Centromere Protein M Research Articles

TSC/mTORC1 mediates mTORC2/AKT1 signaling in c-MYC-induced murine hepatocarcinogenesis via centromere protein M.

Activated mTORC2/AKT signaling plays a role in hepatocellular carcinoma (HCC). Research has shown that TSC/mTORC1 and FOXO1 are distinct downstream effectors of AKT signaling in liver regeneration and metabolism. However, the mechanisms by which these pathways mediate mTORC2/AKT activation in HCC are not yet fully understood. Amplification and activation of c-MYC is a key molecular event in HCC. In this study, we explored the roles of TSC/mTORC1 and FOXO1 as downstream effectors of mTORC2/AKT1 in c-MYC-induced hepatocarcinogenesis. Using various genetic approaches in mice, we found that manipulating the FOXO pathway had minimal impact on c-MYC-induced HCC. In contrast, loss of mTORC2 inhibited c-MYC-induced HCC, an effect that was completely reversed by ablating TSC2, which activated mTORC1. Additionally, we discovered that p70/RPS6 and 4EBP1/eIF4E act downstream of mTORC1, regulating distinct molecular pathways. Notably, the 4EBP1/eIF4E cascade is crucial for cell proliferation and glycolysis in c-MYC-induced HCC. We also identified centromere protein M (CENPM) as a downstream target of the TSC2/mTORC1 pathway in c-MYC-driven hepatocarcinogenesis, and its ablation entirely inhibited c-MYC-dependent HCC formation. Our findings demonstrate that the TSC/mTORC1/CENPM pathway, rather than the FOXO cascade, is the primary signaling pathway regulating c-MYC-driven hepatocarcinogenesis. Targeting CENPM holds therapeutic potential for treating c-MYC-driven HCC.

Upregulation of CENPM promotes breast carcinogenesis by altering immune infiltration

BackgroundThe involvement of centromere protein M (CENPM) in various types of cancer has been established, however, its impact on breast cancer and immune infiltration remains unknown.MethodsWe examined the expression of CENPM in different cancer types by utilizing the Cancer Genome Atlas (TCGA) and Genotype Tissue Expression Pan-Cancer (GEO) databases. Using data from the TCGA, we examined the correlation between the expression of CENPM, the prognosis, and the clinicopathological features of individuals diagnosed with breast cancer. We conducted an enrichment analysis of CENPM using the clusterProfiler R software tool, utilizing data obtained from breast cancer patients and specimens at our institution. In addition to examining the correlation between CENPM expression and genes associated with immune checkpoints, the TIDE algorithm was employed to explore the potential of CENPM as a biomarker for immunotherapy in breast cancer. The impact of CENPM on the growth of breast cancer cells was evaluated through the utilization of the CCK8 test and the colony formation assay. The effect of CENPM on the migration of breast cancer cells was assessed using scratch and transwell assays.ResultsResearch findings indicate that elevated levels of CENPM are linked to patient outcomes in breast cancer and various clinicopathological features. Furthermore, elevated levels of CENPM expression correlated with decreased levels of CD8 + T cells and mast cells, increased levels of Tregs and Th2, and reduced levels of CD8 + T cells. Additionally, the coexpression of CENPM with the majority of genes related to immune checkpoints indicates its potential to forecast the effectiveness of treatment in breast cancer. Suppression of CENPM hampers the growth and movement of breast tumor cells.ConclusionsIn summary, our study findings indicate that CENPM may serve as a cancer-causing gene in breast cancer and also as a biomarker for predicting the efficacy of immunotherapy.The oncogene CENPM is associated with breast cancer and is involved in cell proliferation and immune infiltration.

The prognostic value and mechanisms of centromere protein M in patients with lung adenocarcinoma.

