Associations Between TFPI-2 Methylation and Poor Prognosis in Glioblastomas Paulina Vaitkienė, Daina Skiriutė, Kęstutis Skauminas

Background and Objective. The epigenetic silencing of tumor suppressor genes plays an important role in gliomagenesis. Recently, tissue factor pathway inhibitor 2 (TFPI-2) has been suggested as a tumor suppressor gene involved in tumorigenesis and metastasis in some cancers. However, to date, little is known about the methylation status of TFPI-2 gene in glioblastoma tissues. In this study, we aimed to investigate the methylation status of TFPI-2 promoter and its associations with patient prognosis in glioblastoma. Material and Methods. The methylation status of TFPI-2 was investigated by methylationspecific polymerase chain reaction in 99 glioblastoma patients. The associations between patients’ clinical variables and overall survival time were assessed. Results. TFPI-2 was aberrantly methylated in 22.2% (22/99) of glioblastoma tumors, but was not methylated in normal brain samples. The survival of patients with glioblastoma differed significantly between the methylated and unmethylated TFPI-2 groups (P=0.047). The 2-year survival among patients carrying methylated TFPI-2 tumors was significantly lower compared with that of patients with unmethylated TFPI-2 (27% versus 4.7%, P=0.037). Conclusions. The present work demonstrated that the epigenetic inactivation of TFPI-2 by promoter hypermethylation was a frequent and tumor-specific event in glioblastoma, and TFPI-2 promoter methylation might be considered as a prognostic marker in glioblastoma.

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Tissue factor pathway inhibitor-2 as a frequently silenced tumor suppressor gene in hepatocellular carcinoma

Hepatology ◽

10.1002/hep.21578 ◽

2007 ◽

Vol 45 (5) ◽

pp. 1129-1138 ◽

Cited By ~ 68

Author(s):

Chun-Ming Wong ◽

Yeung-Lam Ng ◽

Joyce Man-Fong Lee ◽

Carmen Chak-Lui Wong ◽

Oi-Fung Cheung ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Tumor Suppressor ◽

Tissue Factor ◽

Tumor Suppressor Gene ◽

Tissue Factor Pathway Inhibitor ◽

Suppressor Gene ◽

Tissue Factor Pathway

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Promoter Methylation of Multiple Genes in Germinal Center Hyperplasia and Lymphomas of Germinal Center Origin

Blood ◽

10.1182/blood.v112.11.4487.4487 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4487-4487

Author(s):

Jose M Paz-Carreira ◽

Raquel Losada ◽

Arantxa Garcia-Rivero ◽

Augusto Alvarez ◽

Fernando Bal ◽

...

Keyword(s):

Dna Methylation ◽

Tumor Suppressor ◽

B Cell ◽

Promoter Methylation ◽

Germinal Center ◽

Tumor Suppressor Genes ◽

Methylation Status ◽

Promoter Hypermethylation ◽

Epigenetic Mechanism ◽

Suppressor Genes

Abstract INTRODUCTION. Germinal centers (GC) are unique sites in peripheral lymphoid tissue where clonal selection of B cells takes place. This occurs as a response to stimulation by various antigens originating, sometimes, follicular hyperplasia (FH). GC have been known as a major source of B-cell lymphomas including follicular (FL) and diffuse large cell (DLCL). DNA methylation of tumor-suppressor genes is a mechanism of gene silencing involved in the pathogenesis of FL and DLCL. Much less is known about the role of methylation in FH. We determined the methylation status of 6 tumor-suppressor genes in 43 patients with FH, 18 patients with FL and 49 patients with DLCL in order to see the differential implication of this epigenetic mechanism in the pathological and the physiological development of GC. MATERIAL AND METHODS. Genomic DNA extracted from paraffin-embedded samples of 43 FH, 18 FL and 49 DLCL after being treated with EZ DNA-Methylation Kit (Zymo Research) with the manufacturer’s instructions, were analyzed by methylation-specific polymerase chain-reaction to determine promoter hypermethylation of DAP-k, SHP1, Rarβ, p14, SHP1, MGMT and PDRM1. All samples were obtained mostly from lymph nodes and tonsils. Diagnosis was based on morphology and immunohistochemistry analysis. All cases were matched for age, sex and ethnic origin. RESULTS: DAP-k promoter methylation occurred with higher frequency in FL(89%) than in DLCL(78%) and FH(40%). SHP1 was methylated in 61% of FL, 58% of FH and 23% of DLCL. RARb was methylated in 67% of FL patients, 30% of DLCL and only 12% of FH. Eight (44%) FL, seventeen (35%) DLCL and four (10%) BFH patients showed MGMT methylation. Promoter hypermethylation of p14 was detected only in 5 (12%) FH, 2 (4%) DLCL and none FL patients. Methylation of PRMD1 was present only in 1 (6%) FL, 2 (6%) DLCL and 1 (4%) FH samples. CONCLUSIONS. Inactivacion of DAP-K and SHP1 is present in B-cell malignancies, DLCL and FL, and BFH. Therefore, it may represent a physiologic event conferring a temporal survival advantage necessary for a GC hyperplastic response. Inactivation of the retinoic acid response through the methylation of Rarâ is significantly more frequent in lymphomas than in FH. As reported in other tumors methylation of MGMT is more frequent in lymphomas than in FH. With our data methylation of Cyclin dependent kinase inhibitors p14 is not a differential pathogenic event in lymphomas of GC origin, in fact it is more frequent in FH. Promoter Methylation of PDRM1 is not the mechanism involved in lymphomagenesis in FL and DLCL, the two FH positive deserve further follow-up to determine its significance.

