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 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.

Microsporidial infections due toEncephalitozoon intestinalisin non-HIV-infected patients with chronic diarrhoea

2011 ◽  
Vol 140 (10) ◽  
pp. 1773-1779 ◽  
Author(s):  
J. YAKOOB ◽  
Z. ABBAS ◽  
M. ASIM BEG ◽  
W. JAFRI ◽  
S. NAZ ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Stool Examination ◽  
Chronic Diarrhoea ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Specific Primers ◽  
Specific Pcr ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Species Specific

SUMMARYWe determined the prevalence of microsporidiaEnterocytozoon(Ent.)bieneusiandEncephalitozoon(E.)intestinalisinfection in patients with chronic diarrhoea and hepatocellular carcinoma (HCC). A total of 330 stool samples were examined from 171 (52%) patients with chronic diarrhoea, 18 (5%) with HCC while 141 (43%) were controls. Stool microscopy, polymerase chain reaction (PCR) with species-specific primers forEnt. bieneusiandE. intestinalisand sequencing were carried out. Microsporidia were found by trichrome staining in 11/330 (3%) andE. intestinalisby PCR in 13/330 (4%) whileEnt. bieneusiwas not detected. PCR forE. intestinaliswas positive in 8/171 (5%) stool samples from patients with chronic diarrhoea, 2/141 (1·4%) samples from healthy controls and in 3/18 (17%) samples from patients with HCC. In the chronic diarrhoea group,E. intestinaliswas positive in 4/171 (2·3%) (P=0·69) stool samples compared to 2/18 (11%) (P=0·06) in the HCC group and 2/141 (1·4%) from healthy controls.E. intestinalisinfection was significantly associated with chronic diarrhoea and HCC in these patients who were negative for HIV. Stool examination with trichrome or species-specific PCR for microsporidia may help establish the cause of chronic diarrhoea.

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.

scholarly journals Entamoeba species associated with chronic diarrhoea in Pakistan

2011 ◽  
Vol 140 (2) ◽  
pp. 323-328 ◽  
Author(s):  
J. YAKOOB ◽  
Z. ABBAS ◽  
M. A. BEG ◽  
S. NAZ ◽  
R. KHAN ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Irritable Bowel Syndrome ◽  
Abdominal Pain ◽  
Chronic Diarrhoea ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Stool Samples ◽  
Polymerase Chain ◽  
Irritable Bowel

SUMMARYWe determined the prevalence of Entamoeba (E.) histolytica, E. dispar and E. moshkovskii in patients with chronic diarrhoea associated with abdominal pain or discomfort mimicking irritable bowel syndrome. Stool samples were collected from 161 patients with chronic diarrhoea and from 157 healthy controls. Stool microscopy with modified trichrome stain, culture and polymerase chain reaction (PCR) for Entamoeba spp. differentiation was performed. Microscopy demonstrated Entamoeba cysts in 44% (57/129) of patients with diarrhoea compared to 29% (44/151) of controls (P=0·009). In patients with diarrhoea, PCR for E. histolytica was positive in 9% (11/129) (P=0·008), E. dispar in 19% (24/129) (P=0·117) and E. moshkovskii in 19% (24/129) (P<0·001). E. histolytica and E. moshkovskii were significantly associated with diarrhoea while E. dispar was found equally in both groups.

scholarly journals Molecular Detection of Prostate Cancer by Methylation-Specific Polymerase Chain Reaction from Urine Specimens

2013 ◽  
Vol 32 (3) ◽  
pp. 233-237 ◽  
Author(s):  
Raluca Dumache ◽  
Sorina Popescu ◽  
Radu Minciu ◽  
Serban Negru ◽  
Maria Puiu
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Methylation Status ◽  
Promoter Hypermethylation ◽  
Chain Reaction ◽  
Noninvasive Methods ◽  
Specific Polymerase Chain Reaction ◽  
Gstp1 Gene ◽  
Polymerase Chain ◽  
Urine Specimens

Summary Background: Prostate cancer (PCa) represents the second most prevalent malignancy among males, which is characterized by a high mortality rate. The aim of our study was to evaluate the methylation status of glutathione S-transferase P1 (GSTP1) in urine specimens from males with PCa and benign prostatic hyperplasia (BPH) and its usefulness in distinguishing between males with PCa and BPH by noninvasive methods. Methods: Voided urine specimens were collected from 65 patients with PCa and 45 patients with BPH. Genomic DNA was isolated and subjected to bisulfite modification. Methylation status of the GSTP1 gene was determined by conventional methylation-specific polymerase chain reaction (MSP) analysis. Results: Promoter hypermethylation of the GSTP1 gene in voided urine samples was found in 63 of 65 (97%) males with PCa and in 5 of 45 (11%) males with BPH. The sensitivity and specificity of GSTP1 in discriminating between PCa and BPH males were 98% and 89%, respectively. Conclusions: Gene analysis of GSTP1 using conventional MSP in urine specimens can be used as a noninvasive biomarker to distinguish between men with malignant and benign prostatic diseases.

scholarly journals Hypomethylation of the DAZ3 promoter in idiopathic asthenospermia: a screening tool for liquid biopsy

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Shichang Zhang ◽  
Li Xu ◽  
Mengyao Yu ◽  
Jiexin Zhang
Keyword(s):  
Polymerase Chain Reaction ◽  
Blood Cells ◽  
Methylation Status ◽  
White Blood Cells ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Deleted In Azoospermia ◽  
Polymerase Chain ◽  
Peripheral White Blood Cells ◽  
Methylation Ratio

Abstract Given the role of the deleted in azoospermia gene in male infertility, whether the somatic deleted in azoospermia methylation status is associated with idiopathic asthenospermia should be determined. To investigate the methylation levels of the deleted in azoospermia promoter in peripheral white blood cells from idiopathic asthenospermia patients relative to those in normozoospermia controls, 61 ethylene diamine tetraacetic acid anticoagulant blood samples were drawn from all participants for DNA isolation. The deleted in azoospermia promoter methylation ratio was detected by MassARRAY-based methylation quantification and confirmed by quantitative methylation-specific polymerase chain reaction. A MassARRAY-based methylation analysis showed that the deleted in azoospermia 3 promoter (0 to − 2 kbp) was significantly hypomethylated in peripheral white blood cells from idiopathic asthenospermia males, specifically one CpG site (− 246 to − 247). Quantitative methylation-specific polymerase chain reaction data further confirmed that the methylation level of the deleted in azoospermia 3 promoter region in idiopathic asthenospermia patients was significantly lower than that in normozoospermia males. The area under the receiver operating characteristic curve determined by quantitative methylation-specific polymerase chain reaction was 0.737 (95% confidence interval: 0.552 to 0.924), with a sensitivity of 53.9% and a specificity of 88.2% at a cut-off level of 74.7%. Therefore, our results suggested that methylation ratio detection of the deleted in azoospermia 3 promoter region by real-time polymerase chain reaction assay is a promising and feasible tool for liquid biopsy in the clinical laboratories. The methylation status of other reported infertility-related genes should also be investigated in peripheral white blood cells.

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