Novel Biomarker For Colorectal Cancer Research Articles

Investigating the Effects of AL049796.1 Silencing in Inhibiting High Glucose-Induced Colorectal Cancer Progression.

Patients with colorectal cancer (CRC) and diabetes share many risk factors. Despite a strong association between diabetes and CRC being widely studied and confirmed, further genetic research is needed. This study found higher AL049796.1 and TEA domain transcription factor 1 (TEAD1) levels (both mRNA and protein) in CRC tissues of diabetic patients compared with nondiabetics, but no significant difference in miR-200b-3p levels. A positive correlation between AL049796.1 and TEAD1 protein existed regardless of diabetes status, whereas miR-200b-3p was only negatively correlated with TEAD1 protein in nondiabetic CRC tissues. In vitro experiments have shown that high glucose (HG) treatment increased AL049796.1 in CRC cells, and AL049796.1 silencing reduced HG-induced proliferation, migration and invasion, as well as connective tissue growth factor, cysteine-rich angiogenic inducer 61, and epidermal growth factor receptor protein expression. Mechanistic investigations indicated that AL049796.1 could mitigate suppression of miR-200b-3p on TEAD1 posttranscriptionally by acting as a competitive binder. In vivo, subcutaneous CRC tumors in streptozotocin (STZ)-induced mice grew significantly faster; AL049796.1 silencing did not affect the growth of subcutaneous CRC tumors but significantly reduced that of STZ-induced mice. Our study suggests that AL049796.1 independently contributes to the risk of CRC in diabetic patients, highlighting its potential as both a therapeutic target and a novel biomarker for CRC among individuals with diabetes.

Collagen synthase P4HA3 as a novel biomarker for colorectal cancer correlates with prognosis and immune infiltration

ObjectiveIn this study, we aimed to determine whether proly4-hydroxylase-III (P4HA3) could be used as a biomarker for the diagnosis of colorectal cancer (CRC) as well as for determining prognosis. MethodsWe used The Cancer Genome Atlas (TCGA) database to analyze P4HA3 expression in CRC and further investigated the association between P4HA3 and clinicopathological parameters, immune infiltration, and prognosis of patients with CRC. Enrichment analysis was conducted to investigate the potential biological role of P4HA3 in CRC. To verify the results of TCGA analysis, we performed immunohistochemical staining of 180 clinical CRC tissue samples to probe into the relationship of P4HA3 expression with lymphocyte infiltration and immune checkpoints expression. ResultsThe expression of P4HA3 was significantly higher in CRC tissues and associated with a higher degree of malignancy and poorer prognosis in CRC. The results of enrichment analysis indicated that P4HA3 may be associated with the epithelial-mesenchymal transition process and the immune response. Immunohistochemical staining results showed that high P4HA3 expression was associated with high infiltration levels of CD8+ and Foxp3+ TILs and high PD-1/PD- L1 expression. Lastly, patients with CRC co-expressing P4HA3 and PD-1 had a significantly worse prognosis. ConclusionHigh expression of P4HA3 is associated with adverse clinical features and immune cell infiltration in CRC, and has the potential to serve as a biomarker for predicting CRC prognosis.

Identification of EGR4 as a prospective target for inhibiting tumor cell proliferation and a novel biomarker in colorectal cancer.

EGR4 (Early Growth Response 4) is a member of the EGR family, involving in tumorigenesis. However, the function and action mechanism of EGR4 in the pathogenesis of colorectal cancer (CRC) remain unclear. To address this, we assessed the prognosis of CRC based on EGR4 using the Kaplan-Meier plotter tool and tissue microarray. The abundance of immunoinfiltration was evaluated through ssGSEA, TISIDB, and TIMER. In vitro experiments involving knockdown or overexpression of EGR4 were performed, and RNA-sequencing was conducted to explore potential mechanisms. Furthermore, we used oxaliplatin and 5-fluorouracil to validate the impact of EGR4 on chemo-resistance. Pan-cancer analysis and tissue microarray showed that EGR4 was highly expressed in CRC and significantly correlated with an unfavorable prognosis. Moreover, EGR4 expression was associated with immunoinfiltration and cancer-associated fibroblasts in the CRC microenvironment. Functional enrichment demonstrated that high-expressional EGR4 were involved in chromatin and nucleosome assembly. Additionally, EGR4 promoted the proliferation of CRC cells. Mechanistically, EGR4 upregulated TNFα to activate the NF-κB signaling pathway, and its knockdown reduced p65 nuclear translocation. Importantly, combining shEGR4 with oxaliplatin and 5-fluorouracil significantly inhibited CRC proliferation. Taken together, these findings provide new insights into the potential prognosis and therapeutic targets of EGR4 in CRC.

