피부의 상피세포 종양에서 p53과 MDM2 발현에 대한 면역조직화학적 연구

조한석 ( Han Suk Cho ),박은주 ( Eun Joo Park ),김철우 ( Chul Woo Kim ),조희진 ( Hee Jin Cho ),김광호 ( Kwang Ho Kim ),김광중 ( Kwang Joong Kim ),박혜림 ( Hye Rim Park ) 대한피부과학회 2007 대한피부과학회지 Vol.45 No.6

Background: MDM2 (mouse double minute 2), the product of an oncogene is a key regulator of p53. It can repress transcription of p53 and eliminate it through proteolytic degradation. On the other hand, p53 binds specifically to the MDM2 and stimulates its transcription. Thus the two molecules are linked to each other through an autoregulatory feedback loop. Objective: To evaluate the expression patterns of p53 and MDM2 in several epithelial tumors and assess their correlations. Methods: We investigated the expression of p53 and MDM2 protein by an immunohistochemical method on formalin-fixed, paraffin-embedded tissue specimens of basal cell carcinoma (BCC), squamous cell carcinoma (SCC), Bowen`s disease, and actinic keratosis (AK). Results: The expression of p53 was increased in 83.3% (15/18) of BCC, 87.5% (14/16) of SCC, 75.0% (12/16) of Bowen`s disease, and 57% (8/14) of AK. The expression of MDM2 was increased in 50.0% (9/18) of BCC, 25.0%(4/16) of SCC, 68.8% (11/16) of Bowen`s disease, and 14.3% (2/14) of AK. The expression level of p53 had a significant positive correlation with the expression level of MDM2 in actinic keratosis (p=0.022). There was no correlation between the expression level of these two markers in others (p>0.05). Conclusion: This data suggests that p53 and MDM2 may be associated with pathogenesis of skin epithelial tumors with complex pathways. p53 (+)/MDM2 (+) phenotype expression may have an important pathogenetic role in Bowen`s disease. p53 (+)/MDM2 (-) type expression may have an important pathogenetic role in BCC, SCC, and AK. In particular, in AK p53 (+)/MDM2 (-) phenotype expression may have a major pathogenetic role. The mechanism of how they interact to promote tumorigenesis and the prognostic values in skin epithelial tumors remains to be elucidated though. (Korean J Dermatol 2007;45(6):534∼540)

두경부 편평상피세포암과 백반증에서 p53과 mdm-2의 면역조직화학적 연구

김용주,정환우,황찬승,양훈식 대한기관식도과학회 1998 大韓氣管食道科學會誌 Vol.4 No.1

The mutation of p53 is the most common genetic alteration found in human cancers and has oncogenic properties. mdm-2 is a recently discoverd that controls the p53 activity by binding of its protein, so negative feedback loop has been suggested in which p53 induces mdm-2 expression. The purpose of this study was to analyze the expression of p53 in leukoplakias, mdm-2 in squamous cell carcinomas, and relationship between p53 and mdm-2 expression in leukoplakias and squamous cell carcinomas. The results were as follows : 1) The p53 was expressed 33.4% in leukoplakias 2) The mdm-2 was expressed 8.3% in leukoplakias and 22.7% in squamous cell carcinomas. 3) The expression rate of p53 was higher in specimens negative for mdm-2 than in specimens positive for mdm-2, but there was not significant relationship between p53 and mdm-2 expression. In conclusion p53 was thought to participate in early phase of oncogenesis, and mdm-2 was thought to have a role as a oncogene in squamous cell carcinoma of head and neck. Though there was not significant relationship between p53 and mdm-2 expression, mdm-2 was thought to inhibit p53 activity.

식도암에서 MDM2, p53, pRb 발현과 동시적 항암화학방사선요법의 결과

윤미선(Mee Sun Yoon),남택근(Taek Keun Nam),이재혁(Jae Hyuk Lee),조상희(Sang Hee Cho),송주영(Ju Young Song),안성자(Sung Ja Ahn),정익주(Ik Joo Chung),정웅기(Woong Ki Chung),나병식(Byung Sik Nah) 대한방사선종양학회 2007 Radiation Oncology Journal Vol.25 No.4

