ART1 knockdown decreases the IL-6-induced proliferation of colorectal cancer cells.

Colorectal cancer (CRC) is a worldwide health concern. Chronic inflammation is a risk factor for CRC, and interleukin-6 (IL-6) plays a pivotal role in this process. Arginine-specific mono-ADP-ribosyltransferase-1 (ART1) positively regulates inflammatory cytokines. ART1 knockdown reduces the level of glycoprotein 130 (gp130), a key transducer in the IL-6 signalling pathway. However, the relationship between ART1 and IL-6 and the resulting effects on IL-6-induced proliferation in CRC cells remain unclear. The aims of this study were to investigate the effects of ART1 knockdown on IL-6-induced cell proliferation in vitro and use an in vivo murine model to observe the growth of transplanted tumours. The results showed that compared with the control, ART1-sh cancer cells induced by IL-6 exhibited reduced viability, a lower rate of colony formation, less DNA synthesis, decreased protein levels of gp130, c-Myc, cyclin D1, Bcl-xL, and a reduced p-STAT3/STAT3 ratio (P < 0.05). Moreover, mice transplanted with ART1-sh CT26 cells that had high levels of IL-6 displayed tumours with smaller volumes (P < 0.05). ART1 and gp130 were colocalized in CT26, LoVo and HCT116 cells, and their expression was positively correlated in human CRC tissues. Overall, ART1 may serve as a promising regulatory factor for IL-6 signalling and a potential therapeutic target for human CRC.

The prognostic value of RHBDF2 in Pan-Cancer, and its correlation with cell Adhesion of Hepatocellular Carcinoma.

The impact of RHBDF2 on the expression and potential function in many cancers is still unknown. Therefore, the expression and methylation modification of RHBDF2 were evaluated across TCGA cancers in this study. Moreover, two methods, COX regression and Kaplan-Meier, were utilized for analyses of the prognoses of RHBDF2 in patients. Besides, the association between RHBDF2 and immune microenvironment, mutation, tumor mutation burden and microsatellite instability was analyzed with Pearson correlation. We verified RHBDF2 expression in hepatocellular carcinoma (HCC) compared with normal cell and tissue samples, detected the effects of RHBDF2 knockdown on biological functions in HCC cells, and detected CD4, CD8 and CD68 expression in hepatocellular carcinoma tissues and paired normal tissues. Given these results, the significant mRNA overexpression and promoter hypomethylation of RHBDF2 in various tumor types was showed, and a clear relationship between RHBDF2 overexpression and unfavourable overall survival and progression-free survival was observed, including liver hepatocellular carcinoma (LIHC), glioma (GBMLGG) and pancreatic adenocarcinoma (PAAD). Additionally, hypomethylation of RHBDF2 can affect the overall survival in some tumors. Furthermore, a clear correlation between RHBDF2 and infiltration of immune cells, immune-related molecules, TMB and MSI was observed. Besides, RHBDF2 expression is upregulated in HCC cells and tissues, and RHBDF2 knockdown could decrease the cell adhesion ability of HCC cells. More importantly, the expression of CD4, CD8 and CD68 was higher in HCC tissues. Altogether, the research denoted that RHBDF2 can be a prognostic biomarker for cancers according to these results and participate in cell adhesion of HCC cells.

RHBDF2 is correlated with immune infiltrates in hepatocellular carcinoma and may have potential as a biomarker.

Immune checkpoint inhibitors have been approved for the treatment of advanced hepatocellular carcinoma (HCC). However, immunotherapy requires the identification of suitable biomarkers to guide treatment. The gene for rhomboid 5 homolog 2 (IRHom2), which encodes the rhombus protease iRhom2, activates the MAP3K7-dependent pathway and promotes hepatic steatosis. Thus, we hypothesized an involvement of this gene in HCC. We report that RHBDF2 expression is dramatically upregulated in HCC. RHBDF2 upregulation is associated with tumor stage, lymph node metastasis, tumor protein P53 mutation, and worse prognoses in HCC patients. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes and gene set enrichment analysis enrichment analysis indicated that RHBDF2 was involved in immune signal pathways. Moreover, RHBDF2 expression was positively related not only to immune infiltration, but also to immune cell markers. Examination of several types of HCC infiltrated by immune cells revealed that the group with high expression of RHBDF2 showed the worst outcomes. Therefore, RHBDF2 may have potential as a novel biomarker for predicting prognosis and is related to immune infiltrates in HCC.

An integrated analysis of Dynamin 1 Like: A new potential prognostic indicator in hepatocellular carcinoma.