Centromere protein M (CENPM) has been reported to exert important roles in promoting tumor initiation and progression. However, the expression, effect, impact on prognosis and underlying mechanism of CENPM in lung adenocarcinoma (LUAD) remain unclear. Seventy-eight paired clinical samples of LUAD and corresponding adjacent non-tumor (ANT) tissues were obtained. The clinical pathological data and clinical outcome were tested, including univariate and multivariate Cox regression model. The relationship between CENPM expression and LUAD prognosis were identified according to the data obtained from the Cancer Genome Atlas (TCGA) database. Then, we explored the protein and mRNA levels of CENPM in LUAD and paired ANT tissues, and analyzed the correlation between CENPM and LUAD overall survival in our patients. In vitro studies, LUAD cell lines were treated with CENPM-short hairpin RNA (shRNA) (shCENPM), or transfected with CENPM overexpression plasmids with or without LY294002 (PI3K inhibitor) treatment. Cell proliferation ability was determined through cell counting kit-8 (CCK-8) assays. Cell cycle and apoptosis were detected by flow cytometer. The migration and invasion ability were assessed through Transwell assay. In vivo studies, the growth of xenografts in nude mice were evaluated after shCENPM stimulated cells injection, and the proliferation and apoptosis of xenografts were also analyzed. CENPM was significantly upregulated in LUAD patients compared with healthy controls, and CENPM upregulation was relevant to the higher pathological stages and poor survival rates in our LUAD patients. The bioinformatics analysis also revealed similar trends. CENPM could promote cell proliferation, cause alterations in cell cycle progression, enhance cell migration and invasion capacity, promote apoptosis in LUAD cell lines and promote the growth of xenografts in nude mice via regulation of AKT1/mTOR signaling pathway. CENPM was upregulated in LUAD patients, and it correlated with higher pathological stages and poor survival rates. CENPM could affect cell proliferation, cell cycle, cell migration and invasion capacity, and apoptosis in LUAD cell lines via regulation of AKT1/mTOR signaling pathway.

Upregulation of CENPM facilitates lung adenocarcinoma progression via PI3K/AKT/mTOR signaling pathway.

Centromere protein M (CENPM) is essential for chromosome separation during mitosis. However, its roles in lung adenocarcinoma (LUAD) progression and metastasis remain unknown. In this study, we aimed to explore the effects of CENPM on LUAD progression as well as the underlying mechanisms. We analyzed the expression of CENPM and its correlation with clinicopathological characteristics using GEO LUAD chip datasets and TCGA dataset. We further investigated the impact of CENPM on LUAD and . In silico analysis and qRT-PCR revealed that CENPM is upregulated in LUAD compared with that in normal lung tissues. Via gain/loss-of-function assays, we further found that CENPM promotes the LUAD cell cycle, cell proliferation, migration and invasion, and inhibits cell apoptosis. The study showed that loss of CENPM inhibits the growth of A549 xenografts. Furthermore, we found that CENPM can promote the phosphorylation of mTOR rather than directly affect the mTOR content. Inhibition of mTOR activity abrogates the promoting effects of CENPM on cell cycle progression, cell proliferation, migration and invasion. Taken together, these results show that CENPM plays an important role in the growth and metastasis of LUAD and may be a promising therapeutic target in LUAD.

Upregulation of centromere proteinM promotes tumorigenesis: A potential predictive target for cancer in humans.

Centromere protein M (CENPM), a protein required for chromosome separation, is involved in in mitosis. However, little has been reported about the roles of CENPM in various types of cancer. The present study identified that the mRNA expression levels of CENPM were significantly upregulated in 14 types of human cancer and identified a positive association between CENPM mRNA expression and patient mortality using the Oncomine, Gene Expression Profiling Interactive Analysis, Human Protein Atlas and Kaplan-Meier Plotter databases. A protein interaction network constructed with CENPM-interacting genes obtained from the cBioPortal demonstrated that nine genes participating in the cell cycle served key roles in the function of CENPM. Cell cycle analysis, reverse transcription-quantitative polymerase chain reaction, a Cell Counting Kit-8-based proliferation assay and a terminal deoxynucleotidyl transferase dUTP nick end labelling assay further revealed the tumorigenic and carcinogenic roles of CENPM in vitro. In addition, it was identified that the mRNA expression levels of five of the nine identified genes were significantly associated with CENPM in MCF7 cells and that CENPM was rarely mutated among various types of human cancer. In conclusion, the data from the present study revealed that CENPM exerted its pro-tumorigenic function by regulating cell cycle-associated protein expression and suggested that CENPM could be used as a prognostic marker for breast cancer.

LncRNA HCG18 contributes to the progression of hepatocellular carcinoma via miR-214-3p/CENPM axis.

