Methylation Status of hMLH1 Gene in Colorectal Cancer Patients in Ethenic Population of Kashmir Valley

384 Background: Identification of blood-based biomarkers for cancer screening is essential in order to develop novel and minimally invasive methods for colorectal cancer screening. Our lab has successfully applied a novel nanotechnology that allows us to detect and amplify a single tumor DNA fragment in a plasma sample. This DNA is tested for methylation of several genes including TFPI2 which has shown to be highly sensitive and specific for the detection colorectal cancer in stool. Methods: Whole blood was obtained from 18 colorectal cancer patients and plasma was isolated. Plasma was processed using Methylation On Beads nanotechnology (MOB) and bisulfate treated. Methylation status was determined via quantitative PCR method. Results: Two genes, TFPI2 and IGFBP3, were detected with a high sensitivity. TFPI2, demonstrated a methylation frequency of 94.4%, which is concordant with the TFPI2 methylation frequency of 99% in primary colorectal cancer tissues. IGFBP3 showed the methylation frequency of 61.1%, which corresponds with the methylation frequency of 52% in retrospective colorectal cancer tissues in previous studies. Quantification using standard curves indicated a single copy level of DNA found in plasma. Conclusions: Blood-based screening is challenging due to extremely low quantities of circulating DNA in blood. Utilizing a novel nanotechnology that detects DNA at a single copy level, the methylation changes in colorectal cancer were successfully detected in plasmas at similar frequencies as in tissue samples. This study has demonstrated the feasablility and applicability to blood-based screening. Future studies will focus on improving the sensitivity and determining the specificity of this method.

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Differential survival trends of stage II colorectal cancer patients relate to promoter methylation status of PCDH10, SPARC, and UCHL1

Modern Pathology ◽

10.1038/modpathol.2013.204 ◽

2013 ◽

Vol 27 (6) ◽

pp. 906-915 ◽

Cited By ~ 16

Author(s):

Ellen Heitzer ◽

Monika Artl ◽

Martin Filipits ◽

Margit Resel ◽

Ricarda Graf ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Promoter Methylation ◽

Methylation Status ◽

Stage Ii ◽

Differential Survival ◽

Promoter Methylation Status ◽

Stage Ii Colorectal Cancer ◽

Colorectal Cancer Patients

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Methylation status of DLEC1 promoter in colorectal cancer patients and its clinical relevance

Academic Journal of Second Military Medical University ◽

10.3724/sp.j.1008.2010.00842 ◽

2010 ◽

Vol 30 (8) ◽

pp. 842-845

Author(s):

Xiao-bing YE ◽

You-wei ZHANG ◽

Long-bang CHEN

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Clinical Relevance ◽

Methylation Status ◽

Colorectal Cancer Patients

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Distinct epigenetic features of tumor-reactive CD8+ T cells in colorectal cancer patients revealed by genome-wide DNA methylation analysis

Genome Biology ◽

10.1186/s13059-019-1921-y ◽

2019 ◽

Vol 21 (1) ◽

Cited By ~ 4

Author(s):

Rui Yang ◽

Sijin Cheng ◽

Nan Luo ◽

Ranran Gao ◽

Kezhuo Yu ◽

...

Keyword(s):

Colorectal Cancer ◽

Dna Methylation ◽

T Cells ◽

T Cell ◽

Cancer Patients ◽

Cd8 T Cells ◽

Methylation Status ◽

Effector Memory ◽

Binding Motif ◽

Colorectal Cancer Patients

Abstract Background Tumor-reactive CD8+ tumor-infiltrating lymphocytes (TILs) represent a subtype of T cells that can recognize and destroy tumor specifically. Understanding the regulatory mechanism of tumor-reactive CD8+ T cells has important therapeutic implications. Yet the DNA methylation status of this T cell subtype has not been elucidated. Results In this study, we segregate tumor-reactive and bystander CD8+ TILs, as well as naïve and effector memory CD8+ T cell subtypes as controls from colorectal cancer patients, to compare their transcriptome and methylome characteristics. Transcriptome profiling confirms previous conclusions that tumor-reactive TILs have an exhausted tissue-resident memory signature. Whole-genome methylation profiling identifies a distinct methylome pattern of tumor-reactive CD8+ T cells, with tumor-reactive markers CD39 and CD103 being specifically demethylated. In addition, dynamic changes are observed during the transition of naïve T cells into tumor-reactive CD8+ T cells. Transcription factor binding motif enrichment analysis identifies several immune-related transcription factors, including three exhaustion-related genes (NR4A1, BATF, and EGR2) and VDR, which potentially play an important regulatory role in tumor-reactive CD8+ T cells. Conclusion Our study supports the involvement of DNA methylation in shaping tumor-reactive and bystander CD8+ TILs, and provides a valuable resource for the development of novel DNA methylation markers and future therapeutics.

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The Differential DNA Hypermethylation Patterns of microRNA-137 and microRNA-342 Locus in Early Colorectal Lesions and Tumours

Biomolecules ◽

10.3390/biom9100519 ◽

2019 ◽

Vol 9 (10) ◽

pp. 519

Author(s):

Elham Kashani ◽

Mahrooyeh Hadizadeh ◽

Vahid Chaleshi ◽

Reza Mirfakhraie ◽

Chris Young ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Methylation Status ◽

