비가족성 산발성 결직장암에서 Microsatellite Instability 와 병리조직학적 특징 및 p53 단백질 발현과의 연관성

양영일,김미성,윤혜경,임인숙 인제대학교 1997 仁濟醫學 Vol.18 No.4

Microsatellite instability는 선천성 비용종성 결직장암의 분자생물학적 원인 인자로 규명되었으며 산발성 결직장암에서도 원인 인자로 그 중요성이 제시되었다. 가족력이 동반되지 않은 산발성 결직장암 49예를 대상으로 Microsatellite instability의 빈도를 조사하여 병리학적 특성, 세포 증식력, p53 단백질 발현과의 연관성을 연구하였다. 비가족성 산발성 결직장암에서 Microsatellite instability는 24.5%에서 검출되었고, Microsatellite instability 양성 결직장암은 점액성 분화, 종양 주위의 기질내에 림프구 집합의 형성이 빈번히 동반되는 병리조직학적 특성들을 보였다. p53 단백질 발현은 microsatellite instability 양성 결직장암에서 양성율이 16.7%인 반면 음성인 종양인 경우는 67.6%로 p53 단백질 발현과 microsatellite instability 사이에는 의의 있는 역상관 관계를 보였다. 이상의 결과로 Microsatellite instability는 결직장암의 발생 과정 및 형태학적 형성 과정에 중요한 역할을 담당할 것으로 사료된다. Microsatellites are highly polymorphic, short-tandem repeat sequences dispersed throughout the genome. Defects in DNA mismatch repair can result in an increase in DNA replication errors (RER) or microsatellite instability (MSI), which are manifested as extra, aberrant bands within multiple microsatellite loci. MSI are found in the cancers from patients with hereditary non-polyposis colorectal cancer and have been also demonstrated in sporadic colorectal carcinomas (CRC). It has been suggested that the histologic features and molecular characteristics of MSI-positive CRC are similar to those of HNPCC. In order to clarify the relationship between MSI and pathological parameters including histologic features, proliferating activity, p53 protein expression, PCNA proliferating activity and PCR for microsatellite instability was performed on a series of 49 formalin-fixed CRC and normal mucosa. (1) MSI-positive CRC's were noted in 12 (24.5%) CRC cases, but no MSI was detected in normal mucosal tissues. (2) MSI-positive CRC's were closed related with extracellular mucin production, plexiform pattern, and Crohns-like lymphoid reaction (p<0.05). (3) No significant relationship between MSI and age, sex, tumor size, location, grade, stage, vascular invasion and PCNA index was noted (p<0.05). (4) p53 protein expression rate of 49 CRC cases was 55.1%, and p53 expression rate of MSI-negative group was 67.6% and the rate in MSI-positive group was 16.7%. The inverse relationship between MSI and p53 expression was statistically significant (p<0.05). In summary, microsatellite instability may play an important role in colorectal carcinogenesis and morphogenesis of non-familial colorectal carcinomas, and inverse correlation between MSI phenotype and p53 expression suggests MSI and p53 mutations are independent pathway in colorectal carcinogenesis. And MSI-associated unique pathologic features may be useful for screening criteria of patients wish HNPCC.

Carcinoma Microsatellite Instability Status as a Predictor of Benefit from Fluorouracil-Based Adjuvant Chemotherapy for Stage II Rectal Cancer

Yang, Liu,Sun, Yan,Huang, Xin-En,Yu, Dong-Sheng,Zhou, Jian-Nong,Zhou, Xin,Li, Dong-Zheng,Guan, Xin Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.4

