CRCact and are crucial promoters of CRC progression. They also suggested that they might serve as noninvasive biomarkers for diagnosing CRC.

BackgroundColorectal cancer (CRC) is an aggressive tumor of the gastrointestinal tract, which is a major public health concern worldwide. Despite numerous studies, the precise mechanism of metastasis behind its progression remains elusive. As a member of the containing olfactomedin domains protein family, olfactomedin 2 (OLFM2) may play a role in tumor metastasis. It is highly expressed in colorectal cancer, and its role in the metastasis of CRC is still unclear. As such, this study seeks to explore the function of OLFM2 on CRC metastasis and its potential mechanisms.MethodsReal-time fluorescence quantitative PCR and western blotting were used to study the expression of OLFM2 in human CRC and adjacent normal tissues. Knockdown and overexpression OLFM2 cell lines were constructed using siRNA and overexpression plasmids to explore the role of OLFM2 in the migration and invasion of CRC through transwell, and wound healing experiments. Finally, the expression of epithelial-mesenchymal transition (EMT) -related proteins and TGF-β/Smad signaling pathway-related proteins was investigated using western blotting.ResultsIn this study, we observed an elevation of OLFM2 expression levels in CRC tissues. To investigate the function of OLFM2, we overexpressed and knocked down OLFM2. We discovered that OLFM2 knockdown inhibited migration and invasion of colon cancer cells. Furthermore, E-cadherin expression increased while N-cadherin and Vimentin expression were opposite. It is no surprise that overexpressing OLFM2 had the opposite effects. We also identified that OLFM2 knockdown resulted in reduced TGF-βR1 and downstream molecules p-Smad2 and p-Smad3, which are related to the TGF-β / Smad pathway. In contrast, overexpressing OLFM2 significantly boosted their expression levels.ConclusionThe protein OLFM2 has been identified as a crucial determinant in the progression of CRC. Its mechanism of action involves the facilitation of EMT through the TGF-β/Smad signaling pathway. Given its pivotal role in CRC, OLFM2 has emerged as a promising diagnostic and therapeutic target for the disease. These results indicate the potential of OLFM2 as a valuable biomarker for CRC diagnosis and treatment and highlight the need for further research exploring its clinical significance.

MiR-410-3p activates the NF-κB pathway by targeting ZCCHC10 to promote migration, invasion and EMT of colorectal cancer

PurposeLong non-coding RNA plasmacytoma variant translocation 1 (PVT1) has been revealed to involve in the progression of CRC. However, the precise mechanisms of PVT1 in action remain unclear.MethodsThe expression of PVT1, microRNA-106b-5p (miR-106b-5p) and four jointed box 1 (FJX1) was measured using quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot, respectively. Cell proliferation was investigated by 3-(4,5)-dimethylthiahiazo (−z-y1)-3,5-di-phenytetrazoliumromide assay. Transwell assay was used to determine cell migration and invasion. The correlation between miR-106b-5p and PVT1 or FJX1 was confirmed using luciferase reporter assay. The effects of PVT1 in vivo were assessed using mice xenograft model.ResultsPVT1 was up-regulated in CRC tissues and cell lines, especially in CRC tissues with high-grade, and highly expressed PVT1 predicted worse prognosis. Functional experiments demonstrated that PVT1 deletion inhibited CRC cell proliferation, migration and invasion in vitro and suppressed tumor growth in vivo. MiR-106b-5p was confirmed to be a target of PVT1, and inhibition of miR-106b-5p reversed the inhibitory effects of PVT1 knockdown on CRC cell malignant phenotypes. In addition, we found miR-106b-5p directly targeted FJX1, and miR-106b-5p-mediated inhibition on CRC cell proliferation, migration and invasion was attenuated by FJX1 up-regulation. Importantly, it was also proved that PVT1 could indirectly regulate FJX1 expression via targeting miR-106b-5p.ConclusionKnockdown of PVT1 impaired cell proliferation, migration and invasion in CRCs via regulating miR-106b-5p/FJX1 axis, which provided a novel insight into the development of therapeutic strategies for CRC patients.