목 적: 식도암에서 동시적 항암화학방사선치료 전 MDM2, p53, pRb 발현양상이 치료반응 및 생존율 등 치료결과와 연관성이 있는지 알아보고자 하였다. 대상 및 방법: AJCC 병기 I~IVa로 근치적 목적의 동시적 항암화학방사선요법을 받은 51명의 환자를 대상으로 하였다. 방사선치료는 일일 1.8~2.0 Gy씩 원발병소에 중앙값 54 Gy를 시행하였고 항암화학요법은 CDDP/5-FU를 4주 간격으로 4회 시행하고 첫 2회는 방사선치료와 동시에 시행하였다. MDM2, p53, pRb 발현의 검출은 치료 전 내시경하 조직생검을 이용하여 면역조직화학 염색방법을 이용하였다. 단백발현 양성종양세포가 50% 이상인 경우를 고발현군으로 정의하였다. 결 과: 전체 환자의 중앙 추적관찰기간은 26개월이었다. MDM2, p53, pRb 고발현군은 각각 19.6%, 27.5%, 66.7% 이었다. 그러나 이들의 발현 정도와 치료반응, 종양특이 생존율, 전체 생존율 등 모두 유의한 연관성은 없었다. 연령(65세 이하 vs. 초과), 종양의 위치(상부, 중앙부, 하부), 종양의 길이(5 cm 이하 vs. 초과), 병기(I∼II vs. III~IVa), MDM2 (저발현 vs. 고발현), p53 (저발현 vs. 고발현), pRb (저발현 vs. 고발현), 병리학적 완전관해여부, 임상적 완전관해여부 등 9개 요인들을 대상으로 종양특성 생존율에 대한 다변량 분석에서 병리학적 완전관해여부(RR12.100, p<0.001)와 병기(RR 3.300, p=0.028) 만이 유의한 인자들이었다. 결 론: 본 연구에서 MDM2, p53, pRb의 치료 전 발현양상과 치료결과와 의미있는 연관성은 발견할 수 없었다. 향후상기 발현인자들을 포함하여 잠재적인 예후인자로서 새로운 다른 발현인자들을 발굴하고 보다 많은 증례 수를 대상으로 추적기간을 보강하여 이들을 재평가하는 연구가 요망된다. Purpose: This study evaluated the pretreatment expression patterns of MDM2, p53, and pRb proteins to determine if the expression patterns could predict the outcome of concurrent chemoradiotherapy (CCRT) for esophageal squamous cell carcinoma and aid in the decisions for the selection of treatment modalities. Materials and Methods: Fifty-one patients that were treated with definitive chemoradiotherapy for stage I∼IVa esophageal squamous cell carcinoma were selected for this study. Radiotherapy was administered with daily 1.8 ∼2 Gy fractions up to a median dose of 54 Gy for primary tumors, and with four cycles of cisplatin/5- fluorouracil chemotherapy that was administered every 4 weeks, the first two cycles of which were administered concurrently with radiotherapy. Expression of MDM2, p53, and pRb was investigated by immunohistochemical analysis using pretreatment biopsy specimens. Results: MDM2, p53, and pRb were detected with high immunoreactivity in 19.6%, 27.5%, and 66.7% of the patients, respectively. However, there was no significant correlation between expression of these factors and clinical outcome. By the use of multivariate analysis with nine covariates-age, tumor location, tumor length, stage, pathological response, clinical response, MDM2 expression, p53 expression, and pRb expression, only pathological response and stage were significant factors for cause-specific survival. Conclusion: Expression of MDM2, p53, and pRb was not found to be clinically significant for predicting outcomes after CCRT in this study. Further studies with a larger patient population and longer follow-up periods are needed to re-evaluate the expression pattern and to identify new predictors for CCRT response.

Role of the MDM2 Promoter Polymorphism (-309T>G) in Acute Myeloid Leukemia Development

Cingeetham, Anuradha,Vuree, Sugunakar,Jiwatani, Sangeeta,Kagita, Sailaja,Dunna, Nageswara Rao,Meka, Phanni Bhushann,Gorre, Manjula,Annamaneni, Sandhya,Digumarti, Raghunadharao,Sinha, Sudha,Satti, Vish Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.7