Purpose: Rectal cancers with high microsatellite-instable have clinical and pathological features that differentiate them from microsatellite-stable or low-frequency carcinomas, which was studied rarely in stage II rectal cancer, promoting the present investigation of the usefulness of microsatellite-instability status as a predictor of the benefit of adjuvant chemotherapy with fluorouracil in stage II rectal cancer. Patients and Methods: Data of 460 patients who underwent primary anterior resection with a double stapling technique for rectal carcinoma at a single institution from 2008 to 2012 were retrospectively collected. All patients experienced a total mesorectal excision (TME) operation. Survival analysis were analyzed using the Cox regression method. Results: Five-year rate of disease-free survival (DFS) was noted in 390 (84.8%) of 460 patients with stage II rectal cancer. Of 460 tissue specimens, 97 (21.1%) exhibited high-frequency microsatellite instability. Median age of the patients was 65 (50-71) and 185 (40.2%) were male. After univariate and multivariate analysis, microsatellite instability (p= 0.001), female sex (p<0.05) and fluorouracil-based adjuvant chemotherapy (p<0.001), the 3 factors were attributed to a favorable survival status independently. Among 201 patients who did not receive adjuvant chemotherapy, those cancers displaying high-frequency microsatellite instability had a better 5-year rate of DFS than tumors exhibiting microsatellite stability or low-frequency instability (HR, 13.61 [95% CI, 1.88 to 99.28]; p= 0.010), while in 259 patients who received adjuvant chemotherapy, there was no DFS difference between the two groups (p= 0.145). Furthermore, patients exhibiting microsatellite stability or low-frequency instability who received adjuvant chemotherapy had a better 5-year rate of DFS than patients did not (HR, 5.16 [95% CI, 2.90 to 9.18]; p<0.001), while patients exhibiting high-frequency microsatellite instability were not connected with increased DFS (p= 0.696). It was implied that female patients had better survival than male. Conclusion: Survival status after anterior resection of rectal carcinoma is related to the microsatellite instability status, adjuvant chemotherapy and gender. Fluorouracil-based adjuvant chemotherapy benefits patients of stage II rectal cancer with microsatellite-stable or low microsatellite-instable, but not those with high microsatellite-instable. Additionally, free of adjuvant chemotherapy, carcinomas with high microsatellite-instable have a better 5-year rate of DFS than those with microsatellite-stable or low microsatellite-instable, and female patients have a better survival as well.

Frameshift mutations of chromosome cohesion-related genes SGOL1 and PDS5B in gastric and colorectal cancers with high microsatellite instability

Kim, M.S.,An, C.H.,Yoo, N.J.,Lee, S.H. W. B. Saunders Co ; Centrum Philadelphia 2013 Human pathology Vol.44 No.10

Cohesin is a protein complex that regulates chromatid cohesion and plays a role in preventing aneuploidy and maintaining chromosomal stability. SGOL1 encodes a cohesin protector, and PDS5B encodes a regulatory cohesion factor. Both SGOL1 and PDS5B are considered putative tumor suppressor genes. The aim of this study was to explore whether SGOL1 and PDS5B genes are mutated and expressionally altered in gastric and colorectal cancers. A genome database indicated that both genes possessed mononucleotide repeats in coding sequences, which could be mutation targets in cancers with microsatellite instability. We analyzed mutations in 91 gastric cancers and 100 colorectal cancers with high microsatellite instability or stable/low microsatellite instability by single-strand conformation polymorphism analysis and DNA sequencing. We also analyzed SGOL1 and PDS5B expression by immunohistochemistry. Overall, we found 21 SGOL1 frameshift mutations in 21 cases and 18 PDS5B frameshift mutations in 16 cases. SGOL1 and PDS5B frameshift mutations were detected in 26.6% and 20.3%, respectively, of high microsatellite instability but not in stable/low microsatellite instability (0/112). By immunohistochemistry, losses of SGOL1 and PDS5B were identified in 19% to 47% of the gastric and colorectal cancers irrespective of microsatellite instability status. The losses were more common in those with frameshift mutations or high microsatellite instability than those without mutations or high microsatellite instability. The data indicate that frameshift mutations of SGOL1 and PDS5B and the loss of their expression may be a feature of gastric and colorectal cancers with high microsatellite instability. In addition, the data suggest that these alterations might contribute to cancer pathogenesis by deregulating cohesin-related functions.