BackgroundColorectal cancer (CRC) is the third most common cancer worldwide and the second leading cause of cancer-related death. This research focuses on investigating the impact and underlying molecular mechanisms of protocadherin gamma subfamily A, 9 (PCDHGA9) on the invasion and metastasis of CRC, aiming to identify more precise molecular markers for the diagnosis and prognosis of CRC.MethodsPCDHGA9 expression was detected using quantitative real-time quantitative polymerase chain reaction (RT-qPCR) in 63 pairs of colorectal cancer tissues. Differential gene expression from high-throughput sequencing was analyzed using ingenuity pathway analysis (IPA) to explore the biological functions of PCDHGA9 and its potential regulated genes. Bioinformatics tools were employed to explore potential upstream regulatory microRNAs of PCDHGA9. Dual-luciferase assays were performed to demonstrate the regulation between PCDHGA9 and miR-1269a. Protein mass spectrometry suggested an interaction between PCDHGA9 and HOXA1. JASPAR predicted that HOXA1 may act as a transcription factor of CXCR4. Coimmunoprecipitation, dual-luciferase assays, and nuclear–cytoplasmic fractionation experiments confirmed the molecular mechanism involving PCDHGA9, CXCR4, HOXA1, and β-catenin. Transwell, wound healing, and western blot assays were conducted to confirm the impact of PCDHGA9, miR-1269a, and CXCR4 on the invasion, metastasis, and epithelial–mesenchymal transition (EMT) functions of CRC cells in in vitro experiments. A whole-body fluorescence imaging system was used to evaluate the combined impact of miR-1269a and PCDHGA9 on the invasion and metastasis of CRC in in vivo experiments.ResultsThe expression of PCDHGA9 was found to be lower in CRC tissues compared with their corresponding adjacent tissues. Low expression of PCDHGA9 potentially correlated with worse prognosis and increased chances of invasion and metastasis in CRC. miR-1269a was highly expressed in CRC tissues and acted as a negative regulator for PCDHGA9, promoting invasion, migration, and EMT of CRC cells. PCDHGA9’s interaction with HOXA1 downregulated CXCR4, a transcription factor, leading to accumulation of β-catenin and further promoting invasion, migration, and EMT of CRC cells.ConclusionsPCDHGA9, acting as a tumor suppressor, is downregulated by miR-1269a. The low level of PCDHGA9 activates the Wnt/β-catenin pathway by releasing its interaction with HOXA1, promoting the expression of CXCR4, and causing invasion, migration, and EMT in CRC.Graphical

BackgroundColorectal cancer (CRC), a complex and multifactorial disease, has been associated with elevated expression of microRNA miR-182-5p, although its precise regulatory role in CRC progression remains unclear. This study aims to identify potential therapeutic targets to improve clinical outcomes and to decipher the intricate role of miR-182-5p in the pathobiology of CRC.MethodsWe conducted comprehensive bioinformatics analyses using GEO databases to investigate differences in miRNA expression between CRC and normal tissues, with a particular focus on miR-182-5p. Its expression levels in CRC cells and tumor tissues were quantified by quantitative real-time PCR (qRT-PCR). The expression of neurocalcin delta (NCALD) and proteins related to Wnt/β-catenin signalling was evaluated by qRT-PCR and Western blotting. Pathological changes in tumor-bearing mice as well as the proliferation, invasion, and migration of CRC cells, were assessed. Tumor cell proliferation and apoptosis were examined using Ki-67 immunohistochemistry and TUNEL staining, respectively. A dual luciferase reporter assay explored the regulatory interaction between miR-182-5p and NCALD.ResultsOur findings reveal significantly elevated miR-182-5p levels in CRC tissues and cell lines, positively correlated with tumor invasion depth, differentiation degree, clinical stage, and lymph node metastasis. miR-182-5p appears to accelerate CRC progression in both cell lines and mouse models by downregulating NCALD, thereby enhancing Wnt/β-catenin signalling. This study identifies miR-182-5p as a pivotal enhancer of CRC progression, modulating Wnt/β-catenin signalling via NCALD regulation.ConclusionsThe findings position the miR-182-5p/NCALD axis as promising targets for CRC therapy, offering new avenues for treatment strategies.Trial registration: Retrospectively registered.