Background: The human homologue of the mouse double minute 2 (MDM2) gene is a negative regulator of Tp53. MDM2-309T>G a functional promoter polymorphism was found to be associated with overexpression thereby attenuation of Tp53 stress response and increased cancer susceptibility. We have planned to evaluate the possible role of MDM2-309T>G polymorphism with risk and response to chemotherapy in AML. Materials and Methods: A total of 223 de novo AML cases and 304 age and sex matched healthy controls were genotyped for the MDM2-309T>G polymorphism through the tetra-primer amplification refractory mutation system (ARMS)-PCR method. In order to assess the functional relationship of -309T>G SNP with MDM2 expression level, we quantified MDM2 mRNA in 30 primary AML blood samples through quantitative RT-PCR. Both the (-309T>G) genotypes and the MDM2 expression were correlated with disease free survival (DFS) rates among patients who have achieved complete remission (CR) after first induction chemotherapy. Results: MDM2-309T>G polymorphism was significantly associated with AML development (p<0.0001). The presence of either GG genotype or G allele at MDM2-309 confered 1.79 (95% CI: 1.12-2.86; p<0.001) and 1.46 fold (95%CI: 1.14-1.86; p= 0.003) increased AML risk. Survival analysis revealed that CR+ve cases with GG genotype had significantly increased DFS rates (16months, p=0.05) compared to CR+ve TT (11 months) and TG (9 months) genotype groups. Further, MDM2 expression was also found to be significantly elevated in GG genotype patients (p=0.0039) and among CR+ve cases (p=0.0036). Conclusions: The MDM2-309T>G polymorphism might be involved in AML development and also serve as a good prognostic indicator.

다발성골수종 환자에서 p53, MDM2, Cyclin D1 단백 발현의 임상적 유용성

장은아,현봉학,이기범,차영주 大韓血液學會 2000 大韓血液學會誌 Vol.35 No.3

피부 악성 종양에서의 mdm - 2 단백 표현에 관한 면역조직화학적 연구

조지형 ( Ji Hyeung Cho ),노영석 ( Young Suck Ro ) 대한피부과학회 1996 대한피부과학회지 Vol.34 No.4

Background: The human homologue of mouse double minute 2(mdm-2) gene codes for a cellular protein that forms a complex with the mutant and wild type p53 protein and modulates its trans-activation activity. Overexpression of the mdm-2 protein is associated with the tumorigenic potential of mdm-2 gene and overcomes the growth suppressive activity of p53. Objective & Methods : In order to investigate the pattern of the mdm-2 and p53 expression in malignant skin tumors, we performed an immunohistochemical analysis using NCL-MDM-2p polyclonal antibody and DO7 monoclonal antibody, respectively, in 10 squamous cell carcinomas (SCCs), 8 basal cell carcinomas(BCCs), 5 malignant melanomas(MMs), and 6 fibrosarcomas (FSs). Results : The expression of mdm-2 protein was detected in 40% of SCCs, 25% of BCCs, 40% of MMs and 33% of FSs, whereas 40% of SCCs, 25% of BCCs, 20% of MMs, and 17% of FSs showed p53 positivity. One case of SCCs and one case of BCCs showed immunoreactivity with both anti-mdm-2 and anti-p53 antibodies. Within the limitations of this small sample size, there was no relationship between the expression of mdm-2 and p53 protein, as overexpression of these two genes was not mutually exclusive. Conclusion : These findings suggest that interaction between the mdm-2 and p53 genes and products in these malignant tumors appears to be heterogenous and more complex than previously realized. Further studies on molecular basis are required to characterize the mechanism of mdm-2 overexpression and its relation to p53. (Kor J Dermatol 1996;34(4): 538-545)

자궁경부 편평상피암종에서 p53 단백과 MDM-2 단백 발현에 관한 면역 조직화학 연구

정찬우(Chan Woo Chung),신용각(Yong Kag Shin),박채웅(Chae Woong Park),김창주(Chang Joo Kim),조태일(Tae Il Cho),박언섭(Eon Sub Park) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.10