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Detecting Aberrantly Methylated Genes in HNSCC Saliva

Otolaryngology ◽

10.1016/j.otohns.2008.05.493 ◽

2008 ◽

Vol 139 (2_suppl) ◽

pp. P89-P90

Author(s):

Seema Sethi ◽

Kang-Mei Chen ◽

Michael S Benninger ◽

Maria J Worsham

Keyword(s):

Tumor Suppressor ◽

Tumor Suppressor Genes ◽

Methylation Status ◽

Simultaneous Detection ◽

Promoter Hypermethylation ◽

Early Event ◽

Gene Probe ◽

Aberrant Methylation ◽

Suppressor Genes ◽

Normal Controls

Problem Aberrant promoter hypermethylation of multiple tumor suppressor genes is an early event in carcinogenesis of HNSCC. Saliva is a non-invasive sample which has immense potential for use in molecular screening and early diagnosis of head and neck cancers due to ease of collection, patient compliance allowing repeatabililty. We evaluated saliva DNA for simultaneous detection of promoter hypermethylation in multiple genes in HNSCC cases and compared them with normal controls. Methods Saliva was collected in a prospective cohort of 37 study subjects: 27 HNSCC patients and 10 normal controls. Saliva DNA was interrogated for aberrant methylation status using a multicandidate gene probe panel. 35 unique genes related to HNSCC were simultaneously examined of which 22 were tumor suppressor genes. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) assay was used. Results Of the 22 tumor suppressor genes examined simultaneously in the saliva DNA of the study subjects, four HNSCC cases showed aberrant promoter hypermethylation in one or more genes. Aberrantly methylated genes included TIMP3, APC, MLH1, RARB and CDKN1B, CDKN2B, GSTP1, CD44, and ESR. aberrant methylation of CDKN2B was observed in 3/4 cases and GSTP1 in 2/4 cases. Only one of the 10 controls had aberrant methylation of CDKN2B. Conclusion In this exploratory study, MS-MLPA identified multiple aberrantly methylated genes simultaneously in the same assay run. The most important advantage is its utility as a high throughput multi-gene screening approach requiring very small amounts of DNA, unlike more laborious single gene methods such as MSP. Significance Epigenetic signatures from MS-MLPA profiling, upon subsequent validation as diagnostic or prognostic biomarkers, can become reduced to a more definitive candidate gene panel of only a few key genes for early detection and screening of HNSCC. Support HFHS A10236; NIH R01 DE15990.

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Prognostic Significance of Promoter Methylation of Multiple Genes in Germinal Center Hyperplasia (GCH) and Lymphomas of Germinal Center Origin.

Blood ◽

10.1182/blood.v114.22.3950.3950 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3950-3950

Author(s):

Jose M Paz-Carreira ◽

Arantxa Garcia-Rivero ◽

Raquel Losada ◽

Jose C Mendez ◽

Manuel Albors ◽

...