BackgroundColorectal cancer (CRC) represents a common malignancy in gastrointestinal tract. Iodine-125 (125I) seed implantation is an emerging treatment technology for unresectable tumors. This study investigated the mechanism of 125I seed in the function of CRC cells.MethodsThe CRC cells were irradiated with different doses of 125I seed (0.4, 0.6 and 0.8 mCi). miR-615 expression in CRC tissues and adjacent tissues was detected by RT-qPCR. miR-615 expression was intervened with miR-615 mimic or miR-615 inhibitor, and then the CRC cells were treated with 5-AZA (methylation inhibitor). The CRC cell growth, invasion and apoptosis were measured. The methylation level of miR-615 promoter region was detected. The xenograft tumor model irradiated by 125I seed was established in nude mice. The methylation of miR-615, Ki67 expression and CRC cell apoptosis were detected.Results125I seed irradiation repressed the growth and facilitated apoptosis of CRC cells in a dose-dependent manner. Compared with adjacent tissues, miR-615 expression in CRC tissues was downregulated and miR-615 was poorly expressed in CRC cells. Overexpression of miR-615 suppressed the growth of CRC cells. 125I seed-irradiated CRC cells showed increased miR-615 expression, reduced growth rate and enhanced apoptosis. The methylation level of miR-615 promoter region in CRC cells was decreased after 125I seed treatment. In vivo experiments confirmed that 125I seed-irradiated xenograft tumors showed reduced methylation of the miR-615 promoter and increased miR-615 expression, as well as decreased Ki67 expression and enhanced apoptosis. The target genes of miR-615 and its regulatory downstream pathway were further predicted by bioinformatics analysis.Conclusions125I seed repressed the growth and facilitated the apoptosis of CRC cells by suppressing the methylation of the miR-615 promoter and thus activating miR-615 expression. The possible mechanism was that miR-615-5p targeted MAPK13, thus affecting the MAPK pathway and the progression of CRC.

PurposeColorectal cancer (CRC) is the third most common cancer in the world. The purpose of this study was to investigate the role of TNNT2 in the proliferation, migration and invasion of CRC cells and its expression in CRC tissues to better understand the regulatory role of TNNT2 in CRC.MethodsWestern blotting (WB) and qPCR were used to detect the expression of TNNT2 in colorectal cancer tissues and paracancerous tissues. CCK-8, colony formation, Transwell and other experiments were used to clarify the role of TNNT2 in the proliferation, migration and invasion of colorectal cancer cells. Changes in TNNT2, EGFR and HER2 mRNA transcription levels were detected by SYBR Real-Time PCR assay, and the effects of TNNT2 overexpression or knockdown on the expression of EGFR, HER2 and EMT-related proteins in CRC cells were determined by WB. TNNT2 and EGFR intreaction was carried out in HCT116 cells by coimmunoprecipitation experiments.ResultsThe protein and mRNA expression level of TNNT2 in CRC tissues were higher than those in paracancerous tissues. The CCK-8 results suggested that overexpression of TNNT2 significantly promoted the proliferation of HCT116 and RKO cells, and TNNT2 konckdown gets the opposite result; and the colony formation results were the same as tthose of CCK-8 assay. Transwell invasion and migration experiments showed that overexpression of TNNT2 promoted the migration and invasion of HCT116 and PKO cells, and TNNT2 konckdown suppressed the migration and invasion of the these cells. The SYBR Green I real-time PCR method revealed that them RNA levels of TNNT2, EGFR and HER2 in the TNNT2 overexpression group were higher than those in RKO cells. WB showed that overexpressing TNNT2 increased the expression of EGFR and HER2 in HCT16 and RKO cells,decreased the expression of EMT marker E-cadherin, and increased the expression of Vimentin and N-cadherin. Konckdown of TNNT2 decreased the expression of EGFR and HER2, increased the expression of E-cadherin, and decreased the expression of Vimentin and N-cadherin in HCT16 and RKO cells. The immunocoprecipitation experiment showed that there was an interaction between EGFR and TNNT2.ConclusionTNNT2 can promote the proliferation, invasion and metastasis of colorectal cancer cells. There is an interaction between TNNT2 and EGFR protein. TNNT2 can upregulate EGFR and HER2-related proteins in colorectal cancer cells and promote the occurrence of EMT. Therefore, TNNT2 can promote the invasion and metastasis of CRC cells through the EGFR/HER2/EMT signal axis, suggesting that TNNT2 is a potential target of CRC treatment.