목적: 자궁경부 상피내병변 및 침윤성 암종에서 p53 단백과, p53과 길항적으로 작용한다고 알려진 MDM-2 단백발현과 더불어 상호간의 관련성을 면역조직화학적으로 관찰하였다. 연구방법: 중앙대학교 부속병원에서 자궁경부 편평상피암종 또는 상피내 종양으로 자궁경부 원추절제술, 단순 또는 근치적 절제수술을 받은 조직중 표본 상태가 양호한 62예를 선택하였다. 병리조직학적 분류는 WHO의 분류에 따라 상피내 종양과 침윤성 암종으로 분류하였고, 상피내 종양은 다시 CINⅠ,Ⅱ,그리고 CINⅢ로 분류하였고, 각각의 증례에 p53, MDM-2 항체에 대한 면역조직화학 염색방법을 실시 하였다. 결과: p53 발현은 경도 또는 중등도 이형증식(CINⅠ과 CINⅡ)에서 25%, 고도 또는 상피내 암종에서 20%, 침윤성 암종에서는 44%의 발현률을 보여 주었다. MDM-2 발현은 경도 또는 중등도 이형증식에서 33%, 고도 또는 상피내 암종에서 16%, 침윤성 암종에서는 48%에서 관찰되었다. p53 양성 발현과 MDM-2 양성 발현간에는 의미있는 상관관계가 없었다(p>0.05). 그렇지만 MDM-2 음성인 군에서 통계적으로 유의하게 p53 단백발현이 음성으로 나타났다(p=0.002). 결론: 자궁경부의 전암성 및 침윤성 암종에서 발현되는 p53 기능장애의 한 원인 인자로 MDM-2 과발현이 관여하지만 MDM-2 발현 이외에 다른 인자도 관여 할 것이라 생각되었다. Objectives: MDM-2 is an oncoprotein that inhibits p53 tumor suppressor protein. Amplication and over- expression of its protein have been observed in human malignancies, and these abnormalities have a role in tumorigenesis through inactivation of p53 function. To elucidate the role of p53 and MDM-2 protein in cervical neoplasia we investigated the expression rates of MDM-2 and p53 protein in surgically resected specimens. Metheds: Immunohistochemical studies using anti-p53 and anti-MDM-2 protein in the paraffin embedded section of 62 cases including cervical intraepithelial neoplasm(CIN) and invasive cervical cancer were performed. Results: Expression rates of p53 protein were 25% in CINⅠ& CINⅡ, 20% in CINⅢ, and 44% in invasive carcinoma, respectively. The MDM-2 protein were 33% in CINⅠ& CINⅡ, 16% in CINⅢ, and 48% in invasive carcinoma, respectively. There was no evident correlation between p53 positivity and MDM-2 positivity(p>0.05). However, correlation between MDM-2 negativity and p53 negativity was statistically significant(p=0.002) Conclusion: These data suggest that the expression of p53 protein is presumed to be necessarily correlated with MDM-2 protein expression in cervical neoplasia.

MDM2 Expression in Serous and Mucinous Epithelial Tumours of the Ovary

Abdelaal, Shereen E,Habib, Fahima M,el Din, Amina A Gamal,Gabal, Samia M,Hassan, Nabila S,Ibrahim, Nihad A Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.7

Background: Different types of cancer exhibit abnormalities in cell cycle regulators. The murine double minute-2(MDM2) cell cycle regulator is a proto-oncogene that negatively regulates the P53 tumour suppressor gene. Surface epithelial tumours constitute approximately two thirds of ovarian neoplasms. Each histologic type can be classified as benign, borderline and malignant. This study aimed to examine immunohistochemical expression of the MDM2 protein in ovarian serous and mucinous epithelial tumours (benign, borderline and malignant). Materials and Methods: This study included forty five ovarian tumours, subdivided into fifteen cystadenomas (5 serous and 10 mucinous), fifteen borderline tumours (11 serous and 4 mucinous) and fifteen cystadenocarcinomas (9 serous and 6 mucinous). Paraffin sections were stained with haematoxylin and eosin for histopathologic study, and with mouse monoclonal anti-MDM2 antibody for immunohistochemistry. Results: MDM2 positivity was detected in 28.9% of the studied ovarian tumours. All benign tumours were negative and positivity was significantly higher in malignant than borderline tumours (P value of chi-square test =0.000). Significantly, all MDM2 positive mucinous tumours were malignant with no positive mucinous borderline tumours. Malignant tumours showed positive MDM2 expression in 83.3% of mucinous type and in 55.6% of serous type. Borderline serous tumours showed negative MDM2 in 72.7% of cases (P value of Z test =0.04). Conclusions: Alterations in the expression of the cell cycle regulator (MDM2) occur early in the process of tumourigenesis in serous and mucinous ovarian tumours. We suggest that MDM2 may be used in those tumours as a marker for risk stratification and identification of cases with cancer development and progression. We recommend further studies on MDM2 immunohistochemistry, in conjunction with adjuvant methods as DNA ploidy and FISH gene amplification, focusing on the mucinous tumours and differentiating between the three tumour categories, benign, borderline and malignant.

