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

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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

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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
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