산발성 장형 위선암 환자의 Microsatellite Instability와 병리학적 양상

조창희,홍유찬,안지현,최경현,이상호,신영명,윤기영,정민정,장희경 고신대학교의과대학 2007 고신대학교 의과대학 학술지 Vol.22 No.2

Background : Through many researches, microsatellite is expected to be a good diagnositic and prognostic factor in colorectal cancer, endometrial cancer, gastric cancer, and the others. The prevalence of microsatellite instability (MSI) in gastric carcinoma has reported variously, 13~44%. Purpose : We aimed to determine the prevalence of MSI-high and the relationship between MSI and pathological characteristics of sporadic intestinal type adenocarcinoma of stomach. Material and Methods : We analyzed 106 sporadic intestinal type adenocarcinoma specimens excised from patients who were over thirty-five years old to determine the statue of microsatellite by DNA sequencing. The tissues were formalin-fixed and paraffin embedded. DNA were extracted and amplified by polymerase chain reaction (PCR). MSI was determined using five markers recommended by National Cancer Institute (NCI). Specimens were also studied with five patholical factors-differenciation of tumor cells, depth of invasion, lymph node metastasis, vessel invasion, and perineural invasion- to determine pathological state. Result : The microsatellite statue was determined as MSI-High in 5 cases (4.7%), no MSI-low, and MSS (microsatellite stable) in 101 cases (95.3%). Within the frequency, there was no large gap in the distinction of gender in MSI cases, but in MSS cases, there was three-times more cases in male. MSI cases had moderate-to-poor differenciation and trend to invade toward serosa. All MSI cases showed no perineural invasion. But we could not find any statistical significance between MSI and pathological characteristics of sporadic intestinal type adenocarcinoma. Conclusion : Results suggest that MSI can not make any certain pathological significance in sporadic intestinal type adenocarcinoma. Even though less than 5% of sporadic intestinal type adenocarcinoma patients showed MSI, it can be used as a influential prediagnostic factor of gastric cancer. Further study with large scale of cases will be followed to verify these results.

Frameshift mutations of tumor suppressor gene EP300 in gastric and colorectal cancers with high microsatellite instability

Kim, M.S.,Lee, S.H.,Yoo, N.J.,Lee, S.H. W. B. Saunders Co ; Centrum Philadelphia 2013 Human pathology Vol.44 No.10

Several lines of evidence show that chromatin remodeling is involved in the pathogenesis of disease, including cancer. The E1A-binding protein p300 functions as a histone acetyltransferase and is considered an important modulator of chromatin remodeling. The aim of this study was to explore whether E1A-binding protein p300 is somatically mutated and expressionally altered in gastric and colorectal cancers. By analyzing a public database, we found that E1A-binding protein p300 had mononucleotide repeats in exons 27 and 31 that could be mutation targets in cancers with microsatellite instability. We analyzed mutations in the mononucleotide repeats in 91 gastric and 101 colorectal cancers with high microsatellite instability or stable microsatellite instability by single-strand conformation polymorphism analysis and DNA sequencing. We also analyzed E1A-binding protein p300 expression in gastric and colorectal cancers by immunohistochemistry staining. We found E1A-binding protein p300 frameshift mutations (4 in exon 27 and 3 in exon 31) in 3 gastric and 4 colorectal cancers that were detected exclusively in cancers with high microsatellite instability (7/80). In the immunohistochemistry study, loss of E1A-binding protein p300 expression was identified in 12% and 24% of the gastric and colorectal cancers, respectively, irrespective of microsatellite instability status. Loss was more common in tumors with E1A-binding protein p300 frameshift mutations. Frameshift mutations of E1A-binding protein p300 and its expressional loss may be a feature of gastric and colorectal cancers with high microsatellite instability. These alterations could contribute to cancer pathogenesis by deregulating E1A-binding protein p300-mediated functions.

Clinical Implications of Microsatellite Instability in Early Gastric Cancer

Kim, Dong Gyu,An, Ji Yeong,Kim, Hyunki,Shin, Su-Jin,Choi, Seohee,Seo, Won Jun,Roh, Chul Kyu,Cho, Minah,Son, Taeil,Kim, Hyoung-Il,Cheong, Jae-Ho,Hyung, Woo Jin,Noh, Sung Hoon,Choi, Yoon Young The Korean Gastric Cancer Association 2019 Journal of gastric cancer Vol.19 No.4