Prediction and validation of circulating G-quadruplexes as a novel biomarker in colorectal cancer.

There are some problems in the clinical diagnosis of colorectal cancer (CRC), such as the difficulty in saving samples, so it is the most popular research work to develop a diagnostic index and method that is easy to obtain, convenient to save and stable. G-quadruplex (G4) is a unique structure found in DNA and it plays a crucial biological role in tumor formation. G4 is derived from DNA with good stability, and the DNA of serum samples is easy to obtain. Therefore, G4 has the potential as an ideal marker for CRC diagnosis. However, it has not received more attention. Through bioinformatics-based G4 mutation prediction in the genome, we discovered that the G4 quantity in SW480 cells was lower than that of the reference gene. However, it was unclear how the G4 quantity changed in the actual samples. We detected the G4 content by fluorescence in cells and human serum samples. G4 content was significantly higher than that in NCM480 (P<0.001). To further explore the relationship between tumorigenesis and G4, we knocked out the TP53 gene in SW480 cells and found that the G4 content decreased significantly (64%) (P<0.001). The difference in G4 content is a key factor in distinguishing between normal and tumor cells. Furthermore, we detected G4 in serum samples from 27 healthy individuals and 27 patients with CRC and found that G4 was significantly increased in those with CRC (P<0.001) by 1.94 fold. We also evaluated the G4 model using receiver operating characteristic (ROC), with an area under the curve of 0.91, and found it to have excellent specificity and sensitivity. The increased G4 is an important characteristic in patients with CRC and has clinical application value as a novel biomarker. The relationship between G4 and TP53 regulation may be a potential target for future cancer studies, and as attention to this area of research increases, the underlying mechanisms may be better understood, potentially benefiting clinical cancer treatment.

NDUFA4L2 is a novel biomarker for colorectal cancer through bioinformatics analysis.

Colorectal cancer (CRC) is a major cause of cancer-related deaths worldwide. NDUFAL42 is an important mitochondrial respiratory chain subunit that plays a critical role in cellular energy metabolism. However, the role of NDUFA4L2 in CRC remains unclear. Therefore, we used the data obtained from The Cancer Genome Atlas (TCGA) database to prove the relationship between NDUFA4L2 and CRC. The expression levels of NDUFA4L2 in CRC tissues were analyzed by immunohistochemical staining of NDUFA4L2 from the HPA database. Wilcoxon rank sum test, Chi-square test, Fisher exact test and logistic regression were used to evaluate relationships between clinical-pathologic features and NDUFA4L2 expression. Receiver operating characteristic (ROC) curves were used to describe binary classifier value of NDUFA4L2 using area under curve (AUC) score. Kaplan-Meier method and Cox regression analysis were used to evaluate factors contributing to prognosis. Gene oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were used to predict the function of differentially expressed genes associated with NDUFA4L2. Gene set enrichment analysis (GSEA) was used to predict canonical pathways associated with NDUFA4L2.Immune infiltration analysis was performed to identify the significantly involved functions of NDUFA4L2. Protein-protein interaction (PPI) networks were established and 20 hub genes identified with Cytoscape software. Increased NDUFA4L2 expression in CRC was associated with T stage (P = .019), N stage (P < .001), Pathologic stage (P = .020), Residual tumor (P = .023), Perineural invasion (P = .039), Lymphatic invasion (P = .007), Histological type(P < .001), PFI event (P = .007) and DSS event (P = .004).ROC curve suggested the significant diagnostic and prognostic ability of NDUFA4L2 (AUC = 0.878). High NDUFA4L2 expression predicted a poorer Overall-survival (P = .021), poorer progression-free interval (P = .001), and poorer Disease Specific Survival (P = .002). GO, KEGG, GSEA and immune infiltration analysis showed that NDUFA4L2 expression was correlated with regulating the function of DNA and some types of immune infiltrating cells. NDUFA4L2 expression was significantly correlated with poor survival and immune infiltrations in CRC, and it may be a promising prognostic biomarker in CRC.