BackgroundColorectal cancer (CRC) is a tumor with high incidence and poor prognosis. An increasing number of studies have shown that intermediate filament proteins, such as nestin, participate in the regulation of tumor progression. However, the mechanism related to CRC is complex, and the role and underlying mechanism of nestin have not been elucidated in CRC.MethodsWe conducted quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blot analyses to examine the mRNA and protein levels in CRC and normal tissues. siRNAs targeting Nestin were transfected into CRC cells and then cell counting kit-8 (CCK-8), 5-ethynyl-2-deoxyuridine (EdU), sphere formation, and transwell analyses were used to assess the role of nestin in the proliferation, stem activity, migration, and invasive ability of CRC cells. Afterwards, nestin was overexpressed in CRC cells and P53 was overexpressed as a rescue group. CCK-8, EdU dyeing, sphere formation, and transwell assay was used to evaluated the role of Nestin/p53 axis in CRC cells.ResultsWe found high nestin expression and low p53 expression in CRC tissues and cells. Functionally, silencing of nestin suppressed the multiplication, stemness, and metastatic ability of Caco-2 and RKO cells. Encouragingly, rescue experiments suggested that overexpression of p53 partly restored the impacts of nestin overexpression on the viability, proliferation, and metastatic ability of CRC cells.ConclusionsWe confirmed that nestin and p53 play a functional role in the progression of CRC, and they may act as potential therapeutic targets for CRC treatment.

IntroductionColorectal cancer (CRC) remains one of the most lethal malignancies globally. Chemoresistance or reduced chemosensitivity is a key factor contributing to treatment failure in CRC, particularly in patients with advanced-stage disease. Iroquois homeobox protein 4 (IRX4), a transcription factor expressed in multiple tissues, has been demonstrated to be implicated in the progression of many cancers. Based on this background, IRX4 identified through Illumina Infinium 935K methylation chip analysis was chosen to explore its potential role in CRC.MethodsFollowing initial screening via the Illumina Infinium 935K methylation chip, we further confirmed the hypermethylation of IRX4 in CRC tissues using pyrosequencing technology. The expression levels of IRX4 were determined by immunofluorescence (IF) staining, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and western blot (WB) assays. In vitro cell experiments evaluated the effects of IRX4 overexpression on the proliferation, invasion, migration, and apoptosis of CRC cells, as well as its impact on the chemosensitivity to oxaliplatin (OXA). Mechanistically, after IRX4 overexpression, the phosphorylation level of epidermal growth factor receptor (EGFR) in CRC cells was detected. Luciferase reporter gene assays and immunofluorescence staining were used to investigate the effects of IRX4 overexpression on the activation of the nuclear factor-κB (NF-κB) pathway and the nuclear translocation of the p65 subunit.ResultsIRX4 was downregulated in CRC tissues and cell lines, and its expression was negatively correlated with tumor invasion depth and poor prognosis. In vitro experiments demonstrated that the overexpression of IRX4 inhibited CRC cell proliferation, migration, and invasion and promoted apoptosis. Moreover, IRX4 overexpression enhanced the chemosensitivity of CRC cells to OXA. Mechanistically, IRX4 overexpression significantly inhibited the TNF-α-induced NF-κB transcriptional activity and suppressed the nuclear translocation of NF-κB p65 in CRC cells. Furthermore, combined treatment with OXA and IRX4 overexpression significantly reduced the levels of EGFR and the phosphorylationlevels of EGFR downstream signaling molecules.ConclusionThese findings suggest that IRX4 may be a prognostic biomarker and therapeutic target in CRC. Moreover, IRX4 might regulate CRC progression and chemosensitivity by inhibiting the NF-κB /EGFR pathway, suggesting its potential as a therapeutic target to improve chemotherapeutic efficacy in CRC.

