外科理论与实践 ›› 2024, Vol. 29 ›› Issue (02): 148-155.doi: 10.16139/j.1007-9610.2024.02.10
朱惠a, 蔡继东b,*, 李溟涵c, 杨文涛c(
), 徐烨c
收稿日期:2024-02-24
出版日期:2024-03-25
发布日期:2024-07-01
通讯作者:
杨文涛，E-mail: yangwt97@163.com作者简介:共同第一作者
ZHU Huia, CAI Jidongb,*, LI Minghanc, YANG Wentaoc(), XU Yec
Received:2024-02-24
Online:2024-03-25
Published:2024-07-01
摘要：
目的：探究免疫组织化学(IHC)检测错配修复(MMR)状态与二代测序(NGS)鉴定的微卫星不稳定性(MSI)的一致性，并评估其结果与中国结肠直肠癌（CRC）病人临床特征的相关性。方法：采用IHC和NGS测序分别鉴定CRC的MMR及MSI状态，评估不同检测方式的一致性。结果：IHC与NGS检测方式一致性为98.36%，一致性良好（Kappa=0.856）。pMMR/MSI-H亚型中存在一定的致病性或可能致病性胚系变异，dMMR/MSS亚型中MLH1和PMS2共同缺失最为常见。分型不一致的病人多集中于发病较早的右半结肠（P<0.01），且分化相对较差。结论：经IHC和NGS检测的MSI一致性很好，可高达98%以上。为避免因MSI状态误诊而影响临床医师对治疗方案的决策，对于分化较差的早期阶段右半结肠必须高度重视MSI分析的准确性。
中图分类号:
朱惠, 蔡继东, 李溟涵, 杨文涛, 徐烨. 结肠直肠癌错配修复蛋白表达与微卫星稳定性的一致性分析[J]. 外科理论与实践, 2024, 29(02): 148-155.
ZHU Hui, CAI Jidong, LI Minghan, YANG Wentao, XU Ye. Consistency analysis of mismatch repair protein expression and microsatellite stability in colorectal cancer[J]. Journal of Surgery Concepts & Practice, 2024, 29(02): 148-155.
表1
采用IHC和NGS检测的微卫星状态的CRC的临床特征[n(%)/M(Q1~Q3)]
| Clinical Characteristics | 总计 (n=2 196) | IHC | P value | NGS | P value | ||||
|---|---|---|---|---|---|---|---|---|---|
| pMMR (n=2 069) | dMMR(n=127) | χ2 value | MSI-H (n=139) | MSS (n=2 057) | χ2 value | ||||
| Gender | |||||||||
| Male | 1 313 (59.79) | 1 234 (59.64) | 79(62.20) | 0.326 8 | 0.567 6 | 83 (59.71) | 1 230 (59.80) | 0.000 4 | 0.984 5 |
| Female | 883 (40.21) | 835 (40.36) | 48 (37.80) | 56 (40.29) | 827 (40.20) | ||||
| Age | |||||||||
| mAge（range） | 61 (20-93) | 61 (21-93) | 55 (20.88) | 55 (20-88) | 61 (21-93) | ||||
| ≤60 | 1 075 (48.95) | 999 (48.28) | 76 (59.84) | 6.396 9 | 0.011 | 82 (58.99) | 993 (48.27) | 5.986 1 | 0.014 4 |
| >60 | 1 121 (51.05) | 1 070 (51.72) | 51 (40.16) | 57 (41.01) | 1 064 (51.73) | ||||
| Primary site | |||||||||
| Left | 1 625 (74.00) | 1 583 (76.51) | 42 (33.07) | 117.348 0 | <0.01 | 47 (33.81) | 1 578 (76.71) | 124.543 0 | <0.01 |
| Right | 571 (26.00) | 486 (23.49) | 85 (66.93) | 92 (66.19) | 479 (23.29) | ||||
| AJCC | |||||||||
| Ⅰ-Ⅲ | 1 645 (74.91) | 1 529 (73.90) | 116 (91.34) | 19.358 9 | <0.01 | 125 (89.93) | 1 520 (73.89) | 17.804 9 | <0.01 |
| Ⅳ | 551 (25.09) | 540 (26.10) | 11 (8.66) | 14 (10.07) | 537 (26.11) | ||||
| Differentiation | |||||||||
| Well | 39 (1.78) | 36 (1.74) | 3 (2.36) | 19.070 3 | <0.01 | 1 (0.72) | 38 (1.85) | 19.160 6 | <0.01 |
| Medium | 1 476 (67.21) | 1 413 (68.29) | 63 (49.61) | 72 (51.80) | 1 404 (68.25) | ||||
| Low | 681 (31.01) | 620 (29.97) | 61 (48.03) | 66 (47.48) | 615 (29.90) | ||||
表2
IHC检测的MMR蛋白表达情况[n (%)]
| MMR | MLH1 | MSH2 | MSH6 | PMS2 | n (%) |
|---|---|---|---|---|---|
| Expression of MMR proteins | （-） | （+） | （+） | （-） | 77 (60.63) |
| （+） | （-） | （-） | （+） | 23 (18.11) | |
| （+） | （+） | （+） | （-） | 15 (11.81) | |
| （+） | （-） | （+） | （+） | 4 (3.15) | |
| （+） | （+） | （-） | （+） | 3 (2.36) | |
| （+） | （+） | （-） | （-） | 2 (1.57) | |
| （-） | （+） | （+） | （+） | 1 (0.79) | |