F-Box and Leucine-Rich Repeat Protein 7 Is a Prognostic Biomarker and Is Correlated with the Immunosuppressive Microenvironment in Colorectal Cancer.

Background: Colorectal cancer (CRC) is a common malignancy of the digestive system, but its specific mechanisms of occurrence and development remain incompletely understood. F-Box and leucine-rich repeat protein 7 (FBXL7) is a subunit of the Skp-cullin-F-box ubiquitin ligase, involved in cell cycle regulation, endothelial cell damage, and inflammatory immunological responses. However, the role of FBXL7 in CRC remains unknown. In this study, we investigated the clinical significance and potential mechanism of FBXL7 expression in CRC progression. Methods: We utilized data from The Cancer Genome Atlas (TCGA) and the University of California Santa Cruz Xena (UCSC Xena) database for bioinformatic analyses. Clinical CRC samples were used to confirm FBXL7 expression. Gene set enrichment analysis (GSEA) and various databases, such as TCGA, UCSC Xena, cBioPortal, University of ALabama at Birmingham CANcer data analysis portal, MethSurv, Tumor Immune Estimation Resource (TIMER), TIMER2.0, Tumor-Immune System Interaction Database, and Tumor Immune Dysfunction and Exclusion Database (TIDB), were used to investigate the role of FBXL7 in CRC. Statistical analysis was performed using R (v.3.6.3) or GraphPad Prism 8.0. Results: Our findings revealed the predictive significance of FBXL7 in CRC patients. FBXL7 expression was associated with tumor stage, lymph node stage, pathological stage, perineural invasion, and lymphatic invasion. GSEA analysis identified associations between FBXL7 and extracellular matrix organization, as well as immune-related pathways. Immunological analysis revealed a correlation between high FBXL7 expression and the development of an immunosuppressive microenvironment. Conclusion: Identifying FBXL7 as a novel biomarker for CRC could shed light on the promotion of CRC development by the immune environment.

Identification of lncRNA PCAT19 as potential novel biomarker for colorectal cancer

Long non-coding RNAs have been implicated in biological processes, and are dysregulated in types of cancer. Studies have shown that PCAT19 and CKMT2-AS1 lncRNAs promote tumor growth, invasion, and metastasis by regulating signaling pathways and modulating the gene expression. This study investigated the expression levels of lncRNAs PCAT19 and CKMT2-AS1 in colorectal tumors and normal tissues. First, Using GEPIA2 database, we compared the expression level of target lncRNAs between primary colon adenocarcinoma tumor and normal tissues. Then, the expression levels of lncRNAs PCAT19 and CKMT2-AS1 were detected in 35 colorectal tumors and paired adjacent tissues using qRT-PCR. A receiver operating characteristic (ROC) curve was used to evaluate the value of these lncRNAs as biomarkers. Statistical analysis based on GEPIA2 showed that both lncRNAs PCAT19 and CKMT2-AS1 were significantly decreased in colon adenocarcinoma compared to the normal group (P < 0.001). Experimental analysis showed that the expression level of lncRNA PCAT19 was decreased in colorectal tumors (p < 0.0001) compared to normal tissues. While the expression level of lncRNA CKMT2-AS1 did not change in tumor tissues, it decreased in non-metastatic tumors compared to normal tissues (p = 0.04). The significantly downregulation of lncRNA PCAT19 expression in both metastatic and non-metastatic colorectal tumors compared to normal tissue suggests that PCAT19 may play a role in the carcinogenesis and progression of colorectal cancer and may provide potential therapeutic targets for colorectal cancer. Based on the results of ROC curve analysis, lncRNA PCAT19 may also serves as a novel potential good biomarker in diagnosis colorectal cancer (AUC = 0.94, p < 0.0001) but no significant was found for lncRNA CKMT2-AS1 (p > 0.05).

Identification of ZMYND19 as a novel biomarker of colorectal cancer: RNA-sequencing and machine learning analysis.