Purpose: We aimed to evaluate the clinical characteristics of microsatellite instability in early gastric cancer. Materials and Methods: The microsatellite instability status of resected early gastric tumors was evaluated using two mononucleotide repeat markers (BAT25 and BAT26) and three dinucleotide repeat markers (D5S346, D2S123, and D17S250). Tumors with instability in two or more markers were defined as microsatellite instability-high (MSI-H) and others were classified as microsatellite stable (MSS). Results: Overall, 1,156 tumors were included in the analysis, with 85 (7.4%) classified as MSI-H compared with MSS tumors. For MSI-H tumors, there was a significant correlation with the female sex, older age, tumor location in the lower gastric body, intestinal histology, lymphovascular invasion (LVI), and submucosal invasion (P<0.05). There was also a trend toward an association with lymph node (LN) metastasis (P=0.056). In mucosal gastric cancer, there was no significant difference in MSI status in tumors with LN metastasis or tumors with LVI. In submucosal gastric cancer, LVI was more frequently observed in MSI-H than in MSS tumors (38.9% vs. 25.0%, P=0.027), but there was no difference in the presence of LN metastases. The prognosis of MSI-H tumors was similar to that of MSS tumors (log-rank test, P=0.797, the hazard ratio for MSI-H was adjusted by age, sex, pT stage, and the number of metastatic LNs, 0.932; 95% confidence interval, 0.423-2.054; P=0.861). Conclusions: MSI status was not useful in predicting prognosis in early gastric cancer. However, the frequent presence of LVI in early MSI-H gastric cancer may help guide the appropriate treatment for patients, such as endoscopic treatment or limited LN surgical dissection.

Clinical Implications of Microsatellite Instability in Early Gastric Cancer

최윤영,Dong Gyu Kim,안지영,김현기,Su-Jin Shin,Seohee Choi,Won Jun Seo,Chul Kyu Roh,Minah Cho,손태일,김형일,정재호,형우진,노성훈 대한위암학회 2019 Journal of gastric cancer Vol.19 No.4

Purpose: We aimed to evaluate the clinical characteristics of microsatellite instability in early gastric cancer. Materials and Methods: The microsatellite instability status of resected early gastric tumors was evaluated using two mononucleotide repeat markers (BAT25 and BAT26) and three dinucleotide repeat markers (D5S346, D2S123, and D17S250). Tumors with instability in two or more markers were defined as microsatellite instability-high (MSI-H) and others were classified as microsatellite stable (MSS). Results: Overall, 1,156 tumors were included in the analysis, with 85 (7.4%) classified as MSI-H compared with MSS tumors. For MSI-H tumors, there was a significant correlation with the female sex, older age, tumor location in the lower gastric body, intestinal histology, lymphovascular invasion (LVI), and submucosal invasion (P<0.05). There was also a trend toward an association with lymph node (LN) metastasis (P=0.056). In mucosal gastric cancer, there was no significant difference in MSI status in tumors with LN metastasis or tumors with LVI. In submucosal gastric cancer, LVI was more frequently observed in MSI-H than in MSS tumors (38.9% vs. 25.0%, P=0.027), but there was no difference in the presence of LN metastases. The prognosis of MSI-H tumors was similar to that of MSS tumors (log-rank test, P=0.797, the hazard ratio for MSI-H was adjusted by age, sex, pT stage, and the number of metastatic LNs, 0.932; 95% confidence interval, 0.423–2.054; P=0.861). Conclusions: MSI status was not useful in predicting prognosis in early gastric cancer. However, the frequent presence of LVI in early MSI-H gastric cancer may help guide the appropriate treatment for patients, such as endoscopic treatment or limited LN surgical dissection.