Long non-coding RNA (lnc) HCG18 has been reported to contribute progression of a variety of tumours. However, its roles in hepatocellular carcinoma (HCC) remains unknown. In the current study, we intended to uncover the biological functions of HCG18 in HCC. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to detect the expression of HCG18, microRNA-214-3p (miR-214-3p) and centromere protein M (CENPM) messenger RNA (mRNA). The role of HCG18 in the growth and migration were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, wound healing assay and flow cytometry in vitro and animal experiments in vivo. The results showed that HCG18 was highly expressed in HCC tissues. HCG18 silencing inhibited the proliferation and migration while induced the apoptosis of HCC cells. Besides, miR-214-3p was down-regulated in HCC cells. Further experiments revealed that miR-214-3p could directly bind to HCG18 and exerted an anti-tumour role to counteracted siHCG18-1-mediated influence in HCC cells. Moreover, miR-214-3p could directly interact with CENPM mRNA and down-regulating the expression of CENPM. While HCG18 could up-regulate the expression of CENPM through acting as a sponge of miR-214-3p. Therefore, those results suggested HCG18 functioned as an oncogene to promote the proliferation and migration of HCC cells via miR-214-3p/CENPM axis.

Upregulation of CENPM facilitates tumor metastasis via the mTOR/p70S6K signaling pathway in pancreatic cancer.

Pancreatic cancer is a severe disease with high morbidity and mortality. However, the primary molecular mechanisms of pancreatic tumor formation and progression remain unclear. The present study using sequencing technology revealed that the centromere protein M (CENPM) gene was overexpressed in pancreatic cancer tissues. CENPM is one of the components of a complex that plays a central role in kinetochore protein assembly, mitotic progression and chromosome segregation. However, the biological function of CENPM in pancreatic cancer has yet to be determined. Hence, two effective siRNAs were designed to knock down CENPM. Notably, downregulation of CENPM inhibited pancreatic cancer cell proliferation, altered the cell cycle and limited pancreatic cancer cell migration and invasion via the mTOR/p70S6K signaling pathway. This research provides new evidence that CENPM overexpression plays a significant role in the progression of pancreatic cancer. Overall, the present findings indicated that CENPM may be a significant biomarker for predicting the development and progression of pancreatic malignancy.

Upregulation of CENPM promotes hepatocarcinogenesis through mutiple mechanisms

BackgroundHepatocellular carcinoma (HCC) still remains a dominating medical challenge in early diagnosis and clinical therapy. Centromere protein M (CENPM) has been proved to be over-expressed in HCC tissues, but carcinogenic mechanism of CENPM contributing to liver cancer is poorly understood.MethodsIn this study, we first explored mRNA and protein levels of CENPM in HCC samples, matching adjacent non-tumor tissues and six hepatoma cell lines by polymerase chain reaction (PCR), western blotting and immunohistochemistry (IHC). Clinical data of HCC patients downloaded from The Cancer Genome Atlas (TCGA) were also analyzed. The character of CENPM concerned with HCC progression through several functional experimentations in vitro and in vivo was researched. Bioinformatics was carried out to further discover biological functions of CENPM.ResultsCENPM was positively up-regulated in HCC and connected with a poor prognosis. Silencing CENPM repressed cell proliferation in vivo and in vitro, and knock-down CENPM inhibited cell migration and invasion. Additionally, depletion of CENPM can promote cell apoptosis and arrested cell cycle. Furthermore, single-gene gene set enrichment analysis (GSEA) analysis indicated that CENPM was linked to the P53 signaling pathway and cell cycle pathway, and our research supported this prediction. Finally, we also found that miR-1270 was a negative regulator and participated in post-transcriptional regulation of CENPM, and hepatitis B virus X protein (HBx) can promote hepatocellular carcinoma by suppressing miR1270.ConclusionCENPM was closely associated with HCC progression and it could be considered as a new possible biomarker along with a therapeutic target for HCC.