Colorectal cancer (CRC) is the third most common cause of cancer-related deaths. The five-year relative survival rate for CRC is estimated to be approximately 90% for patients diagnosed with early stages and 14% for those diagnosed at an advanced stages of disease, respectively. Hence, the development of accurate prognostic markers is required. Bioinformatics enables the identification of dysregulated pathways and novel biomarkers. RNA expression profiling was performed in CRC patients from the TCGA database using a Machine Learning approach to identify differential expression genes (DEGs). Survival curves were assessed using Kaplan-Meier analysis to identify prognostic biomarkers. Furthermore, the molecular pathways, protein-protein interaction, the co-expression of DEGs, and the correlation between DEGs and clinical data have been evaluated. The diagnostic markers were then determined based on machine learning analysis. The results indicated that key upregulated genes are associated with the RNA processing and heterocycle metabolic process, including C10orf2, NOP2, DKC1, BYSL, RRP12, PUS7, MTHFD1L, and PPAT. Furthermore, the survival analysis identified NOP58, OSBPL3, DNAJC2, and ZMYND19 as prognostic markers. The combineROC curve analysis indicated that the combination of C10orf2-PPAT- ZMYND19 can be considered as diagnostic markers with sensitivity, specificity, and AUC values of 0.98, 1.00, and 0.99, respectively. Eventually, ZMYND19 gene was validated in CRC patients. In conclusion, novel biomarkers of CRC have been identified that may be a promising strategy for early diagnosis, potential treatment, and better prognosis.

PLA2G12A as a Novel Biomarker for Colorectal Cancer with Prognostic Relevance

Metastasis is the leading cause of colorectal cancer (CRC)-related deaths. Therefore, the identification of accurate biomarkers predictive of metastasis is needed to better stratify high-risk patients to provide preferred management and reduce mortality. In this study, we identified 13 new genes that modified circulating tumor cell numbers using a genome-wide genetic screen in a whole animal CRC model. Candidate genes were subsequently evaluated at the gene expression level in both an internal human CRC cohort of 153 patients and an independent cohort from the TCGA including 592 patients. Interestingly, the expression of one candidate, PLA2G12A, significantly correlated with both the time to recurrence and overall survival in our CRC cohort, with its low expression being an indicator of a poor clinical outcome. By examining the TCGA cohort, we also found that low expression of PLA2G12A was significantly enriched in epithelial–mesenchymal transition signatures. Finally, the candidate functionality was validated in vitro using three different colon cancer cell lines, revealing that PLA2G12A deficiency increases cell proliferation, migration, and invasion. Overall, our study identifies PLA2G12A as a prognostic biomarker of early-stage CRC, providing evidence that its deficiency promotes tumor growth and dissemination.

Plasma cfDNA role in detecting cardiovascular, metabolic and other diseases in a FIT+ population enrolled for a colorectal cancer screening program.

e15541 * Francesco Pepe made an equal contribution to the study Background: Cell-free DNA (cfDNA) has long been established as a useful tool in a variety of clinical settings, including infectious, auto-immune, cardiovascular and neoplastic diseases. In a study that is conducted at ASL NA 3 SUD (Naples, Italy) with the University of Naples Federico II, cfDNA was used to conduct mutational analysis using a novel biomarker for colorectal cancer. In this study, the observation of cfDNA value has suggested its potential use as a disease detector. This abstract reports findings of a secondary analysis in the study. Methods: 667 FIT+ subjects, aged 50-74, participating in a colorectal cancer screening program enrolled in a clinical trial focusing on the role of a novel biomarker in colorectal cancer diagnosis. The subjects' medical history included cardiovascular diseases (CVD) and diabetes primarily, collectively named as Concomitant Diseases (CD). 398 patients had at least one CD, while 269 had none. A whole blood sample was obtained from each study subjects and total plasma cfDNA was measured using QuantiDNA™ test, manufactured by DiaCarta Inc. ROC curve was run to assess diagnostic efficacy and determine cutoff points of cfDNA for detecting the CD and specifically cardiovascular ones. Logistic regression was used to determine the probability of disease in subjects with an elevated cfDNA concentration. Adjusted estimates were obtained to assess the potential confounding effect of CD vs colorectal neoplasia (CN). Results: The area under the curve (AUC) for cfDNA as a predictor of CD was 0.562 (95% C.I. 0.518, 0.607); the cutoff point was 13.44 ng/ml, with a sensitivity of 50% (95% C.I 45%, 55%) and a specificity of 63% (95% C.I. 57%, 69%). The logistic regression to assess the probability of CD showed an odds ratio (OR) for cfDNA of 1.6279 (95% C.I. 1.19, 2.234; p = 0.0023) that is, one unit increase in cfDNA will increase the odds of disease by ~63%. A specific assessment was made for cfDNA as a detector for CVD. The AUC was 0.567 (95% C.I. 0.523, 0.611), the cutoff point was 12.8 ng/ml, with a sensitivity of 55% (95% C.I. 50%, 60%) and a specificity of 57% (95% C.I. 51%, 63%). The logistic regression indicated an OR of 1.681 (95% C.I. 1.2378, 2.2876; p &lt; 0.001) meaning that a one unit increase in cfDNA will increase the odds of disease by ~68.1%. Lastly, the logistic regression of cfDNA as a predictor of CN adjusted for CD showed a change in slope &gt; 10% from the unadjusted form, suggesting a potential confounding effect. Conclusions: Although the study was not designed to assess the validity of cfDNA as a detector of diseases, these secondary findings suggest that cfDNA can be used as a disease detector for CD including CVD, which encourages further studies. Attention should be directed towards potential confounding effects, such as from CD or CN since cfDNA can be elevated by several different conditions.