Keyword(s):

Overall Survival ◽

Tumor Suppressor ◽

Promoter Methylation ◽

Germinal Center ◽

Tumor Suppressor Genes ◽

Methylation Status ◽

Prognostic Significance ◽

Promoter Hypermethylation ◽

Aberrant Methylation ◽

Suppressor Genes

Abstract Abstract 3950 Poster Board III-886 INTRODUCTION Germinal centers (GC) are unique sites in peripheral lymphoid tissue where clonal selection of B cells takes place. GC have been known to be a major source of B-cell lymphomas, including follicular (FL) and diffuse large cell (DLCL). DNA methylation of tumor-suppressor genes is a mechanism of gene silencing involved in the pathogenesis of FL and DLCBLs. Much less is known about the role of methylation in GCH. We determined the methylation status of 5 tumor-suppressor genes in 50 patients with lymphoma of GC origin and 50 GCH in order to find any differences between the pathological and the physiological state as well as its prognostic significance. MATERIAL AND METHODS. Genomic DNA extracted from paraffin-embedded samples of 30 DLCL, 20 FL and 50 GCH were analyzed by methylation-specific polymerase chain-reaction to determine promoter hypermethylation of DAP-k, SHP1, Rarβ, p14 and MGMT. Methylation status of each gene was correlated with clinicopathological status. Overall survival (OS) rates were calculated by the Kaplan-Meier method and differences were compared with the log-rank test. RESULTS. Median age was 65 in patients with lymphoma and 19 for GCH. Sex distribution was similar in all entities (60% females). Both lymphoma groups were balanced with respect to the presence of B symptoms, bulky disease, bone marrow infiltration, advanced stage and high IPI/FLIPI. DAP-k promoter methylation was present in more patients with lymphoma (89 and 87%) than with BFH (37%) p<0.0001. RaRB was methylated with higher frequency in FL (60%) than in DLCL (23%) and FH (12%) p<0.0001. SHP1 was more frequently methylated in FL (67%) and GCH (58%) than in DLCL (20%) p=0.01. Promoter hypermethylation of SHP1 was significantly associated with longer OS (p=0.021). Methylation of RaRB, p14 and MGMT were associated with shortened OS but the differences were not statistically significant. Those patients with DAPK methylated live longer but not significantly. In multivariate analysis hypermethylation of none of the genes studied remained an independent prognostic factor. CONCLUSIONS. Inactivation of DAP-K, and Rarβ is present in GC lymphomas with significantly higher frequency than in BFH. Thus, it may have pathogenic significance. SHP1 is methylated more frequently if FL and BFH than in DLCL, therefore that gene may be associated with aggressive disease. Methylation of DAP-k, SHP1, Rarβ, p14 and MGMT has no significant impact on overall survival. Markers for aberrant methylation may represent a promising way to monitor the onset and progression of malignancies but more extensive and prospective trials are needed to precisely define its role. Disclosures: No relevant conflicts of interest to declare.

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DNA methylation profiling in MEN1-related pancreatic neuroendocrine tumors reveals a potential epigenetic target for treatment

Acta Endocrinologica ◽

10.1530/eje-18-0195 ◽

2018 ◽

Vol 179 (3) ◽

pp. 153-160 ◽

Cited By ~ 5

Author(s):

E B Conemans ◽

L Lodewijk ◽

C B Moelans ◽

G J A Offerhaus ◽

C R C Pieterman ◽

...

Keyword(s):

Dna Methylation ◽

Tumor Suppressor ◽

Promoter Methylation ◽

Tumor Suppressor Genes ◽

Gene Promoter ◽

Pancreatic Neuroendocrine Tumor ◽

Promoter Hypermethylation ◽

Suppressor Gene ◽

Suppressor Genes ◽

Frequent Event

ObjectiveEpigenetic changes contribute to pancreatic neuroendocrine tumor (PanNET) development. Hypermethylation of promoter DNA as a cause of tumor suppressor gene silencing is a well-established oncogenic mechanism that is potentially reversible and therefore an interesting therapeutic target. Multiple endocrine neoplasia type 1 (MEN1) is the most frequent cause of inherited PanNETs. The aim of this study was to determine promoter methylation profiles in MEN1-related PanNETs.Design and methodsMethylation-specific multiplex ligation-dependent probe amplification was used to assess promoter methylation of 56 tumor suppressor genes in MEN1-related (n = 61) and sporadic (n = 34) PanNETs. Differences in cumulative methylation index (CMI), individual methylation percentages and frequency of promoter hypermethylation between subgroups were analyzed.ResultsWe found promoter methylation of a large number of potential tumor suppressor genes. CMI (median CMI: 912 vs 876,P = 0.207) was the same in MEN1-related and sporadic PanNETs. We found higher methylation percentages ofCASP8in MEN1-related PanNETs (median: 59% vs 16.5%,P = 0.002). In MEN1-related non-functioning PanNETs, the CMI was higher in larger PanNETs (>2 cm) (median: 969.5 vs 838.5;P = 0.021) and in PanNETs with liver metastases (median: 1036 vs 869;P = 0.013). Hypermethylation ofMGMT2was more frequent in non-functioning PanNETs compared to insulinomas (median: 44.7% vs 8.3%;P = 0.022). Hypermethylation of the Von Hippel–Lindau gene promoter was observed in one MEN1-related PanNET and was associated with loss of protein expression.ConclusionPromoter hypermethylation is a frequent event in MEN1-related and sporadic PanNETs. Targeting DNA methylation could be of therapeutic value in MEN1 patients with advanced PanNETs.