Identification of key candidate genes involved in melanoma metastasis

Metastasis is the most lethal stage of cancer progression. The present study aimed to investigate the underlying molecular mechanisms of melanoma metastasis using bioinformatics. Using the microarray dataset GSE8401 from the Gene Expression Omnibus database, which included 52 biopsy specimens from patients with melanoma metastasis and 31 biopsy specimens from patients with primary melanoma, differentially expressed genes (DEGs) were identified, subsequent to data preprocessing with the affy package, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. A protein-protein interaction (PPI) network was constructed. Mutated genes were analyzed with 80 mutated cases with melanoma from The Cancer Genome Atlas. The overall survival of key candidate DEGs, which were within a filtering of degree >30 criteria in the PPI network and involved three or more KEGG signaling pathways, and genes with a high mutation frequency were delineated. The expression analysis of key candidate DEGs, mutant genes and their associated genes were performed on UALCAN. Of the 1,187 DEGs obtained, 505 were upregulated and 682 were downregulated. ‘Extracellular exosome’ processes, the ‘amoebiasis’ pathway, the ‘ECM-receptor interaction’ pathway and the ‘focal adhesion’ signaling pathway were significantly enriched and identified as important processes or signaling pathways. The overall survival analysis of phosphoinositide-3-kinase regulator subunit 3 (PIK3R3), centromere protein M (CENPM), aurora kinase A (AURKA), laminin subunit α 1 (LAMA1), proliferating cell nuclear antigen (PCNA), adenylate cyclase 1 (ADCY1), BUB1 mitotic checkpoint serine/threonine kinase (BUB1), NDC80 kinetochore complex component (NDC80) and protein kinase C α (PRKCA) in DEGs was statistically significant. Mutation gene analysis identified that BRCA1-associated protein 1 (BAP1) had a higher mutation frequency and survival analysis, and its associated genes in the BAP1-associated PPI network, including ASXL transcriptional regulator 1 (ASXL1), proteasome 26S subunit, non-ATPase 3 (PSMD3), proteasome 26S subunit, non ATPase 11 (PSMD11) and ubiquitin C (UBC), were statistically significantly associated with the overall survival of patients with melanoma. The expression levels of PRKCA, BUB1, BAP1 and ASXL1 were significantly different between primary melanoma and metastatic melanoma. Based on the present study, ‘extracellular exosome’ processes, ‘amoebiasis’ pathways, ‘ECM-receptor interaction’ pathways and ‘focal adhesion’ signaling pathways may be important in the formation of metastases from melanoma. The involved genes, including PIK3R3, CENPM, AURKA, LAMA1, PCNA, ADCY1, BUB1, NDC80 and PRKCA, and mutation associated genes, including BAP1, ASXL1, PSMD3, PSMD11 and UBC, may serve important roles in metastases of melanoma.

SU110 - ROLE OF 108 SCHIZOPHRENIA-ASSOCIATED LOCI IN MODULATING PSYCHOPATOLOGICAL DIMENSIONS IN SCHIZOPHRENIA AND BIPOLAR DISORDER

Background Schizophrenia is a highly heritable disorder and the Schizophrenia Working Group of the Psychiatric Genomics Consortium (PGC) identified 108 loci with genome-wide association with broad schizophrenia diagnosis. Schizophrenia is a clinically complex entity and the role of these 108 loci in modulating specific clinical features of the disease is still unknown. This study investigated which symptom dimensions may be affected by these loci in schizophrenia and bipolar disorder. Methods Positive, negative and depressive symptoms, suicidal ideation, cognition, violent behaviors, quality of life and early onset were investigated in the Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) and Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD) samples. Only white subjects were included to avoid possible confounding from ancestry stratification. Imputation of genotypes was carried out after standard quality control using the Haplotype Reference Consortium panel as reference. Individual loci were investigated using linear or logistic regression, then genes within 50 Kbp from polymorphisms with p Results 89 polymorphisms among the 108 of interest were available in both samples, 479 and 810 white subjects were included from CATIE and STEP-BD, respectively. rs75059851 (IGSF9B gene) was associated with negative symptoms in CATIE (p=0.00048); no other variants survived after multiple-testing correction. rs1023500 (CENPM gene) showed a trend of association with early onset with consistent direction in both samples (p=0.033 in STEP-BD and p=0.073 in CATIE). GeneMania analysis showed that negative symptoms and suicidal ideation had a relevant enrichment of Gene Ontology (GO) terms related to acetylcholine neurotransmission, monoamines and ion channel activity while pyridoxal phosphate binding and fucose metabolism showed enrichment for cognition. GO terms referred to processes involved in extracellular matrix structure were found enriched for early onset. PRS showed nominal associations with violent behaviors (p=0.034, Nagelkerke's R2=0.014) and depressive symptoms (p=0.04, R2=0.009). Discussion This study provided preliminary evidence that a schizophrenia-associated variant in the IGSF9B (immunoglobulin superfamily member 9B) gene may modulate negative symptoms. IGSF9B is involved in GABAergic signaling and it may mediate negative symptoms by affecting depression and cognition. A polymorphism in the CENPM (centromere protein M) gene showed an interesting trend of association with early onset. CENPM encodes for an inner protein of the kinetochore, the multi-protein complex that binds spindle microtubules to regulate chromosome segregation during cell division. Multi-locus models may provide interesting insights about the biological mechanisms that medicate specific symptom domains and the genetic overlap between schizophrenia and bipolar disorder.

