Email Alert | RSS

Journal of Peking University (Health Sciences) ›› 2023, Vol. 55 ›› Issue (2): 254-261. doi: 10.19723/j.issn.1671-167X.2023.02.008

Previous Articles     Next Articles

Analysis of microsatellite instability in endometrial cancer: The significance of minimal microsatellite shift

Li LIANG,Xin LI,Lin NONG,Ying DONG,Ji-xin ZHANG,Dong LI,Ting LI*()   

  1. Department of Pathology, Peking University First Hospital, Beijing 100034, China
  • Received:2022-09-29 Online:2023-04-18 Published:2023-04-12
  • Contact: Ting LI E-mail:lixiaoting12@hotmail.com

RICH HTML

   

PDF (PC)

Abstract:

Objective: To analyze the differences and characteristics of microsatellite instability (MSI) in endometrial cancer (EMC), by using colorectal cancer (CRC) as control. Methods: In the study, 228 cases of EMC were collected. For comparative analysis, 770 cases of CRC were collected. Mismatch repair (MMR) expression was detected by immunohistochemistry (IHC), and microsatellite instability (MSI) was analyzed by PCR and capillary electrophoresis fragment analysis (MSI-PCR). MSI-PCR was detected using five mononucleotide repeat markers: BAT-25, BAT-26, NR-21, NR-24, and MONO-27. Results: In EMC, we found 27.19% (62/228) of deficient mismatch repair (dMMR) using IHC, significantly higher than CRC (7.79%, 60/770). Meanwhile, subclonal expression of MMR protein was found in 4 cases of dMMR-EMC and 2 cases of dMMR-CRC. According to the criteria of major micro-satellite shift, we found 16.23% (37/228) of MSI-high (MSI-H), 2.63% (6/228) of MSI-low (MSI-L), and 81.14% (185/228) of microsatellite stability (MSS) in EMC using MSI-PCR. The discor-dance rate between MMR-IHC and MSI-PCR in EMC was 11.84% (27/228). In CRC, we found 8.05% (62/770) of MSI-H, 0.13% (1/770) of MSI-L, and 91.82% (707/770) of MSS. The discordance rate between MMR-IHC and MSI-PCR in CRC was only 0.52% (4/770). However, according to the criteria of minimal microsatellite shift, 12 cases of EMC showed minimal microsatellite shift including 8 cases of dMMR/MSS and 4 cases of dMMR/MSI-L and these cases were ultimately evaluated as dMMR/MSI-H. Then, 21.49% (49/228) of EMC showed MSI-H and the discordance rate MMR-IHC and MSI-PCR in EMC decreased to 6.58% (15/228). No minimal microsatellite shift was found in CRC. Compared with EMC group with major microsatellite shift, cases with minimal microsatellite shift showed younger age, better tumor differentiation, and earlier International Federation of Gynecology and Obstetrics (FIGO) stage. There were significant differences in histological variant and FIGO stage between the two groups (P < 0.001, P=0.006). Conclusion: EMC was more prone to minimal microsatellite shift, which should not be ignored in the interpretation of MSI-PCR results. The combined detection of MMR-IHC and MSI-PCR is the most sensitive and specific method to capture MSI tumors.

Key words: Endometrial cancer, Microsatellite instability, Biomarkers, tumor, Colorectal neoplasms

CLC Number: 

  • R737.33

Table 1

Comparison of consistency of MMR-IHC and MSI-PCR results between EMC and CRC"

Items EMC (n=228) CRC (n=770)
Age/years, range (media) 27-89 (56) 24-94 (63)
Gender
    Male 0 508 (65.97%)
    Female 228 (100.00%) 262 (34.03%)
MMR-IHC 62 (27.19%) 60 (7.79%)
    MLH1 (-) 1 (0.44%) 0
    PMS2 (-) 3 (1.32%) 3 (0.39%)
    MSH2 (-) 4 (1.75%) 2 (0.26%)
    MSH6 (-) 9 (3.95%) 5 (0.65%)
    MLH1/PMS2 (-) 40 (17.54%) 41 (5.32%)
    MSH2/MSH6 (-) 5 (2.19%) 9 (1.17%)
MSI-PCR (major microsatellite shift)
    MSI-H 37 (16.23%) 62 (8.05%)
    MSI-L 6 (9.68%) 1 (0.13%)
    MSS 185 (81.14%) 707 (91.82%)
MSI-PCR (minimal microsatellite shift)
    MSI-H 49 (21.49%)
    MSI-L 2 (0.88%)
    MSS 177 (77.63%)
Discordance (major microsatellite shift) 27 (11.84%) 4 (0.52%)
Discordance (minimal microsatellite shift) 15 (6.58%) 4 (0.52%)

Figure 1

Two cases of dMMR-EMC and dMMR-CRC Both cases showed the loss expression of MLH1/PMS2 and the intact expression of MSH2/MSH6; Internal positive control was detected (HE ×200, IHC ×200). EMC, endometrial cancer; CRC, colorectal cancer; dMMR, deficient mismatch repair; HE, hematoxylin-eosin; IHC, immunohistochemistry."