Correlation between Patterns of Mdm2 SNIP 309 and Histopathological Severity of Helicobacter pylori Associated Gastritis in Thailand

Tongtawee, Taweesak,Dechsukhum, Chavaboon,Talabnin, Krajang,Leeanansaksiri, Wilairat,Kaewpitoon, Soraya,Kaewpitoon, Natthawut,Loyd, Ryan A,Matrakool, Likit,Panpimanmas, Sukij Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.17

Background: The commonly held view of the tumor suppressor p53 is as a regulator of cell proliferation, apoptosis and many other biological processes as well as external and internal stress responses. Mdm2 SNIP309 is a negative regulator of p 53. Therefore, this study aimed to determine the correlation between the patterns of Mdm2 SNIP 309 and the inflammation grading of Helicobacter pylori associated gastritis in a Thai population. Materials and Methods: A cross-sectional study was carried out from November 2014 through June 2015. Biopsy specimens were obtained from infected patients and infection was proved by positive histology. The gastric mucosa specimens were sent to the Molecular Genetic Unit, Institute of Medicine, Suranaree University of Technology where they were tested by molecular methods to detect the patterns of Mdm2 SNIP 309 using the real-time PCR hybridization probe method. The results were analyzed and compared with the Updated Sydney classification. Results: A total of 100 infected patients were interviewed and gastric mucosa specimens were collected. In this study the percentage of Mdm2 SNIP 309 T/T homozygous and Mdm2 SNIP309 G/T heterozygous was 78% and 19 % respectively whereas Mdm2 SNIP309 G/G homozygous was 3%. Mdm2 SNIP 309 T/T homozygous and Mdm2 SNIP309 G/T heterozygous correlated with mild to moderate inflammation (P<0.01) whereas Mdm2 SNIP309 G/G homozygous correlated with severe inflammation (P<0.01). Conclusions: Our study found the frequency of Mdm2 SNP309 G/G in our Thai population to be very low, and suggests that this can explain to some extent the low incidence of severe inflammation and gastric cancer changes in the Thai population. Mild to moderate inflammation are the most common pathologic gradings due to the unique genetic polymorphism of Mdm2 SNIP 309 in the Thai population.

Role of the Mdm2 SNIP 309 Polymorphism in Gastric Mucosal Morphologic Patterns of Patients with Helicobacter pylori Associated Gastritis

Tongtawee, Taweesak,Dechsukhum, Chavaboon,Leeanansaksiri, Wilairat,Kaewpitoon, Soraya,Kaewpitoon, Natthawut,Loyd, Ryan A,Matrakool, Likit,Panpimanmas, Sukij Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.3

Background: The tumor suppressor p53 is as a regulator of cell proliferation, apoptosis and many other biological processes as well as external and internal stress responses. Mdm2 SNIP309 is a negative regulator of 53. Therefore, this study aimed to determine the role of the Mdm2 SNIP 309 polymorphism in the gastric mucosal morphological patterns in patients with Helicobacter pylori associated gastritis. Materials and Methods: A prospective cross-sectional study was carried out from November 2014 through November 2015. Biopsy specimens were obtained from patients and infection was proven by positive histology. Gastric mucosa specimens were sent to the Molecular Genetics Unit, Institute of Medicine, Suranaree University of Technology where they were tested by molecular methods to detect the patterns of Mdm2 SNIP 309 polymorphism using the real-time PCR hybridization probe method. The results were analyzed and correlated with gastric mucosal morphological patterns by using C-NBI endoscopy. Results: A total of 300 infected patients were enrolled and gastric mucosa specimens were collected. In this study the percentage of Mdm2 SNIP 309 T/T homozygous and Mdm2 SNIP309 G/T heterozygous was 78% and 19 % respectively whereas Mdm2 SNIP309 G/G homozygous was 3%. Mdm2 SNIP 309 T/T homozygous and Mdm2 SNIP309 G/T heterozygosity correlated with type 1 to type 3 gastric mucosal morphological patterns (P<0.01) whereas Mdm2 SNIP309 G/G homozygous correlated with type 4 and type 5 (P<0.01). Conclusions: Our study finds the frequency of Mdm2 SNIP309 G/G in a Thai population is very low, and suggests that this can explain ae Thailand enigma. Types 1 to type 3 are the most common gastric mucosal morphological patterns according to the unique genetic polymorphism of MDM2 SNIP 309 in the Thai population.