| （-） | （+） | （-） | （-） | 1 (0.79) |
表3
NGS检测的MSI-H亚组MMR基因胚系突变情况[n (%)]
| Type of causation | Pathogenic variants | Likely pathogenic variants | |||
|---|---|---|---|---|---|
| MMR | n (%) | Type of variation | n (%) | Type of variation | |
| MLH1 | 9 (26.47) | Missense mutation×3, non-frameshift deletion×2, Frame shift missing×1, frame shift insertion×1，splice×1, intron×1 | 6 (17.65) | Frameshift missing×3, complex mutation×1，missense mutation×1，splice×1 | |
| MSH2 | 4 (11.76) | Frameshift missing×3 Splice×1 | 5 (14.71) | Frameshift insertion×2 Frameshift missing×3 | |
| MSH6 | 3 (8.82) | Stop×2 Frame shift insertion×1 | 1 (2.94) | Frame shift insertion×1 | |
| PMS2 | 1 (2.94) | Stop×1 | 5 (14.71) | Frameshift missing×2, Stop×1, Frameshift insertion×1 | |
| Total | 17 (50.00) | 17 (50.00) | |||
表4
IHC与NGS检测的微卫星状态的差异性[n (%)]
| MSI-status | MSI-H | MSS | Total |
|---|---|---|---|
| pMMR | 24 (1.09) | 2 045 (93.12) | 2 069 |
| dMMR | 115 (5.24) | 12 (0.55) | 127 |
| Total | 139 | 2 057 | 2 196 |
表5
不一致的微卫星状态下IHC中MMR蛋白表达情况
| MMR | MLH1 | MSH2 | MSH6 | PMS2 | n (%) | MMR_ status | MSI_ status |
|---|---|---|---|---|---|---|---|
| Experssion | （+） | （+） | （+） | （+） | 24 (66.67) | pMMR | MSI-H |
| （-） | （+） | （+） | （-） | 6 (16.67) | dMMR | MSS | |
| （+） | （+） | （+） | （-） | 3 (8.33) | dMMR | MSS | |
| （+） | （-） | （-） | （+） | 2 (5.56) | dMMR | MSS | |
| （+） | （+） | （-） | （+） | 1 (2.78) | dMMR | MSS |
表6
不一致的微卫星状态下NGS测序中MMR基因胚系变异情况
| Case(n) | Germline mutations | MMR status | MSI status | Gene | Type of variation | Type of causation |
|---|---|---|---|---|---|---|
| 19 | No | pMMR | MSI-H | - | - | - |
| 1 | Yes | pMMR | MSI-H | PMS2 | Stop | Pathogenic |
| 1 | Yes | pMMR | MSI-H | MSH6 | Frame shift insertion | Pathogenic |
| 1 | Yes | pMMR | MSI-H | MSH6 | Frame shift insertion | Likely |
| 1 | Yes | pMMR | MSI-H | MLH1 | Frame shift missing | Pathogenic |
| 1 | Yes | pMMR | MSI-H | PMS2 | Frame shift insertion | Likely |
表7
微卫星状态不一致的CRC临床特征[n (%)/M(Q1~Q3)]
| Clinical Characteristics | Total (n=2 196) | Consistent （n=2 160） | Inconsistent （n=36） | χ2 value | P value |
|---|---|---|---|---|---|
| Gender | |||||
| Male | 1 313 (59.79) | 1 293 (59.86) | 20 (55.56) | 0.273 0 | 0.601 3 |
| Female | 883 (40.21) | 867 (40.14) | 16 (44.44) | ||
| Age | |||||
| mAge（range） | 61 (20.93) | 63 (20-93) | 63 (29-80) | ||
| ≤60 | 1 075 (48.95) | 1 061 (49.12) | 14 (38.89) | 1.483 3 | 0.223 2 |
| >60 | 1 121 (51.05) | 1 099 (50.88) | 22 (61.11) | ||
| Primary site | |||||
| Left | 1 625 (74.00) | 1 608 (74.44) | 17 (47.22) | 13.637 9 | <0.01 |
| Right | 571 (26.00) | 552 (25.56) | 19 (52.78) | ||
| AJCC | |||||
| Ⅰ-Ⅲ | 1 645 (74.91) | 1 616 (74.81) | 29 (80.56) | 0.620 9 | 0.430 7 |
| Ⅳ | 551 (25.09) | 544 (25.19) | 7 (19.44) | ||
| Differentiation | |||||
| Well | 39 (1.78) | 37 (1.71) | 2 (5.56) | 3.680 3 | 0.158 8 |
| Medium | 1 476 (67.21) | 1 455 (67.36) | 21 (58.33) | ||
| Low | 681 (31.01) | 668 (30.93) | 13 (36.11) |
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