• BMB Reports
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• 제56권10호
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• pp.569-574
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• 2023

Aberrant DNA methylation plays a pivotal role in the onset and progression of colorectal cancer (CRC), a disease with high incidence and mortality rates in Korea. Several CRC-associated diagnostic and prognostic methylation markers have been identified; however, due to a lack of comprehensive clinical and methylome data, these markers have not been validated in the Korean population. Therefore, in this study, we aimed to obtain the CRC methylation profile using 172 tumors and 128 adjacent normal colon tissues of Korean patients with CRC. Based on the comparative methylome analysis, we found that hypermethylated positions in the tumor were predominantly concentrated in CpG islands and promoter regions, whereas hypomethylated positions were largely found in the open-sea region, notably distant from the CpG islands. In addition, we stratified patients by applying the CpG island methylator phenotype (CIMP) to the tumor methylome data. This stratification validated previous clinicopathological implications, as tumors with high CIMP signatures were significantly correlated with the proximal colon, higher prevalence of microsatellite instability status, and MLH1 promoter methylation. In conclusion, our extensive methylome analysis and the accompanying dataset offers valuable insights into the utilization of CRC-associated methylation markers in Korean patients, potentially improving CRC diagnosis and prognosis. Furthermore, this study serves as a solid foundation for further investigations into personalized and ethnicity-specific CRC treatments.

• Journal of Gastric Cancer
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• 제6권4호
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• pp.227-236
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• 2006

목적: 유전자 메틸화는 유전자의 서열에 영향을 주지 않으면서 유전자의 발현을 억제하고 세포분열 후 그대로 보존되는 후성적 변화이다. 위암조직과 정상위조직에서 hMLH1, p16, p14, COX-2, MGMT, E-cadherin 유전자와 MINT (MINT1, 2, 12, 25, 31)의 메틸화 상태를 검사하여 위암의 발생 과정에서의 작용과 CIMP 및 Helicobacter pylori균 감염을 포함한 임상병리학적인자와의 연관성을 알아보고자 하였다. 대상 및 방법: 위암과 정상위 신선 동결 조직 각각 36예를 대상으로 MSP (methylation-specific PCR)방법을 이용하여 메틸화 상태를 분석하였고 CIMP의 분석은 MINT1, MINT2, MINT12, MINT25, MINT31의 5개 marker를 대상으로 시행하였다. Helicobacter pylori균 감염여부는 Warthin-Starry silver 염색을 통하여 분류하였다. 결과: 위암 관련 유전자인 p14, p16, MGMT, COX-2, E-cadherin, hMLH1의 메틸화는 각각 14예(38.9%), 13예(36.1%), 8예(22.2%), 10예(27.8%), 21예(58.3%), 6예(16.7%)였다. MINT1과 MINT25의 메틸화는 위암조직에서 정상위조직에서보다 통계학적으로 유의하게 높게 관찰되었다. CIMP 양성률은 위암조직에서 44.4%로 높게 나타났으며 CIMP-H 위암은 환자의 연령과 종양크기와 연관이 있었다. CIMP 양성 위암은 p16 유전자의 메틸화와 연관이 있었고 p16 유전자의 메틸화는 조직학적으로 저분화, 미만형, 궤양형성하는 위암에서 낮게 나타났다. MINT1의 메틸화는 Helicobacter pylori균과 연관성이 있었다. 결론: 위암에서 hMLH1, p16, p14, COX-2, MGMT, E-cadherin, MINT (MINT1, 2, 12, 25, 31)의 불활성화에 DNA 메틸화가 작용함을 알 수 있었고, Helicobacter pylori균에 의한 위암발생에 MINT1의 메틸화가 연관이 있음을 알 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권2호
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• pp.541-549
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• 2015

Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer (BC) with higher metastatic rate and both local and systemic recurrence compared to non-TNBC. The generation of reactive oxygen species (ROS) secondary to oxidative stress is associated with DNA damage, chromosomal degradation and alterations of both hypermethylation and hypomethylation of DNA. This study concerns differential methylation of promoter regions in specific groups of genes in TNBC and non-TNBC Saudi females in an effort to understand whether epigenetic events might be involved in breast carcinogenesis, and whether they might be used as markers for Saudi BCs. Methylation of glutathione S-transferase P1 (GSTP1), T-cadherin (CDH13), Paired box protein 5 (PAX5), death associated protein kinase (DAPK), twist-related protein (TWIST), DNA-binding protein inhibitor (ID4), High In Normal-1 (HIN-1), cyclin-dependent kinase inhibitor 2A (p16), cyclin D2 and retinoic acid receptor-${\beta}$ ($RAR{\beta}1$) genes was analyzed by methylation specific polymerase chain reaction (MSP) in 200 archival formalin-fixed paraffin embedded BC tissues divided into 3 groups; benign breast tissues (20), TNBC (80) and non-TNBC (100). The relationships between methylation status, and clinical and pathological characteristics of patients and tumors were assessed. Higher frequencies of GSTP1, ID4, TWIST, DAPK, PAX5 and HIN-1 hypermethylation were found in TNBC than in non-TNBC. Hypermethylation of GSTP1, CDH13, ID4, DAPK, HIN-1 and PAX5 increased with tumor grade increasing. Other statistically significant correlations were identified with studied genes. Data from this study suggest that increased hypermethylation of GSTP1, ID4, TWIST, DAPK, PAX5 and HIN-1 genes in TNBC than in non-TNBC can act as useful biomarker for BCs in the Saudi population. The higher frequency of specific hypermethylated genes paralleling tumor grade, size and lymph node involvement suggests contributions to breast cancer initiation and progression.

• Molecules and Cells
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• 제24권3호
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• pp.364-371
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• 2007

The tumor suppressor gene Ras association domain family 1A (RASSF1A) is highly methylated in a wide range of human sporadic tumors. The current study investigated the hypermethylation of RASSF1A, the expression of RASSF1A protein, and the correlation between these and the clinicopathological features of gallbladder (GB) cancer in Korean patients. Formalin-fixed, paraffin-embedded tumors and non-neoplastic GB tissues (22 carcinomas, 8 adenomas, 26 normal epithelia) were collected from patients who had undergone surgical resection. The methylation status of two regions of the RASSF1A CpG island was determined by methylation-specific PCR (MSP), and the expression of RASSF1A protein was examined by immunohistochemistry using tissue microarrays. The K-RAS mutation was analyzed by direct sequencing. Methylation of the RASSF1A promoter (region 1) was detected in 22.7% (5/22) of carcinomas, 12.5% (1/8) of adenomas, and 0% (0/26) of normal gallbladder epithelia (P = 0.025). Methylation of the first exon (region 2) was found in 36.4% (8/22) of carcinomas, 25.0% (2/8) of adenomas, and 8.0% (2/26) of normal gallbladder epithelia (P = 0.038). K-RAS mutations were present in 4.5% (1/22) of carcinomas and 25% (2/8) of adenomas. RASSF1A methylaton was not associated with clinicopathological factors or K-ras mutation. Reduction or loss of RASSF1A expression was observed in most methylated adenocarcinomas. Three RASSF1A-expressing human biliary tract cancer cell lines examined contained unmethylated promoters and exons 1. These results suggest that downregulation of RASSF1A expression by DNA hypermethylation may be involved in GB carcinogenesis.

• 미생물학회지
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• 제44권4호
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• pp.277-281
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• 2008