Dynamin 1 Like (DNM1L), a member of dynamin superfamily capable of mediating mitochondrial outer membrane division, plays a key role in the progression of different types of tumors. However, the prognostic value, clinical significance of DNM1L and its specific mechanism involved in tumorigenesis of hepatocellular carcinoma (HCC) have not been investigated clearly. In this study, we found that the expression of DNM1L were significantly higher in HCC tissues than adjacent/normal liver tissues based on multiple data sets obtained from TCGA, GEO and ONCOMINE database, also its protein expression form Drp1 is significantly higher in HCC tissues than adjacent tissues, and is related to the degree of differentiation. Kaplan-Meier curves suggested that high DNM1L expression prominently correlated with poorer overall survival, progression-free survival, relapse-free survival and disease-specific survival. Multivariate analysis showed that higher DNM1L expression was independent prognostic factors of shorter overall survival and disease-free survival. Kyoto Encyclopedia of Genes and Genomes and Gene set enrichment analysis analysis combined with validation experiments revealed the regulatory role of DNM1L on key molecules in the metabolism of xenobiotics by cytochrome p450 pathway, and DNM1L may also affects invasion and metastasis capability of HCC by mediating extracellular matrix -receptor interaction pathway. Moreover, analysis showed that higher DNM1L, CYP2C9, CYP3A4, CYP1A2 expression were associated with the resistance to sorafenib therapy. TIMER and CIBERSORT analysis indicated that the increase of DNM1L expression may affect the infiltration of immune cells in the tumor microenvironment. Taken together, the above results indicated that DNM1L could be able to serve as a promising independent predictor and therapeutic target for HCC patients.

Identification of the role of mono-ADP-ribosylation in colorectal cancer by integrated transcriptome analysis.

Our previous study clarified the carcinogenic properties of arginine-specific mono-ADP-ribosyltransferase 1 (ART1), which results in a critical post-translational modification that changes the structure and function of proteins and is widely involved in important processes. This study provides, for the first time, a comprehensive transcriptomic analysis of colorectal cancer cells with ART1 silencing by Illumina RNA-Seq and related verification experiments. Lentiviral infection was used to construct a CT-26 cell line with stable knockdown of the ART1 gene, and a whole transcriptome sequencing technique was performed to identify differentially expressed genes (DEGs). GO and KEGG classification/enrichment analyses and verification experiments were performed to determine the role of ART1 in the progression of colorectal cancer. A total of 5552 DEGs, GO functions and KEGG pathways with the highest enrichment, various SNPs, and diverse splicing patterns were identified. Importantly, knockdown of ART1 affected the splicing of certain key genes related to tumor cell growth and downregulated the expression of the key gene PTBP1 for alternative splicing. The overall attenuation of the endoplasmic reticulum unfolded protein response (UPR) signaling pathway caused by the inhibition of mono-ADP-ribosylation of GRP78 could disrupt UPR signaling, leading to the occurrence of apoptosis to impede tumorigenesis. ART1, which is clustered in organelles, may promote the development of colorectal cancer by participating in a variety of new mechanisms, including endoplasmic reticulum stress regulation and alternative splicing, and may be a good clinical drug target for targeted therapy of CRC.

Preliminary Study on the Identification of BRAFV600E Mutation in Colorectal Cancer by Near-Infrared Spectroscopy.

INTRODUCTION: In metastatic colorectal cancer (mCRC), the B-type Raf kinase (BRAF)V600E mutation is a molecular biomarker of poor prognosis and is of great importance to drug target. Currently, the commonly used methods for detecting BRAFV600E mutation include immunohistochemistry (IHC) and gene sequencing, but both present certain limitations. Near-infrared (NIR) spectroscopy is a spectroscopy technology that takes advantage of the electromagnetic wavelength between visible light and mid-infrared light. METHODS: IHC was used to detect the expression of BRAFV600E protein with the BRAFV600E (VE1) antibody in 42 cases of paraffin-embedded (FFPE) mCRC tissue sections. The NIR-discriminant analysis model (NIRS-DA) was established using 6 cases of wild-type and 6 cases of mutant-type BRAF specimens. RESULTS: IHC detection results revealed 13 cases of weakly positive (+), 1 case of moderately positive (++), and 28 cases of negative (-) CRC. Compared with the next-generation sequencing (NGS) results, the positive rate was 66.7%. The classification accuracy of calibration (CAC) was 100% compared with the results of NGS, demonstrating that the BRAFV600E mutant NIRS-DA model, verified by 2 cases of wild-type and 2 cases of mutant-type CRC samples was established. The NIRS-DA model was used to predict gene mutation in the CRC samples, 7 cases were positive (+), and 35 cases were negative (-), and the classification accuracy of prediction (CAP) was 83.3% (35/42). DISCUSSION: The NIRS-DA model-predicted results were in high agreement with the detection results of NGS, and the difference in IHC is not statistically significant (P>0.05). However, this study is a preliminary discussion on a methodology due to its small sample size.