Prediction and Validation of Circulating G-quadruplexes as a Novel Biomarker in Colorectal Cancer

Prediction and Validation of Circulating G-quadruplexes as a Novel Biomarker in Colorectal Cancer

A Positive Feedback Loop of E2F4-Mediated Activation of MNX1 Regulates Tumour Progression in Colorectal Cancer.

Purpose: Colorectal cancer (CRC) is the 3rd most prevalent malignant tumour globally. Although significant strides have been made in diagnosis and treatment, its prognosis at the moment remains unpromising. Therefore, there is an urgent and desperate need to identify novel biomarkers of CRC and evaluate its mechanism of tumourigenesis and development. Methods: JASPAR and RNAinter databases are used to analyze target genes associated with colorectal cancer. Western blotting, q-PCR and immunohistochemistry et, al. were used to detect the level of MNX1 in patients with colorectal cancer, and Chip-PCR was used to detect the targeted binding ability of E2F4 and MNX1. The cells and animal models overexpressed MNX1 and E2F4 were constructed by shRNA transfection. Results: Herein, MNX1 was highly expressed and linked to favourable overall survival curves in colorectal cancer. The functional assay revealed that MNX1 overexpression could promote proliferation, migration, and invasion of CRC cells. Based on the prediction of the JASPAR and RNAinter databases, the transcription factor, E2F4, was bound to the MNX1 promoter region. The Chromatin Immunoprecipitation (ChIP) assay verified the interactions between MNX1 and E2F4 in CRC. Additionally, we found that sh-E2F4 markedly downregulated the MNX1 levels and reduced CRC progression in vivo and in vitro, which reversed MNX1 overexpression. Conclusion: Therefore, our research discovered that E2F4-mediated abnormal MNX1 expression promotes CRC progression and could become a novel diagnostic or therapeutic target of CRC.

VASN promotes colorectal cancer progression by activating the YAP/TAZ and AKT signaling pathways via YAP.

Colorectal cancer (CRC) is one of the most common gastrointestinal malignancies. Vasorin (VASN) has been reported to be critical in tumor development and angiogenesis. However, VASN has not been reported in CRC, and its role is unclear. In this study, VASN expression is upregulated in CRC compared with the normal tissues, and VASN expression positively correlates with N stage and poor overall survival by analysis of different datasets and 32 CRC clinicopathologic samples. Overexpression of VASN significantly promotes CRC cell progression, including proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while knockdown of VASN inhibits CRC progression. We found that VASN was associated with the YAP/TAZ and PI3K/AKT pathways by gene set enrichment analysis (GSEA) and gene ontology (GO) analysis. Notably, western blotting, immunofluorescence staining and co-immunofluorescence (co-IP) confirmed that VASN could interact with YAP and activate the YAP/TAZ and PTEN/PI3K/AKT pathways, and knockdown of YAP reversed this effect. Importantly, our findings indicate that VASN interacts with YAP to inhibit YAP phosphorylation and stimulates CRC proliferation, migration, and invasion through activation of the YAP/TAZ-TEAD target gene CTGF and PTEN/PI3K/AKT pathways. Our results also show that knockdown of YAP reverses the cellular phenotype induced by increased VASN. In conclusion, our study reveals that VASN acts as an oncogene to stimulate tumor progression in CRC, providing new insights into the molecular mechanisms of CRC development and representing a possible novel biomarker for CRC.

