Methylation-Sensitive Melt Curve Analysis of the Reprimo Gene Methylation in Gastric Cancer

Wang, Hanze; Zheng, Yu; Lai, Junzhong; Luo, Qianping; Ke, Huican; Chen, Qi

doi:10.1371/journal.pone.0168635

Cited by 14 publications

(14 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We used bisulfite sequencing and MS-MCA to evaluate the methylation status of the RPRM promoter in the tumor and para-cancerous tissues, and plasma samples of gastric cancer patients and normal healthy individuals. Consistent with our methylation-sensitive melt curve analyses [ 40 ], the RPRM promoter methylation was detected in all of the tumor tissues of gastric cancer patients. However, for some para-cancerous tissues, RPRM was also methylated, which suggests that the cellular fate of the para-cancerous tissues may have been changed or transformed, which is consistent with previous observations [ 4 ].…”

Section: Discussionsupporting

confidence: 87%

The relationship between DNA methylation andReprimogene expression in gastric cancer cells

et al. 2017

Self Cite

View full text Add to dashboard Cite

Reprimo (RPRM) is a tumor suppressor involved in the development of a number of malignant tumors including gastric cancer which is highly related to its gene hypermethylation. However, the regulation of RPRM gene expression by DNA methylation in gastric cancer is not well understood. We examined the RPRM gene methylation in gastric cancer tissues or plasma samples by bisulfite sequencing, and investigated the relationship between DNA methylation and the RPRM gene expression by quantitative reverse transcription-PCR and Western blotting. We found that the RPRM gene promoter region is hypermethylated in gastric cancer tissues (75%, 45/60), plasma samples (86.3%, 44/51) and various cancer cell lines (75%, 3/4), which is correlated with the decrease of RPRM gene expression. The hypermethylation-induced RPRM reduction can be recovered by treating with zebularine, a demethylating agent, and by inhibition of the DNA methyltransferases via RNA interference and CRISPR/Cas9-mediated gene knockout. In addition, we generated RPRM gene-knockout cells and studied the effects of the RPRM deficiency on tumor formation by inoculating these cells in mice. The data show that the loss of RPRM can promote tumorigenesis. These data suggest that the RPRM expression is inhibited by DNA methyltransferases and the RPRM normal function can be restored by treating with DNA methylation inhibitors. The study provides important information regarding the role of RPRM and its methylation related to gastric cancer development.

show abstract

Section: Discussionsupporting

confidence: 87%

The relationship between DNA methylation andReprimogene expression in gastric cancer cells

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition, Ooki et al indicated that clinical assessment of RPRM methylation may also serve as a predictive marker for response to chemotherapy consisting of cisplatin and fluoropyrimidines, and as a marker of tumor aggressiveness [ 18 ]. Wang et al [ 38 ] recently published an article where they developed a semi-quantitative method based on MS-MCA for detecting DNA methylation of the RPRM gene in human plasma or serum samples to help in the diagnosis/prognosis of gastric cancer. Recently, an interesting study by Garcia-Bloj et al [ 39 ] developed a combinatorial strategy for the reactivation of tumor suppressor genes, including RPRM , using CRISPR/Cas9 VP64 with synergistic activation mediators, which led to phenotypic reprogramming in AGS gastric cancer cells.…”

Section: Discussionmentioning

confidence: 99%

Reprimo, a Potential p53-Dependent Tumor Suppressor Gene, Is Frequently Hypermethylated in Estrogen Receptor α-Positive Breast Cancer

