ABSTRACT
INTRODUCTION: The aim of the study was to investigate the effect of CNRIP1 promoter methylation on the proliferative, invasive and migration potential of colorectal cancer cells, including its potential use for the early detection and prognostic assessment of colorectal cancer. MATERIAL AND METHODS: Quantitative methylation-specific PCR (qMSP) was used to detect the methylation status of the CNRIP1 promoter region in peripheral blood samples drawn from patients with colorectal adenocarcinoma, benign colorectal adenoma, and matched healthy controls. Putative CpG methylation sites were then pyrosequenced. We subsequently suppressed CNRIP1 methylation within colon cancer cells via treatment with 5-azacytidine and overexpressed colon cancer cells by transfection with a CNRIP1-overexpression pcDNA3.0 plasmid. Thereafter, the CNRIP1 methylation status and mRNA and protein expressions levels were determined. Finally, the proliferative, invasive and migration abilities of cell lines were determined with the CCK-8 and Transwell cell assays. RESULTS: There were differences in the methylation status at loci 2216, 2226, 2231, 2245, and 2254 within the promoter region of CNRIP1 between patients with colorectal adenocarcinoma, colorectal adenoma, and healthy volunteers. The methylation status of CpG sequence 2245 significantly correlated with tumor diameter, invasion depth, TNM stage, grade, and lymph node metastasis (p < 0.05). The proliferative, invasive and migration abilities of colon cancer cells treated with 5-azaC or transfected with a CNRIP1-overexpression plasmid were significantly impaired relative to negative controls (p < 0.05). CONCLUSIONS: The methylation status at locus 2245 within the CNRIP1 promoter region has potential value for the early detection and prognostic evaluation of colorectal cancers. Demethylation of the CNRIP1 promoter or overexpression of CNRIP1 can reduce the proliferative and migration abilities of colon cancer cells.
ABSTRACT
BACKGROUND: Protein arginine methyltransferases 1 (PRMT1) is over-expressed in a variety of cancers, including lung cancer, and is correlated with a poor prognosis of tumor development. This study aimed to investigate the role of PRMT1 in nonsmall cell lung cancer (NSCLC) migration in vitro. METHODS: In this study, PRMT1 expression in the NSCLC cell line A549 was silenced using lentiviral vector-mediated short hairpin RNAs. Cell migration was measured using both scratch wound healing and transwell cell migration assays. The mRNA expression levels of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 1, 2 (TIMP1, 2) were measured using quantitative real-time reverse transcription-polymerase chain reaction. The expression levels of protein markers for epithelial-mesenchymal transition (EMT) (E-cadherin, N-cadherin), focal adhesion kinase (FAK), Src, AKT, and their corresponding phosphorylated states were detected by Western blot. RESULTS: Cell migration was significantly inhibited in the PRMT1 silenced group compared to the control group. The mRNA expression of MMP-2 decreased while TIMP1 and TIMP2 increased significantly. E-cadherin mRNA expression also increased while N-cadherin decreased. Only phosphorylated Src levels decreased in the silenced group while FAK or AKT remained unchanged. CONCLUSIONS: PRMT1-small hairpin RNA inhibits the migration abilities of NSCLC A549 cells by inhibiting EMT, extracellular matrix degradation, and Src phosphorylation in vitro.
Subject(s)
Carcinoma, Non-Small-Cell Lung/enzymology , Cell Movement/physiology , Epithelial-Mesenchymal Transition/physiology , Protein-Arginine N-Methyltransferases/genetics , Protein-Arginine N-Methyltransferases/metabolism , Blotting, Western , Carcinoma, Non-Small-Cell Lung/genetics , Cell Line , Cell Movement/genetics , Epithelial-Mesenchymal Transition/genetics , Extracellular Matrix Proteins/metabolism , Humans , RNA, Small Interfering/genetics , RNA, Small Interfering/physiologySubject(s)
Dexamethasone/therapeutic use , Drugs, Chinese Herbal/therapeutic use , Proctitis/drug therapy , Radiation Injuries/drug therapy , Rectum/drug effects , Administration, Rectal , Adult , Aged , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/therapeutic use , Dexamethasone/administration & dosage , Drug Therapy, Combination , Female , Humans , Middle Aged , Phytotherapy , Powders , Proctitis/etiology , Radiation Injuries/complications , Rectum/pathology , Rectum/radiation effects , Uterine Cervical Neoplasms/radiotherapyABSTRACT
OBJECTIVE: To investigate serum levels of endogenous oxidation agents, anti-oxidation agents and clinical significance in the patients with cerebral vascular disease (CVD). METHODS: Using biochemical methods, the levels of serum nitric oxide (NO), nitric oxide synthase (NOS), malondialdehyde (MDA) and anti-oxidants vitamin E (VitE), vitamin C (VitC), superoxide dismutase (SOD) in 49 patients with cerebral hemorrhage (CH), 65 patients with cerebral infarction(CI) and 35 patients with other nervous system diseases and 34 healthy controls were determined. RESULTS: In CH and CI groups, the levels of serum NO and MDA and the activity of serum NOS were significantly higher than that of the two other groups (P<0.05 or P<0.01). On the other hand, the patients with CH and CI had lower VitE, VitC levels and SOD activity than that of the two control groups (P<0.05 or P<0.01). CONCLUSION: These findings suggest NO and NOS plays an important role in pathogenesis of cerebral damage after CH and CI. Determination of the concentrations of NO, VitE, VitC, MDA level, and NOS and SOD activity in serum can also help judge the seriousness and the course of the disease.
