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Epigenetically silenced miR-34b/c as a novel faecal-based screening marker for colorectal cancer.

Kalimutho M, Di Cecilia S, Del Vecchio Blanco G, Roviello F, Sileri P, Cretella M, Formosa A, Corso G, Marrelli D, Pallone F, Federici G, Bernardini S - Br. J. Cancer (2011)

Bottom Line: The miR-34b/c hypermethylation was found in 97.5% (79 out of 82) of primary colorectal tumours, P=0.0110.In addition, miR-148a was found to be hypermethylated in 65% (51 out of 78) of colorectal tumour tissues with no significant correlation to clinicopathological features.However, a trend with female gender and advanced age was found, P=0.083.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, University of Rome Tor Vergata, Rome, Italy. m.kalimutho@qub.ac.uk

ABSTRACT

Background: MicroRNAs are tiny non-coding small endogenous RNAs that regulate gene expression by translational repression, mRNA cleavage and mRNA inhibition. The aim of this study was to investigate the hypermethylation of miR-34b/c and miR-148a in colorectal cancer, and correlate this data to clinicopathological features. We also aimed to evaluate the hypermethylation of miR-34b/c in faeces specimens as a novel non-invasive faecal-DNA-based screening marker.

Methods: The 5-aza-2'-deoxycytidine treatment and methylation-specific PCR were carried out to detect the hypermethylation of miR-34b/c and miR-148a.

Results: The miR-34b/c hypermethylation was found in 97.5% (79 out of 82) of primary colorectal tumours, P=0.0110. In 75% (21 out of 28) of faecal specimens we found a hypermethylation of miR-34b/c while only in 16% (2 out of 12) of high-grade dysplasia. In addition, miR-148a was found to be hypermethylated in 65% (51 out of 78) of colorectal tumour tissues with no significant correlation to clinicopathological features. However, a trend with female gender and advanced age was found, P=0.083. We also observed a trend to lower survival rate in patients with miR-148a hypermethylation with 10-year survival probability: 48 vs 65%, P=0.561.

Conclusions: These findings show that aberrant hypermethylation of miR-34b/c could be an ideal class of early screening marker, whereas miR-148a could serve as a disease progression follow-up marker.

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Related in: MedlinePlus

Methylation-specific PCR (MSP) reactions for the miR-34b/c and miR-148a promoter region in tumour and faecal specimens derived from CRC and/or normal individual. (A and B) MSP analysis for miR-34b/c in matched CRC samples. (C) MSP analysis for miR-34b/c in faeces of CRC (upper panel), colonoscopy negative individuals (middle panel) and high-grade dysplasia (lower pane). (D) MSP analysis for miR-148a in matched CRC tissues. Lane UM and M corresponded to unmethylated and methylated reaction, respectively. Qiagen methylated and unmethylated control (CTRL) DNAs served as a reaction control for polymerase chain reaction. NTC=negative template control; POS=positive template control; P=patients; F-P1=samples derived from faeces of CRC and normal individuals.
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fig2: Methylation-specific PCR (MSP) reactions for the miR-34b/c and miR-148a promoter region in tumour and faecal specimens derived from CRC and/or normal individual. (A and B) MSP analysis for miR-34b/c in matched CRC samples. (C) MSP analysis for miR-34b/c in faeces of CRC (upper panel), colonoscopy negative individuals (middle panel) and high-grade dysplasia (lower pane). (D) MSP analysis for miR-148a in matched CRC tissues. Lane UM and M corresponded to unmethylated and methylated reaction, respectively. Qiagen methylated and unmethylated control (CTRL) DNAs served as a reaction control for polymerase chain reaction. NTC=negative template control; POS=positive template control; P=patients; F-P1=samples derived from faeces of CRC and normal individuals.

Mentions: We found that 97.5% (n=79 out of 81) of neoplastic samples showed promoter methylation of miR-34b/c (Figures 2A and B). In contrast, promoter methylation of miR-34b/c was only detected in 14.3% (n=6/42; P<0.001; Figures 2A and B, Table 4) of matched normal colonic tissues. No correlation were found for miR-34b/c methylation pattern with clinicopathological status except for one significant correlation for pTNM stage, P=0.0110 (Table 4). However, this result is conflicting as only two samples were not positive for stage II cancer.