Figure 2

Subclone expression of MMR-IHC The positive expression of MLH1 and PMS2 were detected in some tumor cells, while no positive expression was found in the other region in one case of EMC. Similarly, the subclone expression of MSH6 was found in one case of CRC. Internal positive control was detected (IHC ×200). Abbreviations as in Figure 1."

Figure 3

Representative results of MSI-PCR A, a representative result of MSI-PCR from dMMR-EMC with the discordance between MMR-IHC and MSI-PCR. Compared with its paired normal control, tumor group showed minimal microsatellite shift (+2 bp) in the mononucleotide site BAT-26, BAT-25 and NR-21 (red arrow). B, a representative result of MSI-PCR from dMMR-EMC with the consistency between MMR-IHC and MSI-PCR. Compared with its paired normal control, tumor group showed major microsatellite shift (+3-5 bp) in the mononucleotide site BAT-25 and MONO-27. C, a representative result of MSI-PCR from dMMR/MSI-H CRC. Compared with its paired normal control, tumor group showed major microsatellite shift (+5-8 bp, major peak) in the mononucleotide site BAT-25 and MONO-27. The title of the ordinate axis is relative fluorescence units. The title of the abscissa axis is fragment size (bp). EMC, endometrial cancer; CRC, colorectal cancer; dMMR, deficient mismatch repair; MMR, mismatch repair; IHC, immunohistochemistry; MSI, microsatellite instability; MSI-H, MSI-high."

Figure 4

Frequency of minimal microsatellite shift in five mononucleotide sites (BAT-26, NR-21, BAT-25, MONO-27, and NR-24)"

Figure 5

Profile of MMR-IHC and MSI-PCR results from 228 cases of endometrial cancer EMC, endometrial cancer; dMMR, deficient mismatch repair; pMMR, proficient mismatch repair; IHC, immunohistochemistry; MSI, microsatellite instability; MSI-H, MSI-high; MSI-L, MSI-low; MSS, microsatellite stability."

Table 2

Comparison between dMMR/minimal microsatellite shift group and dMMR/major microsatellite shift group in endometrial cancer"

Items dMMR/minimal microsatellite shift
(n=12)		 dMMR/major microsatellite shift
(n=36)
Age/years, range (media) 48-68 (55) 41-89 (57)
Histology
    Well differentiated EMC 5 (41.67%) 2 (5.56%)
    Moderately differentiated EMC 4 (33.33%) 17 (47.22%)
    Poorly differentiated EMC 3 (25.00%) 11 (30.56%)
    Serous carcinoma 0 1 (2.78%)
    Clear cell carcinoma 0 1 (2.78%)
    Mixed carcinoma 0 0
    Dedifferentiated carcinoma 0 1 (2.78%)
    Undifferentiated carcinoma 0 1 (2.78%)
    Carcinosarcoma 0 2 (5.56%)
FIGO stage
    ⅠA 11 (91.67%) 20 (55.56%)
    ⅠB 1 (8.33%) 10 (27.78%)
    Ⅱ 0 4 (11.11%)
    ⅢC1 0 1 (2.78%)
    ⅢC2 0 1 (2.78%)
dMMR
    MLH1 (-) 0 1 (2.78%)
    PMS2 (-) 1 (8.33%) 0
    MSH2 (-) 0 4 (11.11%)
    MSH6 (-) 2 (16.67%) 2 (5.56%)
    MLH1/PMS2 (-) 9 (75.00%) 24 (66.67%)
    MSH2/MSH6 (-) 0 5 (13.89%)

Table 3

Comparison of histology and FIGO stage between dMMR/minimal microsatellite shift group and dMMR/major microsatellite shift group in endometrial cancer"