Dna Hypermethylation ◽

Normal Colon Mucosa ◽

Normal Tissues ◽

Diagnosis And Prognosis ◽

Adjacent Mucosa ◽

Prediction And Prevention ◽

Colorectal Cancer Patients

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, representing 13% of all cancers. The role of epigenetics in cancer diagnosis and prognosis is well established. MicroRNAs in particular influence numerous cancer associated processes including apoptosis, proliferation, differentiation, cell-cycle controls, migration/invasion and metabolism. MiRNAs-137 and 342 are exon- and intron-embedded, respectively, acting as tumour-suppressive microRNA via hypermethylation events. Levels of miRNAs 137 and 342 have been investigated here as potential prognostic markers for colorectal cancer patients. The methylation status of miRNA-137 and miRNA-342 was evaluated using methylation-specific (MSP) polymerase chain reaction (PCR) on freshly frozen tissue derived from 51 polyps, 8 tumours and 14 normal colon mucosa specimens. Methylation status of miRNA-137 and miRNA-342 was significantly higher in tumour lesions compared to normal adjacent mucosa. Surprisingly, the methylation frequency of miR-342 (76.3%) among colorectal cancer patients was significantly higher compared to miR-137 (18.6%). Furthermore, normal tissues, adjacent to the lesions (N-Cs), displayed no observable methylation for miRNA-137, whereas 27.2% of these N-Cs showed miRNA-342 hypermethylation. MiRNA-137 hypermethylation was significantly higher in male patients and miR-342 hypermethylation correlated with patient age. Methylation status of miRNA-137 and miRNA-342 has both diagnostic and prognostic value in CRC prediction and prevention.

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Two novel mutations in hMLH1 gene in Iranian hereditary non-polyposis colorectal cancer patients

Familial Cancer ◽

10.1007/s10689-011-9478-2 ◽

2011 ◽

Vol 11 (1) ◽

pp. 13-17 ◽

Cited By ~ 3

Author(s):

Somayeh Shahmoradi ◽

Ali Bidmeshkipour ◽

Ahmad Salamian ◽

Mohammad Hasan Emami ◽

Zahra Kazemi ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Patients ◽

Novel Mutations ◽

Colorectal Cancer Patients ◽

Hmlh1 Gene

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Could CYP24A1 promoter methylation status affect the gene expression in the colorectal cancer patients?

Meta Gene ◽

10.1016/j.mgene.2020.100656 ◽

2020 ◽

Vol 24 ◽

pp. 100656

Author(s):

Hossein Sadeghi ◽

Ehsan Nazemalhosseini-Mojarad ◽

Vahid Reza Yassaee ◽

Sanaz Savabkar ◽

Majid Ghasemian ◽

...

Keyword(s):

Gene Expression ◽

Colorectal Cancer ◽

Cancer Patients ◽

Promoter Methylation ◽

Methylation Status ◽

Promoter Methylation Status ◽

Colorectal Cancer Patients

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Methylations of NEUROG1, p16, and MLH1 and recurrence following adjuvant FOLFOX in colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2012.30.15_suppl.3624 ◽

2012 ◽

Vol 30 (15_suppl) ◽

pp. 3624-3624

Author(s):

Hyun-Jung Lee ◽

Sae-Won Han ◽

Jeong Mo Bae ◽

Nam-Yun Cho ◽

Kyung-Hun Lee ◽

...

Keyword(s):

Colorectal Cancer ◽

Treatment Outcome ◽

Cancer Patients ◽

Cpg Island ◽

Methylation Status ◽

Disease Free Survival ◽

Sporadic Colorectal Cancer ◽

Prognostic Impact ◽

Surgical Specimen ◽

Colorectal Cancer Patients

3624 Background: CpG island methylator phenotype (CIMP) is characterized by concurrent methylation of multiple CpG islands in tumor DNA, which can inactivate tumor suppressor genes or promote carcinogenesis. The prognostic impact of CIMP on treatment outcome of colon cancer patients receiving adjuvant 5-fluorouracil/leucovorin/oxaliplatin (FOLFOX) is unclear. We investigated CIMP markers in colorectal cancer patients treated with adjuvant FOLFOX. Methods: Sporadic colorectal cancer patients treated with curative resection followed by adjuvant FOLFOX were included. DNA was extracted from formalin-fixed paraffin-embedded surgical specimen. 8 CpG island loci (CACNA1G, CRABP1, IGF2, MLH1, NEUROG1, P16, RUNX3 and SOCS1) were examined using MethyLight analysis. Disease-free survival (DFS) was evaluated according to each methylation loci. Results: A total of 322 patients were included. Methylation at 1 or more loci was observed in 150 patients (46.6%) and 6 or more loci in 15 (4.7%). During a median follow-up duration of 39.7 months, 55 recurrences were observed. Three year DFS in the patient cohort was 84%. CRABP1 (23.9%) was the most frequently methylated loci, followed by p16 (22.7%) and NEUROG1 (20.8%). Patients having methylation at NEUROG1 (3 year DFS 78% in (+) vs. 86% in (-), p = 0.014) and p16 (3 year DFS 78% in (+) vs. 86% in (-), p = 0.12) had worse DFS, whereas methylation at MLH1 had better DFS (3 year DFS 100% in (+) vs. 86% in (-), p = 0.19). In a combined analysis, patients with MLH1(-)/NEUROG1(+)/p16(+) had worst treatment outcome compared to MLH1(-)/NEUROG1(+) or p16(+), MLH1(-)/ NEUROG1(-) /p16(-), and MLH1(+) (3 year DFS 62%, 82%, 87%, and 100%, respectively; p = 0.002). In multivariate analysis, NEUROG1(+)/p16(+) was associated with significantly higher recurrence compared with other patients (adjusted hazard ratio (HR) 2.15 (95% confidence interval (CI) 1.08 - 4.27, p = 0.029). Conclusions: Methylation status of NEUROG1, p16, and MLH1 is associated with recurrence following adjuvant FOLFOX in stage II/III colorectal cancer. Further validation and translational studies to improve treatment outcome in the subset of patients are warranted in the future.

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