Buchegger

Riquelme

Viscarra

et al. 2017

IJMS

View full text Add to dashboard Cite

Aberrant DNA methylation is a hallmark of many cancers. Currently, there are four intrinsic molecular subtypes in breast cancer (BC): Luminal A, B, Her2-positive, and triple negative (TNBC). Recently, The Cancer Genome Atlas (TCGA) project has revealed that Luminal subtypes have higher levels of genome-wide methylation that may be a result of Estrogen/Estrogen receptor α (E2/ERα) signaling pathway activation. In this study, we analyze promoter CpG-island (CGIs) of the Reprimo (RPRM) gene in breast cancers (n = 77), cell lines (n = 38), and normal breast tissue (n = 10) using a MBDCap-seq database. Then, a validation cohort (n = 26) was used to confirm the results found in the MBDCap-seq platform. A differential methylation pattern was found between BC and cell lines compared to normal breast tissue. In BC, a higher DNA methylation was observed in tissues that were ERα-positive than in ERα-negative ones; more precisely, subtypes Luminal A compared to TNBC. Also, significant reverse correlation was observed between DNA methylation and RPRM mRNA expression in BC. Our data suggest that ERα expression in BC may affect the DNA methylation of CGIs in the RPRM gene. This approach suggests that DNA methylation status in CGIs of some tumor suppressor genes could be driven by E2 availability, subsequently inducing the activation of the ERα pathway.

show abstract

“…Some studies reported that promoter methylation of the p16 , CDH1 , RUNX3 , MLH1 , RASSF1A , p15 , APC , DAPK , GSTP1 , Reprimo , and MGMT was frequent in the blood in patients with GC [ 34 – 38 ]. However, the results of the frequencies of promoter methylation of these 11 tumor-related genes are conflicting and different in blood samples of GC patients and non-tumor controls.…”

Section: Discussionmentioning

confidence: 99%

“…GSTP1 promoter methylation had a frequency of < 20% in blood samples of GC patients [ 29 , 38 ]. Reprimo promoter methylation had a high rate in blood samples of patients with GC (> 60%) [ 22 , 34 , 46 ]. Leung 2005 et al showed no correlation between MGMT promoter methylation and GC [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

Promoter methylation of tumor-related genes as a potential biomarker using blood samples for gastric cancer detection

Wen

Zheng

et al. 2017

Oncotarget

View full text Add to dashboard Cite

Gene promoter methylation has been reported in gastric cancer (GC). However, the potential applications of blood-based gene promoter methylation as a noninvasive biomarker for GC detection remain to be evaluated. Hence, we performed this analysis to determine whether promoter methylation of 11 tumor-related genes could become a promising biomarker in blood samples in GC. We found that the cyclin-dependent kinase inhibitor 2A (p16), E-cadherin (CDH1), runt-related transcription factor 3 (RUNX3), human mutL homolog 1 (MLH1), RAS association domain family protein 1A (RASSF1A), cyclin-dependent kinase inhibitor 2B (p15), adenomatous polyposis coli (APC), Glutathione S-transferase P1 (GSTP1), TP53 dependent G2 arrest mediator candidate (Reprimo), and O6-methylguanine-DNAmethyl-transferase (MGMT) promoter methylation was notably higher in blood samples of patients with GC compared with non-tumor controls. While death-associated protein kinase (DAPK) promoter methylation was not correlated with GC. Further analyses demonstrated that RUNX3, RASSF1A and Reprimo promoter methylation had a good diagnostic capacity in blood samples of GC versus non-tumor controls (RUNX3: sensitivity = 63.2% and specificity = 97.5%, RASSF1A: sensitivity = 61.5% and specificity = 96.3%, Reprimo: sensitivity = 82.0% and specificity = 89.0%). Our findings indicate that promoter methylation of the RUNX3, RASSF1A and Reprimo genes could be powerful and potential noninvasive biomarkers for the detection and diagnosis of GC in blood samples in clinical practices, especially Reprimo gene. Further well-designed (multi-center) and prospective clinical studies with large populations are needed to confirm these findings in the future.

show abstract

Methylation-Sensitive Melt Curve Analysis of the Reprimo Gene Methylation in Gastric Cancer

Cited by 14 publications

References 19 publications

The relationship between DNA methylation andReprimogene expression in gastric cancer cells

The relationship between DNA methylation andReprimogene expression in gastric cancer cells

Reprimo, a Potential p53-Dependent Tumor Suppressor Gene, Is Frequently Hypermethylated in Estrogen Receptor α-Positive Breast Cancer

Promoter methylation of tumor-related genes as a potential biomarker using blood samples for gastric cancer detection

Contact Info

Product

Resources

About