한국인의 간암에서 형광-PCR법을 이용한 Microsatellite Instability의 분석

박택규,정유진 建國大學校基礎科學硏究所 1999 理學論集 Vol.24 No.-

원발성 간암 (Hepatocellular Carcinoma; HCC)은 전세계적으로 가장 빈번하게 발생하는 암중 하나로 특히 우리나라를 비롯한 동아시아와 아프리카에서 가장 발생빈도가 높다. 이 논문에서는 이러한 간암의 분자적 수준에서의 세포의 자살기전(apoptosis)에 관련된 유전자들(wafl, Rb, p53, bcl-2, bax)과 microsatellite instability(MSI)의 상관관계를 알아보기 위하여, 12명의 감암환자 조직을 대상으로 형광 primer를 사용한 PCR을 시행하였다. 그중 wafl은 가장 높은 비율의 MSI를 나타내었으며(41.7%), 다음은 p53(25.0%)의 순이었다. 또한 전체 분석대상 검체중 85%(7/12)에서 적어도 한 개 이상의 MSI를 나타내었다. 이러한 결과는 HCC에서의 MSI의 분석이 간암의 발생과 진행에 관련되어 그 기능이 저하되거나 소멸되는 유전자들을 찾아내는데 있어서 유용한 방법이 될 수 있으며 임상적으로도 간암의 진단과 그 치료에 있어 매우 신속하고 강력한 결과를 얻기 위한 방법으로 사용될 수 있음을 시사한다. Hepatocellular carcinoma (HCC) is one of the most common cancers in many parts of the world, however the molecular mechanisms underlying liver cell transformation remain obscure. The instability of microsatellite sequences dispersed in the genome has been linked to a deficiency in cellular mismatch repair. This phenotype has been frequently observed in various human neoplasms and is regarded as a major factor in tumorigenesis. To investigate cumulative genetic changes related with apoptosis during development and progression of HCC, we examined DNAs isolated from 12 Korean HCCs and their adjacent non-tumorous parts to look for evidence of microsatellite instability (MSI). Twelve microsatellite loci (D6S271, D6S426, D13S153, D13S263, D17S849, D17S938, D17S945, D18S64, D19S420, D19S418, D19S210) were amplified by PCR form 12 Korean HCCs, and analyzed using an automated DNA analyzer. The high percentages of the MSI were found for the loci of D6S426 (33.3%) and D17S945 (25.0%). The related genes with high frequency of MSI were noted in the waf1 (41.7%) and p53 (25.0%). From this study, fifty eighty percent of HCCs (7/12) showed MSI with at least one marker. This results suggest that the analysis of MSI in HCC might be useful for identifying genes whose loss of function contributes to the development of liver cancer. Futhermore, this method may give a more rapid and accurate sizing of the PCR products of microsatellite, making the routine assessment of MSI possible in many clinical fields.

각질가시세포종에서의 이형접합성소실과 Microsatellite의 불안정성에 대한 분석

하태원(Tae-Won Ha),한기환(Ki-Hwan Han),김대광(Dae-Kwang Kim) 대한해부학회 2005 Anatomy & Cell Biology Vol.38 No.1

유전성비살버섯증잘록창자곧창자암(hereditary non-polyposis colorectal cancer)과 일부 Muir-Torre 증후군에서는 DNA부조화복구유전자 (DNA mismatch repair gene)의 돌연변이로 microsatellite 불안정성(microsatellite instability, MIS)이라는 유전자의 불안정성이 나타난다. 각질가시세포종은 형태학적으로 피부의 편평세포암종과 유사하지만 수개월간의 빠른 성장이후 완전히 소실되는 양성 종양이다. Muir-Torre 증후군 환자에서 발생한 각질가시세포종에서 MIS가 발견되어 각질가시세포종의 발생에 DNA부조화복구유전자의 이상이 중요한 역할을 할 것으로 추정되었다. 산발성각질가시세포종의 발병기전으로서 MIS와 이형접합성소실(loss of heterozygosity, LOH)의 관여 여부를 알아보기 위하여 10명의 산발성각질가시세포종 환자를 대상으로 11개의 microsatellite 표지 (D2S286, D2S367, D3S1317, D5S346, D9S160, D9S171, D10S89, D10S185, D11S904, D17S261 및 D17S520)에서 MIS과 LOH를 조사해 보았다. MIS과 LOH는 D17S261과 D10S185 표지에서 각각 1례씩 발견되었다. 결론적으로 산발성각질가시세포종에서 MIS과 LOH는 각각 10% (1/10)의 낮은 빈도로 나타나서 MIS와 LOH는 산발성각질가시세포종의 발병기전으로 큰 역할을 하지 않는 것으로 생각된다. Tumors from patients with hereditary non-polyposis colorectal cancer (HNPCC) and from a subset of patients with the related Muir-Torre syndrome exhibit a type of a genetic instability, known as microsatellite instability (MIS), which results from mutations that inactivate DNA mismatch repair genes. Keratoacanthomas resemble squamous cell carcinoma but after a period of rapid growth over a few months they involute completely. The detection of MIS in a keratoacanthoma from a patient with Muir-Torre syndrome suggested that defective mismatch repair genes may play a role in the pathogenesis of these neoplasmas. In order to elucidate the significance of both MIS and loss of heterozygosity (LOH) in the pathogenesis of sporadic keratoacanthomas, the presents of MIS and LOH at 11 microsatellite markers (D2S286, D2S367, D3S1317, D5S346, D9S16, D9S171, D10S89, D10S185, D11S904, D17S261, and D17S520) were evaluated in randomly selected sporadic keratoacanthomas. MIS and LOH were found only in 1 of 10 cases at D17S261 and D10S185, respectively. In conclusion, the low frequency of MIS and LOH detected in this study suggests that neither MIS nor LOH appear to be significant in the induction of sporadic keratoacanthomas.