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TUMOR SUPPRESSOR GENES-THE EPIGENETIC BIOMARKERS IN HEPATOCELLULAR CARCINOMA

10.31219/osf.io/63gm2 ◽

2020 ◽

Author(s):

Debasray Saha

Keyword(s):

Tumor Suppressor ◽

Liver Cancer ◽

Tumor Suppressor Genes ◽

Promoter Hypermethylation ◽

Suppressor Gene ◽

Common Disease ◽

Epigenetic Changes ◽

Suppressor Genes ◽

Epigenetic Biomarkers ◽

Sequence Study

Majority of people infected from the cancer worldwide. Many types of cancer have been reported inprevious years and liver cancer one of them. 840,000 new cases of liver cancer have been reported worldwide in2018. Liver cancer is the 5th common disease in men and 9th in women. Smoking, chronic infection, hepatitis(hepatitis B & hepatitis C virus), liver cirrhosis, and genetic/epigenetic changes are most common risk factor forliver cancer. Like this, when gene change their expression without any transformation in DNA sequence study ofthis mechanism known as epigenetics. DNA methylation, histone modification, and RNA associate silencing aretype mechanism of epigenetic changes. Liver cancer can be caused by tumor suppressor gene (TSGs), and CpGIsland can lead TSGs gene silencing through promoter hypermethylation. A lot of hypermethylated gene has beenreported as a biomarker for prognosis of liver cancer. Some of these tumor suppressor genes such as BCL6B, RIZ1,SYK, GSTP1, DKK3, FPB1, TIP30, and ZHX2 are described through this paper.

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Consistent DNA Hypermethylation Patterns in Laryngeal Papillomas

International Journal of Head and Neck Surgery ◽

10.5005/jp-journals-10001-1013 ◽

2010 ◽

Vol 1 (2) ◽

pp. 69-77 ◽

Cited By ~ 8

Author(s):

Josena K Stephen ◽

Kang Mei Chen ◽

Veena Shah ◽

Vanessa G Schweitzer ◽

Glendon Gardner ◽

...

Keyword(s):

Dna Methylation ◽

Tumor Suppressor ◽

Tumor Suppressor Genes ◽

Methylation Status ◽

Promoter Hypermethylation ◽

Aberrant Methylation ◽

Dna Hypermethylation ◽

Suppressor Genes ◽

Multiple Tumor ◽

Specific Pcr

Abstract Introduction This study examined the contribution of promoter hypermethylation to the pathogenesis of respiratory papillomatosis (RP), including recurrences (RRP) and progression to squamous cell carcinoma (SSC). Materials and methods A retrospective cohort of 25 laryngeal papilloma cases included 21 RRP, two of which progressed to SCC. Aberrant methylation status was determined using the multigene (22 tumor suppressor genes) methylation-specific multiplex ligationdependent probe amplification assay and confirmed using methylation specific PCR. Results Twenty genes had altered DNA methylation in 22 of 25 cases. Aberrant methylation of CDKN2B and TIMP3 was most frequent. Promoter hypermethylation of BRCA2, APC, CDKN2A and CDKN2B was detected in 2 RRP cases with subsequent progression to SCC. Of the 25 cases, 22 were positive for HPV-6, 2 for HPV-11 and 1 for HPV-16 and 33. Conclusion Consistent aberrant methylation of multiple tumor suppressor genes contributes to the pathogenesis of laryngeal papillomas. Persistent aberrant DNA methylation events in 2 RRP cases that progressed to cancer indicate an epigenetic monoclonal progression continuum to SCC.

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