Epigenetically silenced miR-34b/c as a novel faecal-based screening marker for colorectal cancer.

Kalimutho M, Di Cecilia S, Del Vecchio Blanco G, Roviello F, Sileri P, Cretella M, Formosa A, Corso G, Marrelli D, Pallone F, Federici G, Bernardini S - Br. J. Cancer (2011)

Methylation-specific PCR (MSP) reactions for the miR-34b/c and miR-148a promoter region in tumour and faecal specimens derived from CRC and/or normal individual. (A and B) MSP analysis for miR-34b/c in matched CRC samples. (C) MSP analysis for miR-34b/c in faeces of CRC (upper panel), colonoscopy negative individuals (middle panel) and high-grade dysplasia (lower pane). (D) MSP analysis for miR-148a in matched CRC tissues. Lane UM and M corresponded to unmethylated and methylated reaction, respectively. Qiagen methylated and unmethylated control (CTRL) DNAs served as a reaction control for polymerase chain reaction. NTC=negative template control; POS=positive template control; P=patients; F-P1=samples derived from faeces of CRC and normal individuals.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3111174&req=5

fig2: Methylation-specific PCR (MSP) reactions for the miR-34b/c and miR-148a promoter region in tumour and faecal specimens derived from CRC and/or normal individual. (A and B) MSP analysis for miR-34b/c in matched CRC samples. (C) MSP analysis for miR-34b/c in faeces of CRC (upper panel), colonoscopy negative individuals (middle panel) and high-grade dysplasia (lower pane). (D) MSP analysis for miR-148a in matched CRC tissues. Lane UM and M corresponded to unmethylated and methylated reaction, respectively. Qiagen methylated and unmethylated control (CTRL) DNAs served as a reaction control for polymerase chain reaction. NTC=negative template control; POS=positive template control; P=patients; F-P1=samples derived from faeces of CRC and normal individuals.
Mentions: We found that 97.5% (n=79 out of 81) of neoplastic samples showed promoter methylation of miR-34b/c (Figures 2A and B). In contrast, promoter methylation of miR-34b/c was only detected in 14.3% (n=6/42; P<0.001; Figures 2A and B, Table 4) of matched normal colonic tissues. No correlation were found for miR-34b/c methylation pattern with clinicopathological status except for one significant correlation for pTNM stage, P=0.0110 (Table 4). However, this result is conflicting as only two samples were not positive for stage II cancer.

Bottom Line: The miR-34b/c hypermethylation was found in 97.5% (79 out of 82) of primary colorectal tumours, P=0.0110.In addition, miR-148a was found to be hypermethylated in 65% (51 out of 78) of colorectal tumour tissues with no significant correlation to clinicopathological features.However, a trend with female gender and advanced age was found, P=0.083.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine, University of Rome Tor Vergata, Rome, Italy. m.kalimutho@qub.ac.uk

ABSTRACT

Background: MicroRNAs are tiny non-coding small endogenous RNAs that regulate gene expression by translational repression, mRNA cleavage and mRNA inhibition. The aim of this study was to investigate the hypermethylation of miR-34b/c and miR-148a in colorectal cancer, and correlate this data to clinicopathological features. We also aimed to evaluate the hypermethylation of miR-34b/c in faeces specimens as a novel non-invasive faecal-DNA-based screening marker.

Methods: The 5-aza-2'-deoxycytidine treatment and methylation-specific PCR were carried out to detect the hypermethylation of miR-34b/c and miR-148a.

Results: The miR-34b/c hypermethylation was found in 97.5% (79 out of 82) of primary colorectal tumours, P=0.0110. In 75% (21 out of 28) of faecal specimens we found a hypermethylation of miR-34b/c while only in 16% (2 out of 12) of high-grade dysplasia. In addition, miR-148a was found to be hypermethylated in 65% (51 out of 78) of colorectal tumour tissues with no significant correlation to clinicopathological features. However, a trend with female gender and advanced age was found, P=0.083. We also observed a trend to lower survival rate in patients with miR-148a hypermethylation with 10-year survival probability: 48 vs 65%, P=0.561.

Conclusions: These findings show that aberrant hypermethylation of miR-34b/c could be an ideal class of early screening marker, whereas miR-148a could serve as a disease progression follow-up marker.

Show MeSH
Related in: MedlinePlus