HIV에 감염된 환자의 경우 다양한 종류의 암이 발생하는 것으로 알려져 있다. 이러한 암종의 높은 발생률의 원인으로, 감염에 의한 면역세포의 감소 및 결핍과 같은 간접적인 이유 뿐 아니라, HIV 바이러스 단백질의 발현이 직접적으로 병의 발생에 관여한다는 보고가 있다. 본 연구에서는 HIV 환자에서 높게 나타나는 암의 발생에 대한 기전을 이해하기 위하여 HIV-1 Tat 유전자와, 다수의 암 발생과 관련이 있는 세포막단백 CD99와의 관계를 규명하였다. 먼저 CD99의 발현에 미치는 Tat의 영향을 알아보기 위하여 HIV-1 Tat 발현 안정화 세포주를 확립하고 Tat 단백에 의한 CD99 유전자의 발현 양상 변화를 분석하였다. 실험결과 Tat의 발현에 의하여 CD99 유전자의 발현이 활성화되는 것이 관찰되었으며 이와 반대로 STAT3의 발현은 낮아졌다. CD99 프로모터는 CpG 함량이 높기 때문에 Tat 단백이 DNA 메칠화를 통해서 CD99 유전자의 발현을 조절하는지 확인하기 위하여 methylation specific PCR을 수행하였고 Tat의 발현이 높은 곳에서 특이적으로 CD99 프로모터 부위가 탈메칠화되는 것을 발견하였다. Tat 발현 세포에서만 특이적인 발현 차이를 보이는 유전자 분석을 위한 Differentially Expressed Gene keratin 17과 collagen, type IV 증가됨이 확인되었다. 위의 결과는 HIV Tat 단백이 직접 세포 단백들을 조절하여 암을 발생시킬 수 있다는 보고를 뒷받침한다.

• Tuberculosis and Respiratory Diseases
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• 제55권4호
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• pp.378-387
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• 2003

목 적 : 폐암의 발생 기전에서 종양 억제 유전자의 메틸화가 중요한 역할을 하는 것으로 알려져 있다. 본 연구는 폐암의 전이에 관여한다고 알려진 DAP kinase 유전자의 메틸화 양상을 폐암 환자의 말초 혈액 내 종양 DNA를 이용하여 알아보고자 하였다. 방 법 : 조직학적으로 윈발성 폐암으로 진단 받은 총 65명을 대상으로 말초 혈액 내에서 DNA를 추출하였다. 추출한 DNA를 Sodium bisulfite로 처리한 후 methylation specific PCR (MSP) 방법을 사용하여 DAP kinase 유전자의 비정상적인 메틸화 양상을 조사하였으며, 조직학적 양상, TNM병기, 임상 양상에 따른 비정상적인 메틸화 빈도의 차이성을 보았다. 결 과 : 전체 대상군은 남자 43명(65.2%), 여자 22명(33.8%)이었고 평균 연령은 6 1.0세, 평균 흡연력은 22.1갑/년이었다. 전체 65명 중 DAP kinase 유전자의 비정상적인 메틸화는 29명(44.6%)였다. 비소 세포암은 57명중 25명(43.9%), 소세포암은 8명중 4명(50.0%)에서 비정상적인 메틸화가 관찰되었으며, 조직형에 따른 통계학적 차이는 없었다. 비소세포암과 소세포암 모두에서 TNM 병기에 따른 비정상적인 메틸화 빈도의 차이는 없었다. 결 론 : 원발성 폐암 환자의 혈청에서 비교적 높은 빈도로 DAP kinase 유전자의 메틸화를 관찰할 수 있어 비침습적인 종양 표지자 검사로 이용될 수 있는 가능성을 보여 준다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권6호
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• pp.2753-2758
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• 2014

Background: GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. Materials and Methods: GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Results: Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4281-4287
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• 2014