Items dMMR/minimal microsatellite shift dMMR/major microsatellite shift P
Histology, n
    Well differentiated group 11 2 < 0.001
    Poorly differentiated group 15 34
FIGO stage, n
    ⅠA 23 20 0.006
    ⅠB/Ⅱ/ⅢC/ⅢC2 3 16
1 Yamamoto H , Imai K . Microsatellite instability: An update[J]. Arch Toxicol, 2015, 89 (6): 899- 921.
doi: 10.1007/s00204-015-1474-0
2 Baretti M , Le DT . DNA mismatch repair in cancer[J]. Pharmacol Ther, 2018, 189, 45- 62.
doi: 10.1016/j.pharmthera.2018.04.004
3 Lynch HT , Shaw MW , Magnuson CW , et al. Hereditary factors in cancer. Study of two large midwestern kindreds[J]. Arch Intern Med, 1966, 117 (2): 206- 212.
doi: 10.1001/archinte.1966.03870080050009
4 Dudley JC , Lin MT , Le DT , et al. Microsatellite instability as a biomarker for PD-1 blockade[J]. Clin Cancer Res, 2016, 22 (4): 813- 820.
doi: 10.1158/1078-0432.CCR-15-1678
5 Kandoth C , Schultz N , Cherniack AD , et al. Integrated genomic characterization of endometrial carcinoma[J]. Nature, 2013, 497 (7447): 67- 73.
doi: 10.1038/nature12113
6 Benson AB , Venook AP , Al-Hawary MM , et al. Colon cancer, version 2.2021, NCCN clinical practice guidelines in oncology[J]. J Natl Compr Canc Netw, 2021, 19 (3): 329- 359.
doi: 10.6004/jnccn.2021.0012
7 Koh WJ , Abu-Rustum NR , Bean S , et al. Uterine neoplasms, version 1.2018, NCCN clinical practice guidelines in oncology[J]. J Natl Compr Canc Netw, 2018, 16 (2): 170- 199.
doi: 10.6004/jnccn.2018.0006
8 Libera L , Sahnane N , Pepe F , et al. Critical aspects of micro-satellite instability testing in endometrial cancer: A comparison study[J]. Hum Pathol, 2022, 128, 134- 140.
doi: 10.1016/j.humpath.2022.07.014
9 Tafe LJ , Riggs ER , Tsongalis GJ . Lynch syndrome presenting as endometrial cancer[J]. Clin Chem, 2014, 60 (1): 111- 121.
doi: 10.1373/clinchem.2013.206888
10 Recommendations from the EGAPP Working Group . Genetic testing strategies in newly diagnosed individuals with colorectal cancer aimed at reducing morbidity and mortality from Lynch syndrome in relatives[J]. Genet Med, 2009, 11 (1): 35- 41.
doi: 10.1097/GIM.0b013e31818fa2ff
11 Wu X , Snir O , Rottmann D , et al. Minimal microsatellite shift in microsatellite instability high endometrial cancer: A significant pitfall in diagnostic interpretation[J]. Mod Pathol, 2019, 32 (5): 650- 658.
doi: 10.1038/s41379-018-0179-3
12 Wang Y , Shi C , Eisenberg R , et al. Differences in microsatellite instability profiles between endometrioid and colorectal cancers: A potential cause for false-negative results?[J]. J Mol Diagn, 2017, 19 (1): 57- 64.
doi: 10.1016/j.jmoldx.2016.07.008
13 Longacre TA , Broaddus R , Chuang LT , et al. Template for reporting results of biomarker testing of specimens from patients with carcinoma of the endometrium[J]. Arch Pathol Lab Med, 2017, 141 (11): 1508- 1512.
doi: 10.5858/arpa.2016-0450-CP
14 Cortes-Ciriano I , Lee S , Park WY , et al. A molecular portrait of microsatellite instability across multiple cancers[J]. Nat Commun, 2017, 8, 15180.
doi: 10.1038/ncomms15180
15 Guyot D'Asnières , De Salins A , Tachon G , Cohen R , et al. Discordance between immunochemistry of mismatch repair proteins and molecular testing of microsatellite instability in colorectal cancer[J]. ESMO Open, 2021, 6 (3): 100120.
doi: 10.1016/j.esmoop.2021.100120
16 Ferguson SE , Aronson M , Pollett A , et al. Performance characte-ristics of screening strategies for Lynch syndrome in unselected women with newly diagnosed endometrial cancer who have under-gone universal germline mutation testing[J]. Cancer, 2014, 120 (24): 3932- 3939.
doi: 10.1002/cncr.28933
17 McConechy MK , Talhouk A , Li-Chang HH , et al. Detection of DNA mismatch repair (MMR) deficiencies by immunohistoche-mistry can effectively diagnose the microsatellite instability (MSI) phenotype in endometrial carcinomas[J]. Gynecol Oncol, 2014, 137 (2): 306- 310.
18 Marshman E , Booth C , Potten CS . The intestinal epithelial stem cell[J]. Bioessays, 2002, 24 (1): 91- 98.