BACKGROUNDColorectal cancer (CRC) is one of the most common and fatal cancers worldwide. Synaptophysin-like 2 (SYPL2) is a neuroendocrine-related protein highly expressed in skeletal muscle and the tongue. The involvement of SYPL2 in CRC, including its level of expression and function, has not been evaluated.AIMTo evaluate the correlations of SYPL2 expression with lymph node metastasis (LNM) and prognosis in patients with CRC.METHODSThe levels of expression of SYPL2 in CRC and normal colorectal tissues were analyzed in multiple public and online databases. The associations between clinical variables and SYPL2 expression were evaluated statistically, and the associations between SYPL2 expression and prognosis in patients with CRC were analyzed using the Kaplan-Meier method and univariate/multivariate Cox regression analyses. SYPL2 expression was assessed in 20 paired CRC tissue and adjacent normal colorectal tissue samples obtained from Fuyang People’s Hospital, and the associations between SYPL2 expression and the clinical characteristics of these patients were investigated. Correlations between the levels of expression of SYPL2 and key targeted genes were determined by Pearson’s correlation analysis. The distribution of immune cells in these samples was calculated using the CIBERSORT algorithm. Gene set enrichment analysis (GSEA) was performed to evaluate the biofunction and pathways of SYPL2 in CRC.RESULTSSYPL2 expression was significantly lower in CRC tissue samples than in normal colorectal tissue samples (P < 0.05). High SYPL2 levels in CRC tissues correlated significantly with LNM (P < 0.05) and a poorer patient prognosis, including significantly shorter overall survival (OS) [hazard ratio (HR) = 1.9, P < 0.05] and disease-free survival (HR = 1.6, P < 0.05). High SYPL2 expression was an independent risk factor for OS in both univariate (HR = 2.078, P = 0.014) and multivariate (HR = 1.754, P = 0.018) Cox regression analyses. In addition, SYPL2 expression correlated significantly with the expression of KDR (P < 0.0001, r = 0.47) and the BRAFV600E mutation (P < 0.05). Higher SYPL2 expression was associated with the enrichment of CD8 T-cells and M0 macrophages in the tumor microenvironment. GSEA revealed that SYPL2 was associated with the regulation of epithelial cell migration, vasculature development, pathways in cancer, and several vital tumor-related pathways.CONCLUSIONSYPL2 expression was lower in CRC tissue than in normal colorectal tissue. Higher SYPL2 expression in CRC was significantly associated with LNM and poorer survival.

BackgroundColorectal cancer (CRC) is common and deadly, often leading to metastasis, challenging treatment, and poor outcomes. Understanding its molecular basis is crucial for developing effective therapies.AimsThis study aimed to investigate the role of Myosin Heavy Chain 11 (MYH11) in CRC progression, especially its effects on epithelial-mesenchymal transition (EMT) and cell behavior, and to explore its potential regulation by the EMT transcription factor zinc finger E-box binding homeobox 1 (ZEB1).MethodsDifferential expression analysis was performed in the GSE123390 and TCGA-READ datasets, and 317 intersection genes were identified. The hub gene MYH11 was identified based on Protein-protein interaction (PPI) analysis and expression validation. The effects of MYH11 and the EMT transcription factor (ZEB1) on the behavior of CRC cells were investigated in vitro.ResultsBioinformatics research revealed that MYH11 was considerably downregulated in CRC samples as compared to normal samples. Overexpression of MYH11 inhibited the proliferation, migration, and invasion of CRC cells. Western blotting (WB) testing showed that MYH11 overexpression inhibited EMT by elevating E-cadherin levels while suppressing ZEB1, vimentin, and N-cadherin expressions. By contrast, overexpression of ZEB1 promoted EMT and enhanced migration, invasion, and proliferation of CRC cells. The negative impacts of MYH11 affecting EMT markers and cell behaviors were partially mitigated by co-overexpression of MYH11 and ZEB1, indicating that MYH11 regulates EMT and CRC progression through ZEB1.ConclusionOur study shows MYH11 curbs CRC growth by blocking EMT and invasion, but ZEB1 overexpression reduces this effect. It uncovers key CRC pathways and suggests MYH11’s therapeutic potential.