Fig 1: The effect of ATF2/LNC000882/miR-214-3p/CENPM pathway on the binding of Tubulin to Centromere related DNA sequence. (A–E) The HepG2 cells were transfected with vectors and harvested for the ChIP assays. The complex of Tubulin with Centromere related DNA sequence was separated by the antibodies of Tubulins. (F) The expression level of ATF2, (G) LNC000882, (H) miR-214-3p or (I) CENPM in HepG2 cells were examined by the qPCR. The results were shown as the schematic-diagram images of the Centromere related DNA sequence (A), the images or mean ± SD from qPCR of the ChIP results (B–E) or the mean ± SD from the qPCR. *p < 0.05.
Fig 2: Knockdown of miR-214-3p effectively reversed LINC00882 knockdown-induced inhibition on HCC progression in vitro. (A) RT-PCR and Western blot examined the expression of CENPM in Huh7 and HepG2 cells transfected with sh-NC, sh-LINC00882-1, sh-LINC00882-1+ NC inhibitor or sh-LINC00882-1+ miR-214-3p inhibitor. CCK-8 assays (B), colony formation assays (C) and Transwell assays (D) of HepG2 and Huh7 cells after transfection. **p < 0.01.
Fig 3: CENPM was identified as a direct target of miR-214-3p in HCC cells. (A) Bioinformatics tools reveal the complementary binding sites within miR-214-3p and CENPM. (B) Pan-cancer analysis of CENPM using TCGA datasets. (C, D) The expression pattern of CENPM in HCC specimens with different stages. (E) Survival assays of HCC patients according to the expression of CENPM by analyzing TCGA datasets. (F) Luciferase reporter assay validated the molecular binding. (G) RT-PCR determined the expression of CENPM in Huh7 and HepG2 cells transfected with NC mimics, miR-214-3p mimics, NC inhibitors or miR-214-3p inhibitors. *p < 0.05, **p < 0.01, ***p < 0.001.
Fig 4: CENPM is upregulated in lung cancer and associated with the prognosis of LUAD patients via bioinformatics analysis. (A) CENPM gene expression across tumor types and normal tissues, including LUAD and LUSC from online TIMER database. (B) CENPM, CENPA, CENPF, CENPU, CENPO and CENPH gene expression upregulated in LUAD tissues compared with normal tissues. (C) CENPM gene expression upregulated in LUAD (TCGA, n=515) compared with normal tissues (GTEx, n=347). (D) CENPM expression in paired samples (n=57) in TCGA database. (E) ROC curve of CENPM expression for LUAD with AUC of 0.847. (F) Kaplan-Meier survival curves for OS and DSS of LUAD patients from TCGA database. (G) Kaplan-Meier survival curves for OS and FP of LUAD patients from Kaplan-Meier Plotter database. (H) The higher CENPM mRNA levels, the higher LUAD pathological stages from TCGA database. (I) GO functional analysis of CENPM in BP, MF and CC. KEGG pathway enrichment analysis of CENPM. *, P<0.05; **, P<0.01; ***, P<0.001. ACC, adrenocortical carcinoma; AUC, area under the curve; BLCA, bladder urothelial carcinoma; BP, biological process; BRCA, breast invasive carcinoma; CC, cellular component; CENP, centromere protein; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; CI, confidence interval; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B cell lymphoma; DSS, disease specific survival; ESCA, esophageal carcinoma; FPR, false positive rate; FP, first progression; GBM, glioblastoma multiforme; GO, Gene Ontology; GTEx, genotype-tissue expression; HNSC, head and neck carcinoma; HPV, human papillomavirus; HR, hazard ratio; KEGG, Kyoto Encyclopedia of Genes and Genomes; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LAML, acute myeloid leukemia; LGG, brain lower grade glioma; LIHC, liver hepatocellular carcinoma; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; MF, molecular function; OS, overall survival; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; ROC, receiver operative characteristic; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TCGA, The Cancer Genome Atlas; TGCT, testicular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; TPM, transcript per million; TPR, true positive rate; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma.
Fig 5: Upregulation of CENPM in LUAD patients correlates with higher pathological stages and poor patient survival. (A) CENPM protein level was increased in LUAD tissues compared with paired ANT, as measured by WB. Relative protein levels were determined after normalization to a-tubulin. (B) CENPM mRNA level was increased in LUAD tissues compared with paired ANT, as measured by RT-qPCR. The mRNA level was normalized to the mRNA level of GAPDH as an endogenous control. (C) Representative images of immunohistochemical normal lung tissues, low expression and high expression of CENPM expression in LUAD. (D) CENPM mRNA levels increased with clinical pathological stages. (E) Kaplan-Meier survival curves for OS in LUAD patients. Bars indicate the means ± SD. *, P<0.05; **, P<0.01; ****, P<0.0001. CENPM, centromere protein M; ANT, adjacent non-tumor tissues; LUAD, lung adenocarcinoma; T, tumor; HR, hazard ratio; WB, Western blot; RT-qPCR, real-time quantitative polymerase chain reaction; OS, overall survival; SD, standard deviation.
Supplier Page from Abcam for Anti-CENPM antibody