doi: 10.1002/bies.10028
19 Gargett CE , Nguyen HP , Ye L . Endometrial regeneration and endometrial stem/progenitor cells[J]. Rev Endocr Metab Disord, 2012, 13 (4): 235- 251.
doi: 10.1007/s11154-012-9221-9
20 Metcalf AM , Spurdle AB . Endometrial tumour BRAF mutations and MLH1 promoter methylation as predictors of germline mismatch repair gene mutation status: A literature review[J]. Fam Cancer, 2014, 13 (1): 1- 12.
doi: 10.1007/s10689-013-9671-6
21 Schweizer P , Moisio AL , Kuismanen SA , et al. Lack of MSH2 and MSH6 characterizes endometrial but not colon carcinomas in hereditary nonpolyposis colorectal cancer[J]. Cancer Res, 2001, 61 (7): 2813- 2815.
22 Hampel H , Panescu J , Lockman J , et al. Comment on: Screening for Lynch syndrome (hereditary nonpolyposis colorectal cancer) among endometrial cancer patients[J]. Cancer Res, 2007, 67 (19): 9603.
doi: 10.1158/0008-5472.CAN-07-2308
23 Zhang L . Immunohistochemistry versus microsatellite instability testing for screening colorectal cancer patients at risk for hereditary nonpolyposis colorectal cancer syndrome. Part Ⅱ. The utility of microsatellite instability testing[J]. J Mol Diagn, 2008, 10 (4): 301- 307.
doi: 10.2353/jmoldx.2008.080062
24 Stelloo E , Jansen AML , Osse EM , et al. Practical guidance for mismatch repair-deficiency testing in endometrial cancer[J]. Ann Oncol, 2017, 28 (1): 96- 102.
doi: 10.1093/annonc/mdw542
25 Ta RM , Hecht JL , Lin DI . Discordant loss of mismatch repair proteins in advanced endometrial endometrioid carcinoma compared to paired primary uterine tumors[J]. Gynecol Oncol, 2018, 151 (3): 401- 406.
doi: 10.1016/j.ygyno.2018.10.012
[1] Dan HAN, Yangjin CIREN, Qiuhong LI, Jun LI. Changes of intestinal microflora in patients with colorectal benign and malignant tumors in high altitude area and comparison with the normal population in low altitude area [J]. Journal of Peking University (Health Sciences), 2025, 57(3): 578-583.
[2] DING Ting-ting,ZENG Chu-xiong,HU Li-na,YU Ming-hua. Establishment of a prediction model for colorectal cancer immune cell infiltration based on the cancer genome atlas (TCGA) database [J]. Journal of Peking University (Health Sciences), 2022, 54(2): 203-208.
[3] Xu-chu ZHANG,Jian-hua ZHANG,Rong-fu WANG,Yan FAN,Zhan-li FU,Ping YAN,Guang-yu ZHAO,Yan-xia BAI. Diagnostic value of 18F-FDG PET/CT and tumor markers (CEA, CA19-9, CA24-2) in recurrence and metastasis of postoperative colorectal moderately differentiated adenocarcinoma [J]. Journal of Peking University(Health Sciences), 2019, 51(6): 1071-1077.
[4] WANG Xiao-hui， ZHANG Yan， LIU Lin-zhi， SHANG Chen-guang . Effects of metformin and adiponectin on endometrial cancer cells growth [J]. Journal of Peking University(Health Sciences), 2018, 50(5): 767-773.
[5] CHEN Yang, WANG Yan-rong, SHI Yan, DAI Guang-hai. Prognostic value of chemotherapy-induced neutropenia in metastatic colon cancer patients undergoing first-line chemotherapy with FOLFOX [J]. Journal of Peking University(Health Sciences), 2017, 49(4): 669-674.
[6] LIU Yan-xia, YANG Xue-song, FU Wei, YAO Hong-wei. Association of single nucleotide polymorphism rs6983267 with ulcerative colitis and colorectal cancer [J]. Journal of Peking University(Health Sciences), 2016, 48(6): 994-999.
[7] SI Jing-wen, WANG Li, BA Xiao-jun, ZHANG Xu, DONG Ying, ZHANG Ji-xin, LI Wen-ting, LI Ting. Clinicopathological screening of Lynch syndrome: a report of 2 cases and literature review [J]. Journal of Peking University(Health Sciences), 2015, 47(5): 858-864.
[8] YUAN Chun-hui, XIU Dian-rong, GE Hui-yu, TAN Shi, WANG Hang-yan, ZHANG Li, ZHANG Tong-lin. Ultrasound guided ablation therapy of hepatic colorectal metastases: initial experience of real time virtual sonography navigation system [J]. Journal of Peking University(Health Sciences), 2013, 45(6): 956-959.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright © Journal of Peking University (Health Sciences), All Rights Reserved.
Powered by Beijing Magtech Co. Ltd