N6-methyladenosine-modified circular RNA QSOX1 promotes colorectal cancer resistance to anti-CTLA-4 therapy through induction of intratumoral regulatory T cells.

Colorectal cancer (CRC) is the 3rd most common cancer worldwide. CircRNAs are promising novel biomarkers for CRC. T regulatory (Treg) cells express the immune checkpoint receptor of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) and promote tumor immunological tolerance. We therefore investigate the biological functions and mechanisms of circQSOX1 in CRC tumorigenesis; involvement of circQSOX1 in promoting Treg cell-mediated CRC immune escape in anti-CTLA-4 therapy. Bioinformatics analyses were performed for circQSOX1expressions, specific binding sites, and N6-methyladenosine (m6A) motifs of circQSOX1, thatwere further validated with a series of experiments. Functions of circQSOX1 in promoting CRC development, Treg cells-based immune escape, and anti-CTLA-4 therapy response were investigated both in vitro and in vivo. High circQSOX1 expression was associated with carcinogenesis and poor clinical outcome of CRC patients. METTL3-mediated RNA m6A modification on circQSOX1 could be read by IGF2BP2 in CRC cells. CircQSOX1 promoted CRC development by regulating miR-326/miR-330-5p/PGAM1 axis. CircQSOX1 regulated glycolysis and promoted immune escape of CRC cells, and inhibits anti-CTLA-4 therapy response in CRC patients. m6A-modified circQSOX1 facilitated CRC tumorigenesis by sponging miR-326 and miR-330-5p to promotes PGAM1 expression, which further promoted CRC immune escape by activating glycolysis and inactivating the anti-CTLA-4 therapy response of CRC. Combined treatment with sh-circQSOX1 and anti-CTLA-4 could be a strategy to overcome Treg cell-mediated CRC immune therapy resistance.

An integrated lab-on-a-chip platform for pre-concentration and detection of colorectal cancer exosomes using anti-CD63 aptamer as a recognition element

Colorectal cancer (CC) is one of the common causes of cancer-related deaths around the globe. Identification of a novel biomarker for CC is of paramount importance for early diagnostics and reducing its mortality. Among the most promising biomarker candidates, exosomes hold great potential for cancer diagnosis, management, and treatment. Exosomes are extracellular vesicles secreted from cells and they contribute to the intercellular communication, immune response and the pathogenesis of many diseases including cardiovascular diseases, neurodegenerative diseases, and cancer. Several methods have been developed/utilized for exosome isolation and purification. However, these methods are time-consuming and have low purification efficiency. In this work, we developed the “apta-magnetic biosensor” platform for isolation, purification and detection of exosomes from cell culture. Anti-CD63 aptamer, which is conjugated to the surface of magnetic nanobeads, was used as a recognition element. A dynamic separation system was used which employs a transverse magnetic field along a microfluidic channel. The channel was exposed to an alternate magnetic field which imposes alternate magnetic force onto the magnetic bead-exosome complex. The combination of the fluid flow and magnetic force generates several “alternate trapping and releasing” events under continuous-flow conditions with each event representing a washing cycle. The graphene coated onto the surface of the magnetic nanobeads was used as a quencher for the fluorescently labeled aptamer (OFF state) in the absence of the target. Upon the addition of CD63 target protein, the aptamer dissociates from the graphene and binds to the target hence increasing the fluorescence intensity (ON state). A calibration plot of variable concentrations of exosomes vs fluorescence intensity was obtained and the detection limit was calculated as 1457 particles/mL. The specificity of the sensor was tested using closely associated proteins. The results showed that the aptamagnetic isolation, pre-concentration of exosomes and quantification demonstrate great potential for various clinical applications.