Background: Methylation of tumor suppressor genes has been investigated in all kinds of cancer. Tumor specific epigenetic alterations can be used as a molecular markers of malignancy, which can lead to better diagnosis, prognosis and therapy. Therefore, the aim of this study was to evaluate the association between gene hypermethylation and expression of fragile histidine triad (FHIT), glutathione S-transferase P1 (GSTP1) and p16 genes and various clinicopathologic characteristics in primary non-small cell lung carcinomas (NSCLC). Materials and Methods: The study included 28 primary non-small cell lung carcinomas, where an additional 28 tissue samples taken from apparently normal safety margin surrounding the tumors served as controls. Methylation-specific polymerase chain reaction (MSP) was performed to analyze the methylation status of FHIT, GSTP1 and p16 while their mRNA expression levels were measured using a real-time PCR assay with SYBR Green I. Results: The methylation frequencies of the genes tested in NSCLC specimens were 53.6% for FHIT, 25% for GSTP1, and 0% for p16, and the risk of FHIT hypermethylation increased among patients with NSCLC by 2.88, while the risk of GSTP1 hypermethylation increased by 2.33. Hypermethylation of FHIT gene showed a highly significant correlation with pathologic stage (p<0.01) and a significant correlation with smoking habit and FHIT mRNA expression level (p<0.05). In contrast, no correlation was observed between the methylation of GSTP1 or p16 and smoking habit or any other parameter investigated (p>0.05). Conclusions: Results of the present study suggest that methylation of FHIT is a useful biomarker of biologically aggressive disease in patients with NSCLC. FHIT methylation may play a role in lung cancer later metastatic stages while GSTP1 methylation may rather play a role in the early pathogenesis.

• Molecules and Cells
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• 제45권12호
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• pp.886-895
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• 2022

Malignant rhabdoid tumor (MRT) is a highly aggressive pediatric malignancy with no effective therapy. Therefore, it is necessary to identify a target for the development of novel molecule-targeting therapeutic agents. In this study, we report the importance of the runt-related transcription factor 1 (RUNX1) and RUNX1-Baculoviral IAP (inhibitor of apoptosis) Repeat-Containing 5 (BIRC5/survivin) axis in the proliferation of MRT cells, as it can be used as an ideal target for anti-tumor strategies. The mechanism of this reaction can be explained by the interaction of RUNX1 with the RUNX1-binding DNA sequence located in the survivin promoter and its positive regulation. Specific knockdown of RUNX1 led to decreased expression of survivin, which subsequently suppressed the proliferation of MRT cells in vitro and in vivo. We also found that our novel RUNX inhibitor, Chb-M, which switches off RUNX1 using alkylating agent-conjugated pyrrole-imidazole polyamides designed to specifically bind to consensus RUNX-binding sequences (5'-TGTGGT-3'), inhibited survivin expression in vivo. Taken together, we identified a novel interaction between RUNX1 and survivin in MRT. Therefore the negative regulation of RUNX1 activity may be a novel strategy for MRT treatment.

• 생명과학회지
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• 제21권6호
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• pp.893-900
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• 2011

Prostatic acid phosphatase (PAP)는 전립선 암의 진단에 널리 사용되는 표지자로서 1935년 처음으로 동정되었고 인체 전립선에 가장 많이 존재하는 탈 인산화효소이다. PAP는 prostate epithelial cells에서 합성되는 전립선 특이적인 효소로서, 산성 환경에서 효소활성을 띠는 acid phosphatase 그룹에 속한다. PAP는 전립선액에 풍부히 존재하여 수정, 정자부족증, 만성통증의 감소에 관여한다. 그러나 가장 눈에 띄는 기능은 ERK1/2와 MAPK 경로에 관계된 HER-2와 PI3P의 탈 인산화를 유도하여 세포 성장 신호를 억제하고 전립선 암의 억제자로 작용하는 것이다. 최근 PAP DNA 백신을 이용하는 임상시험이 현재 진행 중이고, PAP를 이용한 immunotherapy를 통해 전립선 암을 치료하는 방법이 FDA의 승인을 받아 시행되고 있다. 이러한 PAP의 임상적 중요성에도 불구하고 현재까지 PAP의 분자적 조절기작에 대한 이해는 제한적이라 PAP에 대한 많은 연구가 필요한 실정이다. PAP는 NF-${\kappa}B$, TNF-${\alpha}$, IL-1 및 androgen과 androgen receptor에 의하여 promoter region이 조절된다고 알려졌다. 본 총설에서는 현재까지 밝혀진 PAP 유전자 및 단백질의 특징들과 더불어 전립선 암에서의 PAP의 기능, 발현 조절, 역할들을 종합하였다.