scholarly journals Research of the Methylation Status of miR-124a Gene Promoter among Rheumatoid Arthritis Patients

2013 ◽  
Vol 2013 ◽  
pp. 1-4 ◽  
Author(s):  
Qiao Zhou ◽  
Li Long ◽  
Guixiu Shi ◽  
Jing Zhang ◽  
Tong Wu ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Promoter Methylation ◽  
Methylation Status ◽  
Gene Promoter ◽  
Healthy Controls ◽  
Bisulfite Conversion ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Synovial Tissues

Objective. To analyze the methylation status of miR-124a loci in synovial tissues of rheumatoid arthritis (RA) patients using methylation-specific polymerase chain reaction (MSP).Materials and Methods. DNA obtained from the frozen tissue of 7 RA samples, 6 osteoarthritis (OA) samples, and 3 healthy controls were undergoing bisulfite conversion and then analyzed for miR-124a promoter methylation using MSP assay.Results. miR-124-a1 and miR-124-a2 promoter methylation were both seen in 71.4% of RA samples compared to 16.7% of OA samples. miR-124-a3 promoter methylation was seen in 57.1% of RA samples and 0% of OA samples. All the three loci were unmethylated in 3 healthy controls.Conclusion. The methylation status of miR-124a seen in this study concurs with that reported in tumor cells, indicating epigenetic dysregulation constituents, a mechanism in the development of rheumatoid arthritis.

scholarly journals Detection of SNCA and FBN1 Methylation in the Stool as a Biomarker for Colorectal Cancer

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Wen-han Li ◽  
Hao Zhang ◽  
Qi Guo ◽  
Xuan-di Wu ◽  
Zi-sen Xu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Methylation Status ◽  
Good Alternative ◽  
Noninvasive Detection ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Specific Polymerase Chain Reaction ◽  
Stool Samples ◽  
Polymerase Chain

Aim.We examined the methylation status of SNCA and FBN1 genes in patients’ paired tissue and stool samples for detection of colorectal cancer (CRC).Patients and Methods. 89 DNA tissue samples (normal/cancer) and corresponding stool samples were analyzed in our study. In addition, 30 stool samples were collected as healthy controls.Results. The methylation level of those samples was measured by methylation-specific polymerase chain reaction (MSP). The result shows that compared with the paired controls, both SNCA and FBN1 were significantly hypermethylated in CRC patients in tissue samples (P<0.001). In the stool samples, hypermethylated SNCA and FBN1 were detected to be significantly higher than that in normal stool samples (P<0.001). The combined sensitivity of at least one positive among the two markers in stool samples was 84.3%, with a specificity of 93.3%. In addition, our experiment suggested that the positive rates of SNCA and FBN1 in Dukes A stage were significantly higher than that of FOBT (P=0.039;P=0.006, resp.).Conclusion. We concluded that methylation testing of SNCA and FBN1 genes in stool sample may offer a good alternative in a simple, promising, and noninvasive detection of colorectal cancer.

scholarly journals MiR-498 suppresses proliferation and inflammation in fibroblast-like synoviocytes in rheumatoid arthritis via targeting JAK1

2021 ◽  
Vol 18 (10) ◽  
pp. 2011-2017
Author(s):  
Lan Chai ◽  
Xian Zhen Zhang ◽  
Hai fang Ma ◽  
Fang Yuan
Keyword(s):  
Rheumatoid Arthritis ◽  
Cell Cycle ◽  
Polymerase Chain Reaction ◽  
Therapeutic Target ◽  
Cell Apoptosis ◽  
Inflammatory Responses ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Synovial Tissues ◽  
Fibroblast Like Synoviocytes

Purpose: To investigate the effect of microRNA 498 (miR-498) on proliferation and inflammation of rheumatoid arthritis (RA) fibroblast-like synoviocytes (RA-FLSs) in rheumatoid arthritis (RA). Methods: MiR-498 level was evaluated in both RA synovial tissues and RA-FLSs using real-time polymerase chain reaction (PCR). MicroRNA-498 overexpression or knockdown was performed in RAFLSs. Proliferation, apoptosis, cell cycle and inflammation induced by miR-498 mimics or inhibitor were used to explore the function of miR-498 in RA. Results: Expression level of miR-498 was downregulated in both RA synovial tissues and RA- FLSs. MicroRNA-498 mimics decreased proliferation and arrested cell cycle, whereas miR-498 inhibitor caused the opposite effects in RA-FLSs. In addition, miR-498 mimics suppressed inflammation and promoted cell apoptosis, while miR-498 inhibitor promoted inflammation and inhibited cell apoptosis in RA-FLSs. Furthermore, the effect of miR-498 on the proliferation, inflammation and apoptosis of RAFLSs was mediated by its ability to target and downregulate JAK1. Conclusion: These results indicate that miR-498 inhibits the proliferation and inflammatory responses of RA-FLSs by targeting JAK1, thus revealing a new therapeutic target for RA treatment.