Long non-coding RNA AC087388.1 as a novel biomarker in colorectal cancer

BackgroundSeveral investigations have reported diverse roles of long non-coding RNA (lncRNA) in biological processes, tumor development, and progression of colorectal cancer (CRC). In this study, we investigated the lncRNA AC087388.1 tumorigenic role in CRC cells.MethodsThe CRC tissues were collected at the Reza Radiotherapy and Oncology Center, Mashhad, Iran. The human SW-48 and HT-29 CRC cell lines were obtained from the national cell bank of Iran. The cells were cultured according to ATCC (the American Type Culture Collection) recommendations. Quantitative real-time PCR was applied to assess the RNA expression. ShRNA transfection was done to downregulate the target gene. MTT and apoptosis assays were conducted to evaluate cell proliferation and viability, respectively. Colony formation assay, wound healing assay, and invasion assay were applied to determine growth, motility, and invasion of the cells, respectively. ENCORI online tool was used as downstream enrichment analysis.ResultsForty CRC patients were encompassed in this study. The results demonstrated that the lncRNA SLC16A1-AS1, AC087388.1, and ELFN1-AS1 were significantly overexpressed in the CRC tissues in comparison to their normal counterpart margins. All the lncRNAs have shown significant Area Under Curve (AUC) values in the patients. Downregulation of lncRNA AC087388.1 remarkably decreased the cell proliferation and viability of the CRC cells. In addition, the data demonstrated that the downregulation of lncRNA AC087388.1 significantly suppressed cell growth and colony formation capability in the cells. Also, downregulation of lncRNA AC087388.1 attenuated motility and invasion of CRC cells, and significantly decreased the expression of invasion genes. In-silico functional enrichment analysis indicated that the lncRNA AC087388.1 has contributed to crucial signaling pathways in tumorigenesis such as the p53 and Wnt signaling pathways, apoptosis, and cell cycle.ConclusionsAltogether, we showed that lncRNA AC087388.1 has an oncogenic role in tumorigenesis of CRC, and it can be considered as a novel diagnostic and prognostic biomarker in CRC.

MiR-3614-5p Is a Potential Novel Biomarker for Colorectal Cancer.

MiR-3614-5p has been found in a variety of cancers including colorectal cancer. However, the association of miR-3614-5p with colorectal cancer is still unclear. Based on the Cancer Genome Atlas (TCGA) database, the relationship between miR-3614-5p and colorectal cancer can be proved. Wilcoxon rank-sum test was used to compare the miR-3614-5p expression in colorectal cancer tissues and under normal conditions, respectively. The logistic regression method was further employed to analyze the relationship between miR-3614-5p and clinicopathological characteristics. Also, the correlation between miR-3614-5p and survival rate was evaluated by Kaplan-Meier and Cox regression analysis. Besides, gene set enrichment analysis (GSEA) was used to investigate the biological functions of miR-3614-5p. The decrease of miR-3614-5p expression of colorectal cancer was significantly correlated with N stage (OR) = 0.7 for N1&N2 vs. N0), M stage (OR = 0.5 for M1 vs. M0), pathologic stage (OR = 0.7 for Stage III & Stage IV vs. Stage I & Stage II), neoplasm type (OR = 0.5 for rectum adenocarcinoma vs. colon adenocarcinoma), and lymphatic invasion (OR = 0.6 for YES vs. NO) (all p-values < 0.05). Kaplan-Meier survival analysis showed that colorectal cancer with low miR-3614-5p has a poorer prognosis than that of high miR-3614-5p (p = 0.005). According to univariate analysis, low miR-3614-5p was associated with poor overall survival (OS) [hazard ratio (HR) = 0.599; 95% confidence interval (CI): 0.418-0.857; p = 0.005]. In multivariate analysis, miR-3614-5p was closely related to OS (HR = 0.630; 95% CI: 0.405-0.978, p = 0.021). GSEA showed that the high expression phenotype of miR-3614-5p differentially enriches the P53 pathway. Meanwhile, the high expression phenotype of miR-3614-5p enhanced NK T cell activation, negative T cell selection, response to interleukin 2, and response to tumor cells. MiR-3614-5p is a possible prognostic marker of low survival rate for patients with colorectal cancer. Moreover, the P53 pathway and P38MAPK pathway may be the key pathways regulated by miR-3614-5p in colorectal cancer.

ASPM is a Novel Candidate Gene Associated with Colorectal Cancer Cell Growth.