위암에서 Microsatellite Instability와 Thymidylate Synthase의 상관관계

고현석,안창욱,강혜윤,김광일,홍성표,안대호 대한위암학회 2008 Journal of gastric cancer Vol.8 No.4

목적: 대장암에서 microsatellite instability high (MSI-H)를 보인 환자가 microsatellite stable (MSS) 또는 microsatellite instability low (MSI-L)를 가진 그룹보다 예후가 좋은 것으로 되어 있 으나 II기, III기 대장암에서 MSI-H를 보인 환자가 MSS 또는 MSI-L를 가진 그룹보다 5-fluorouracil (5-FU)에 대한 효과가 떨 어진다는 연구 보고가 있다. Thymidylate synthase (TS)는 DNA 합성에 필요한 물질임과 동시에 5-FU의 표적물질이며 암 환자에서 TS 발현율이 높을수록 5-FU에 의한 항암치료 효 과가 감소한다. 이와 같이 MSI가 높을수록, TS 발현이 높 을수록 5-FU에 대한 감수성이 떨어지기 때문에 MSI와 TS간 의 상관관계를 밝히려는 연구가 대장암 환자를 대상으로 시도되었으나 현재까지의 결과는 상관관계가 있다는 보고 와 없다는 보고가 있어서 일정하지 않다. 위암 환자에서는 MSI와 TS의 관계에 대한 연구는 없다. 따라서 본 연구에서 는 위암 환자에서의 MSI와 TS 발현정도의 상관관계를 분 석하였다. 대상 및 방법: 2004년 1월부터 2006년 5월까지 분당차병원 에서 위암으로 근치적 위절제술을 시행 받은 환자 중 99 명을 대상으로 MSI 및 TS 발현 정도를 비교 분석하였다. MSI는 5개의 표지자(BAT25, BAT26, D2S123, D5S346, D17S250) 에 대해서 분석하였고 TS는 면역조직화학 염색으로 그 발 현 정도를 측정하였다. 결과: 전체 99예의 환자에서 MSS/MSI-L 및 MSI-H인 경우가 각각 92 (92.9%), 7 (7.1%)예였고 TS에 대한 면역조직화학 염색 정도에 따라 negative, low TS 및 high TS인 경우는 각 각 46 (46.5%), 33 (33.3%), 20 (20.2%)예였다. MSS/MSI-L 92예에 서 TS의 negative, low TS, high TS는 각각 46 (50%), 30 (32.6%), 16 (17.4%)예였고 MSI-H인 7예에서는 TS의 negative, low TS, high TS가 0 (0%), 3 (42.9%), 4 (57.1%)예로 MSI-H 7예 모두에 서 TS를 발현하였고 검정 결과 통계적으로 유의하게 MSI-H 와 high TS 간에는 상관관계가 있었다. 결론: 위암환자에서 MSI-H를 보인 경우가 MSS/MSI-L를 보 인 경우보다 더 높은 TS의 발현을 보였다.