原发、复发及转移胃肠道间质瘤KIT及PDGFRA突变与临床病理特征间的关系

doi:10.16150/j.1671-2870.2021.03.006

摘要/Abstract

摘要：

目的:探讨原发、复发及转移性胃肠道间质瘤（gastrointestinal stromal tumor,GIST）中的KIT及PDGFRA基因突变与免疫表型、临床病理学特征之间的相关性。方法:收集2004年到2020年本院167例资料完整的原发GIST患者,其中复发及转移患者22例。运用免疫组织化学（免疫组化）染色及第一代测序技术对167例原发GIST病灶及22例复发或转移转移灶进行CD117、DOG1、CD34和Ki-67增殖指数检测,以及KIT基因第9、11、13、17号外显子和PDGFRA基因第12、18号外显子测序。结果:167例原发GIST中KIT和PDGFRA基因的突变率分别为83.8%（140/167）和3.0%（5/167）。KIT外显子11突变率为74.9%（125/167）,外显子9突变率为8.4%（14/167）,外显子17突变率为0.6%（1/167）。5例GIST原发灶中存在PDGFRA基因外显子18突变,其中4例为D842V突变。KIT外显子突变类型包括点突变（34.3%,48/140）、缺失突变（40.7%,57/140）、重复突变（3.6%,5/140）、混合突变（插入合并缺失,12.1%,17/140）,其中携带KIT外显子11非点突变的GIST（n=76）Ki-67增殖指数（10.7%）高于携带外显子11点突变（n=45）的GIST（8.1%）（P=0.005 2）。22例复发及转移GIST病灶中,KIT双外显子突变共有9例,其中6例为外显子11和17双突变,2例为外显子11和13双突变,1例为外显子9和13双突变。在22例复发、转移GIST患者中,与KIT单突变（中位时间60个月）及野生型（中位时间30个月）相比,KIT双外显子突变患者的无疾病进展时间显著延长（中位时间为108个月,P=0.029 9和P=0.011 1）,而Ki-67表达值、CD117、DOG1、CD34、肿瘤直径及核分裂象与前二者间差异无统计学意义。结论:原发GIST灶中,以KIT基因外显子11突变为主,缺失突变最多,占40.7%;原发灶KIT基因外显子11非点突变的肿瘤患者,其Ki-67表达值高于点突变类型患者,可能提示该类患者的肿瘤生物学行为更差。与原发灶KIT基因突变模式均为单外显子突变不同,复发及转移灶存在KIT基因双外显子突变,且占比较高（9/22）。在肿瘤转移、复发患者中,与野生型及单突变者相比,KIT基因双外显子突变者无疾病进展时间较长。

关键词: 胃肠道间质瘤, KIT基因, PDGFRA基因, 临床病理

Abstract:

Objective: To investigate the correlation of KIT and PDGFRA mutation status with immunohistochemistry (IHC) and clinicopathological features in the primary, recurren tor metastatic gastrointestinal stromal tumors (GIST). Methods: A total of 167 primary GIST including 22 recurrent or metastatic GIST were tested for CD117, DOG1, CD34 and Ki-67 by IHC, and KIT gene exon 9, 11, 13, 17and PDGFRA gene exon12, 18 were analyzed by Sanger sequencing. Results: The frequencies of KIT and PDGFRA mutation in 167 primary GIST were 83.8%(140/167) and 3.0% (5/167), respectively. The mutation frequencies of KIT gene exon 11, exon 9 and exon 17 were 74.9% (125/167),8.4% (14/167) and 0.6% (1/167). Five primary GIST harbored PDGFRA mutations in exon 18,and 4 tumors had D842V mutation. Genetic mutations in KIT gene included point mutations (34.3%,48/140), deletion mutations (40.7%,57/140), duplication mutations (3.6%,5/140), and complicated mutations (deletion-insertion,12.1%,17/140).For GIST with KIT gene exon 11 mutation, Ki-67 index was significantly higher in tumors with non-point mutation than those with point mutations (P=0.0052). In 22 recurrent or metastatic GIST, double-exon mutation on KIT gene were found in 9 cases and 6 case were with mutation in both exon 11 and 17, 2 in exon 11 and 13,and one in exon 9 and 13.In 22 cases with recurrent or metastatic GIST, patients with double KIT exon mutations had a longer median progression-free survival (PFS)(108 months) than those with wild type gene (PFS, 30 months),and single exon mutation (PFS, 60 months)(P=0.0299 and P=0.0111), while no significant difference were observed regarding Ki-67, CD117, DOG1, CD34 expression,tumor size and nuclear mitosis between them. Conclusions: KIT mutations in primary GIST occur more often in exon 11. For primary GIST with KIT exon 11 mutations, Ki-67 index is higher in lesions with non-point mutation than those with point mutation,indicating poor biological beha-vior. In recurrence or metastatic GIST, cases harbor the double KIT exon mutations account for 9/22 and possess a better PFS than those with wild type and singe KIT exon 11 mutation.

Key words: Gastrointestinal stromal tumors, KIT mutation, PDGFRA mutation, Clinical pathological features

中图分类号:

R735

顾怡瑾, 李安琪, 董磊, 许海敏, 申霞, 谢嘉玲, 袁菲, 王朝夫. 原发、复发及转移胃肠道间质瘤KIT及PDGFRA突变与临床病理特征间的关系[J]. 诊断学理论与实践, 2021, 20(03): 257-264.

GU Yijin, LI Anqi, DONG Lei, XU Haimin, SHEN Xia, XIE Jialing, YUAN Fei, WANG Chaofu. Correlation of KIT and PDGFRA mutation status with clinicopathologic features in primary and recurrent or metastatic gastrointestinal stromal tumors[J]. Journal of Diagnostics Concepts & Practice, 2021, 20(03): 257-264.

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抗体名称	抗体克隆号	HIER所需温度、修复液pH和浓度	一抗孵育条件	抗体来源公司
CD117	YR145	97 ℃,pH=9.0,即用型	32 ℃ 30 min	福州迈新
DOG1	OTI1C6	97 ℃,pH=9.0,即用型	32 ℃ 30 min	北京中杉
CD34	QBEnd/10	97 ℃,pH=9.0,即用型	32 ℃ 30 min	DAKO
Ki67	MIB-1	97 ℃,pH=6.0,即用型	32 ℃ 20 min	DAKO

临床病理参数	KIT 突变	PDGFRA 突变	野生型	P值
年龄(岁)				0.647
≤60	65	3	12
>60	75	2	10
性别				0.169
男	69	3	7
女	71	2	15
肿瘤部位				0.212
胃	77	4	9
小肠	36	0	9
结肠直肠	3	0	0
胃肠道外	24	1	4
肿瘤直径(cm)^a)				0.538
≤2	16	0	1
2~5^b)	42	1	5
5~10	47	3	10
≥10	23	0	5
核分裂象(50个/HPF)^a)				0.400
≤5	82	4	16
>5	31	1	3
DOG-1^a)				0.127
+	139	5	20
-	0	0	1
CD117^a)				1.000
+	139	3	21
-	0	2	0
CD34^a)				1.000
+	120	5	19
-	17	0	2
Ki-67增殖指数(%)^a)				0.183
≤5	84	5	16
5~10	3	0	0
≥10	49	0	3
危险度^a),c)				0.067
低度	51	1	5
中度	20	2	8
高度	61	1	9