Colorectal cancer (CRC) is one of the most prevalent diseases worldwide; however, the molecular mechanisms involved in CRC remain unclear. Thus, we aimed to explore a novel biomarker for CRC. In this study, we screened 361 differentially expressed genes; 152 downregulated genes; and 209 upregulated genes) through analysis of the GSE44861, GSE110223, GSE110224, and GSE113513 CRC datasets. Next, ASPM, CCNA2, CCNB1, CEP55, KIF20A, MAD2L1, MELK, RRM2, TOP2A, TPX2, TRIP13, and TTK were identified as hub genes associated with the cell cycle in CRC through comprehensive bioinformatics analysis using the Cytoscape and Metascape software, the Database for Annotation, Visualization, and Integrated Discovery (DAVID), and the Oncomine and Gene Expression Profiling Interactive Analysis 2 (GEPIA2) databases. Furthermore, ASPM mRNA expression in CRC tissues was verified in Oncomine, The Cancer Genome Atlas and our data, and ASPM was found to be significantly upregulated in CRC tissues compared with that in the noncancer colon tissues. Functionally, we showed that overexpression of ASPM significantly promoted the proliferation and inhibited apoptosis; silencing of ASPM suppressed the proliferation of CRC cells by affecting the cell cycle G1/S transition by reducing cyclin E1 expression, and inducing apoptosis. Overall, our findings indicated that ASPM plays a crucial role in the regulation of CRC cell proliferation, and ASPM is a potential candidate diagnostic tool and therapeutic target for CRC.

MiR-1539 and Its Potential Role as a Novel Biomarker for Colorectal Cancer.

PurposeWe investigated microRNA (miR) 1539 as a potential biomarker for predicting the risk and pathobiological behavior of colorectal cancer (CRC).MethodsOur strategy consisted of analyzing 100 serum samples from 51 CRC patients, 49 healthy controls (HCs), and another 56 CRC tissue and matched normal adjacent to tumor (NAT) samples. The relative expression levels of miR-1539 in exosomes, serum and tissues were detected and compared in the different groups, using reverse transcription-polymerase chain reaction (RT-qPCR). The diagnostic value and potential function of miR-1539 were investigated using clinicopathological data combined with bioinformatics analysis.ResultsMiR-1539 expression was significantly up-regulated in exosomes (p = 0.003) and cancer tissue (p < 0.001) from CRC patients. MiR-1539 expression levels in serum varied according to different tumor sites (right-sided vs. left-sided, p = 0.047; left-side CRC vs. HCs, p = 0.031). In terms of diagnostic efficacy, miR-1539 expression in exosomes may help distinguish CRC cases from HCs with a sensitivity of 92.2%, and miR-1539 expression in serum may improve the specificity to 96.6% for left-sided CRC diagnosis. When combined with clinicopathological data, serum miR-1539 levels were positively associated with vascular endothelial growth factor (VEGF) expression (p = 0.028), whilst levels in CRC tissue were positively associated with increased Ki-67 levels (p = 0.035). Poorer pathologic differentiation was potentially related to an increased tendency of miR-1539 expression in CRC tissue (p = 0.071). Based on our bioinformatics analysis, miR-1539 may have a significant mechanistic influence on CRC genesis and progression.ConclusionsCirculating or tissue based miR-1539 may be used as a novel potential biomarker for CRC screening, and a predictor of poor clinicopathological behavior in tumors.

Volatile organic compounds analysis as a potential novel screening tool for colorectal cancer: A systematic review and meta-analysis.

The purpose of this meta-analysis was to assess the usefulness of volatile organic compounds (VOC) as a potential novel biomarker for colorectal cancer (CRC).We systematically searched PubMed, Embase, Web of Science, and Cochrane Library databases for observational studies (published before November 25th, 2019; no language restrictions) comparing the VOC analysis between patients with CRC and healthy controls. We evaluated the pooled sensitivity, specificity, diagnostic odds ratio, positive and negative likelihood ratio, as well as summary receiver operating characteristic curve and area under the curve.We identified a total of 10 observational studies that included 381 patients with CRC and 436 healthy controls. Bivariate analysis yielded a pooled sensitivity of 0.82 (95% confidence interval [CI] = 0.77-0.86), specificity of 0.79 (95% CI = 0.71-0.85), positive likelihood ratio of 3.8 (95% CI = 2.8-5.3), and negative likelihood ratio of 0.23 (95% CI = 0.17-0.30). The area under the curve was 0.87 (95% CI = 0.84-0.90). The pooled diagnostic odds ratio was 17 (95% CI = 10-28). Sensitivity analysis indicated that the pooled results were stabilized. The Deeks' funnel plot asymmetry test (P = .41) suggested no potential publication bias.Our pooled data confirmed the associations between VOC analysis and CRC, highlighting the usefulness of VOC analysis as a potential novel screening tool for CRC. However, standardization of VOC collection and analysis methods for CRC screening is required in future research.