Characterization of CENPM Isoform Expression, Intracellular Localization, and Potential Functional Interactions In Mature B-Cell Malignancies

AbstractAbstract 2496The lymphoid-specific PANE1 human minor histocompatibility antigen (mHAg) is overexpressed in primary chronic lymphocytic leukemia (CLL) cells. The PANE1 mHAg peptide is encoded by an alternative transcript of the centromere protein M (CENPM) gene, and may be a desirable target for immunotherapy due to its lymphoid specificity, and hence its potential to instigate a graft-versus-leukemia effect with absent or mitigated graft-versus-host disease. While its immunotherapeutic potential is compelling, little is presently known about the expression, cellular localization, and functional role of CENPM isoforms. To gain further insight into CENPM isoform expression and function in B-lymphoid and other malignancies, we assessed these characteristics of the CENPM transcript encoding a 58 amino acid isoform encompassing the mHAg (referred to as CENPM-S), and the longer canonical 180 amino acid CENPM isoform (referred to as CENPM-L). These two isoforms share a common 46 amino acid C-terminus, and differ in that the N-terminal 134 amino acids of CENPM-L are substituted in CENPM-S by a 12 amino acid sequence containing the mHAg peptide, which is derived from a novel exonization event within the fifth intron of the canonical genomic sequence. The CENPM-L isoform has been shown to be a critical component of the centromeric protein complex on which the transient assembly of the kinetochore occurs during mitosis. We determined that the novel CENPM-S mHAg-encoding exon and its adjacent predicted alternative promoter (distinct from that of CENPM-L) is derived from a LINE2 non-LTR retrotransposon insertion. To better assess the types of lymphoid malignancies that could potentially serve as targets of a CENPM-S-specific immune response, we examined a diverse panel of primary CLL cells, lymphoid malignancies and lymphoma cell lines for the expression of CENPM-S and CENPM-L via quantitative real-time PCR (qRT-PCR). In accord with our previous results, highest levels of CENPM-S were observed in primary CLL cells. Within lymphoma cell lines, moderate levels of CENPM-S were seen in mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL) cells, and expression was detectable in only one out of four multiple myeloma cell lines surveyed. Within a panel of human lymphoma tumor samples, highest levels of CENPM-S were observed in marginal zone lymphoma, DLBCL, MCL, follicular lymphoma, and one peripheral T-cell lymphoma sample. Strikingly, the highest levels of CENPM-S and the lowest levels of CENPM-L were always observed specifically in primary CLL cells, and such high ratios of CENPM-S to CENPM-L expression were not observed in any other B-cell malignancy surveyed. These results suggest a profound transcriptional switch from the upstream canonical promoter to the putative LINE2 element-proximal CENPM-S alternative promoter, which occurs exclusively in CLL cells. In order to further elucidate the cellular localization and potential function of the CENPM-S and CENPM-L isoforms, we transiently transfected vectors encoding FLAG-tagged CENPM isoforms in HeLa cells. Transfected cells were subjected to subcellular fractionation followed by Western blot with anti-FLAG antibodies, which indicated CENPM-L localization to cytoplasmic, membrane, soluble nuclear, and cytoskeletal fractions, in accord with our observations of endogenous CENPM-L in Granta-519 MCL cells using CENPM-L-specific monoclonal antibody. In contrast, CENPM-S exhibited localization in cytoplasm and membrane and was present only weakly in the soluble nuclear fraction. Preliminary immunofluorescence microscopy data are in agreement with these patterns of intracellular localization. We are presently assessing CENPM isoforms for impact on cell cycle, proliferation, and apoptosis, in order to better understand their potential involvement in biological processes underlying CLL and other non-Hodgkins lymphomas. To further elucidate the potential functional role of CENPM-S in mature B-lymphocytes and mature B-lymphoid malignancies, we performed yeast two-hybrid library screening with a human bone marrow cDNA library to reveal putative interacting proteins. The non-muscle myosin protein, myosin IIA, was revealed as a strong candidate CENPM-S interacting protein. We have recently validated via sandwich ELISA an interaction between endogenous myosin IIA (within Granta-519 MCL cell extract) and recombinant CENPM-S. Disclosures:No relevant conflicts of interest to declare.