ATL

2014 ◽  
Vol 24 (2) ◽  
pp. 201-209 ◽  
Author(s):  
Cheng-Chang Chang ◽  
Rui-Lan Huang ◽  
Hui-Chen Wang ◽  
Yu-Ping Liao ◽  
Mu-Hsien Yu ◽  
...  
Keyword(s):  
Squamous Cell ◽  
Methylation Status ◽  
Squamous Cell Carcinomas ◽  
Odds Ratios ◽  
Chain Reaction ◽  
Bisulfite Pyrosequencing ◽  
Specific Polymerase Chain Reaction ◽  
The Status ◽  
Polymerase Chain ◽  
Normal Controls

ObjectiveThis study aimed to investigate the status of DNA methylation of 6 genes,LMX1A,NKX6-1,PAX1,PTPRR,SOX1, andZNF582, previously found from squamous cell carcinomas in adenocarcinomas (ACs) of the uterine cervix.MethodsWe assessed the methylation status of these genes in 40 ACs, cervical scrapings from 23 ACs, and 67 normal control cervices by real-time quantitative methylation-specific polymerase chain reaction. The results were validated by bisulfite pyrosequencing.ResultsThe methylation levels of all the 6 genes in the ACs were significantly higher than those in normal cervical tissues, especially forPAX1,PTPRR,SOX1, andZNF582. The odds ratios and 95% confidence intervals (CIs) of high methylation levels inPAX1,PTPRR,SOX1, andZNF582for the risk of developing an AC were 15.7 (95% CI, 7.0–40.6), 16.9 (95% CI, 7.6–43.0), 32.1 (95% CI, 12.1–124.3), and 25.4 (95% CI, 10.4–78.3), respectively (allP< 0.001). The methylation indices ofPAX1,PTPRR,SOX1, andZNF582recovered from scrapings of ACs were significantly higher than in normal controls. The odds ratios of these indices for the risk of developing an AC inPAX1,PTPRR,SOX1, andZNF582were 6.2 (95% CI, 2.6–15.4), 12.1(95% CI, 3.8–46.4), 6.2 (95% CI, 2.6–15.8), and 20.6 (95% CI, 6.9–77.5), respectively (allP< 0.001).ConclusionsCervical ACs carry aberrantly high methylation rates ofPAX1,PTPRR,SOX1, andZNF582—commonly methylated in squamous cell carcinomas—which might help for AC screening.

Gene-Specific Methylation: Potential Markers for Colorectal Cancer

2009 ◽  
Vol 24 (1) ◽  
pp. 57-62 ◽  
Author(s):  
Jyothi S. Prabhu ◽  
Aruna Korlimarla ◽  
Arindam Banerjee ◽  
Shivangi Wani ◽  
K Payal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Promoter Methylation ◽  
Methylation Status ◽  
Epigenetic Mechanism ◽  
Aberrant Methylation ◽  
Chain Reaction ◽  
Promoter Methylation Status ◽  
Polymerase Chain

Purpose Aberrant methylation of the promoter region is associated with silencing of many genes in neoplasia. CpG island methylation is an epigenetic mechanism for transcriptional silencing that occurs at various stages of colon tumorigenesis. In this study, we tested the promoter methylation and expression of seven genes from various pathways of DNA repair, apoptosis and inflammation, i.e., sFRP1, MLH1, RASSF1A, CDA, v-fgr, LYN-B, and TNFR10d. Method The genes were analyzed by quantitative polymerase chain reaction for the level of gene expression. The promoter methylation status of the genes was studied by methylation-specific polymerase chain reaction. Result The correlation of promoter methylation status with suppressed gene expression patterns suggested a potential role for the silencing these genes in colon cancer progression. Conclusion Promoter methylations of the studied genes could be explored as promising biomarkers for new diagnostic, prognostic and therapeutic targets of colorectal cancer.

Real-Time Methylation-Specific Polymerase Chain Reaction for MGMT Promoter Methylation Clinical Testing in Glioblastoma

2017 ◽  
Vol 148 (4) ◽  
pp. 296-307
Author(s):  
Cristiane M Ida ◽  
Malinda L Butz ◽  
Robert B Jenkins ◽  
Jann N Sarkaria ◽  
Gaspar J Kitange ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Promoter Methylation ◽  
Mgmt Promoter Methylation ◽  
Reference Laboratory ◽  
Clinical Testing ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Mgmt Promoter ◽  
Polymerase Chain

Abstract Objectives To develop and evaluate a real-time methylation-specific polymerase chain reaction (RT-MSP) MGMT assay, with a particular focus on small biopsies and indeterminate testing results. Methods We assessed formalin-fixed paraffin-embedded glioblastoma or gliosarcoma specimens (n = 641). A test-validation group (n = 51) with previously obtained reference laboratory (RL) results was used to determine performance characteristics of the RT-MSP assay. An indeterminate (equivocal) category was established for cases that could not be clearly classified as positive or negative. Results Overall agreement of RT-MSP and RL results was 91% (41/45 nonindeterminate cases). Discordant cases were tested by pyrosequencing, and results were most concordant with RT-MSP. Among cases with limited amounts of tissue (n = 7), six yielded valid results by RT-MSP (all negative); the single invalid result consisted of a stereotactic biopsy specimen obtained 14 years prior. A subset of indeterminate cases obtained during clinical testing (n = 18/575 [3%]) was also evaluated by pyrosequencing and showed a heterogeneous pattern of methylation across the eight interrogated CpG sites. Conclusions The RT-MSP assay that we developed in-house is a robust clinical detection method for the heterogeneous process of MGMT promoter methylation in glioblastoma.

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