胶质瘤中MGMT甲基化状态及其临床意义的回顾性研究

doi:10.19401/j.cnki.1007-3639.2023.08.003

摘要/Abstract

摘要：

背景与目的：胶质瘤是中枢神经系统常见且预后较差的恶性肿瘤，手术后联合替莫唑胺（temozolomide，TMZ）同步放化疗是胶质瘤的主要治疗方案。O6-甲基鸟嘌呤DNA甲基转移酶（O6-methylguanine DNA methyltranferase，MGMT）基因启动子甲基化（MGMT promoter methylation，MGMTmet）状态可预测胶质瘤患者对TMZ治疗的敏感性，但其与临床病理学特征的关系及如何更好地预测治疗效果及患者预后尚需深入研究。本研究旨在分析胶质瘤中MGMTmet状态及其与临床病理学特征和其他常见分子异常的相关性，探讨MGMTmet与其他分子异常联合分析对胶质瘤患者预后和TMZ治疗效果判断的价值。方法：回顾性收集复旦大学附属肿瘤医院病理科2019年7月—2022年9月诊断的205例胶质瘤患者的临床病理学资料，采用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）法检测MGMTmet状态；采用Sanger测序法检测异柠檬酸脱氢酶（isocitrate dehydrogenase，IDH）1和IDH2端粒酶反转录酶（telomerase reverse transcriptase，TERT）基因突变情况；采用荧光原位杂交（fluorescence in situ hybridization，FISH）检测1号染色体短臂和19号染色体长臂（the short arm of chromosome 1 and the long arm of chromosome 19，1p19q）缺失情况。结果：205例患者中，女性患者的MGMTmet发生率高于男性。相比于胶质母细胞瘤（47.3%），星形细胞瘤（74.1%）和少突胶质细胞瘤（100.0%）中MGMT基因启动子更易发生甲基化（P<0.05）。在MGMTmet组中，IDH1突变（mutant，mut）率和1p19q共缺失（co-deletion，co-del）率明显提高，且MGMTmet与IDH1mut和1p19qco-del具有相关性（P<0.05）。MGMTmet、年龄小于55岁、少突胶质细胞瘤及世界卫生组织（World Health Organization，WHO） 1~3级的患者均表现出较长的总生存期（overall survival，OS），差异有统计学意义（P<0.05）。相比于单个影响因素，双/三基因联合分析[MGMTmet/IDH1mut、MGMTmet/1p19q co-del或MGMTmet/IDH1mut/1p19q co-del]预测患者预后的效果更好（P<0.05），后两者是独立预后因素。在TMZ治疗患者中，MGMTmet（MGMTmet/TMZ+）患者比其他组预后好，如果患者联合存在IDH1mut，患者预后得到进一步提高（P<0.05）。结论：MGMTmet好发于女性和少突胶质细胞瘤患者中；其与IDH1mut及1p19 qco-del呈正相关。MGMTmet的患者与TMZ治疗效果及预后较好有关，且MGMTmet联合IDH突变和1p19q co-del分析可能具有更好的TMZ治疗效果和预后提示价值。

关键词: 胶质瘤, O6-甲基鸟嘌呤DNA甲基转移酶, 双/三基因联合分析, 预后, 替莫唑胺

Abstract:

Background and purpose: Glioma is a common malignant tumor of central nervous system with poor prognosis. Postoperative concurrent chemoradiotherapy with temozolomide (TMZ) is the main treatment for glioma. The methylation status of the O6-methylguanine-DNA methyltransferase (MGMT) promoter can predict the sensitivity of glioma patients to TMZ treatment, however its relationship with clinical pathology and how to better predict treatment and prognosis still need further research. The purpose of this study was to analyze the status of MGMT promoter methylation (MGMTmet) in gliomas and its correlation with clinical pathological features and other common molecular abnormalities, and to explore the value of combined analysis of MGMTmet and other molecular abnormalities in predicting the prognosis of glioma and the efficacy of TMZ treatment. Methods: We retrospectively collected clinical and pathological data from 205 glioma patients diagnosed by the Department of Pathology, Fudan University Shanghai Cancer Center from July 2019 to September 2022. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect MGMTmet status. Sanger sequencing was used to detect the mutation of isocitrate dehydrogenase 1 and 2 (IDH1/2) and telomerase reverse transcriptase (TERT) genes. Fluorescence in situ hybridization (FISH) was used to detect the deletion of the short arm of chromosome 1 and the long arm of chromosome 19 (1p19q). Results: Among 205 patients, the incidence of MGMTmet was higher in female patients than in male patients. Compared to glioblastoma (47.3%), astrocytoma (74.1%) and oligodendroglioma (100.0%) were more prone to methylation of the MGMT gene promoter (P<0.05). In MGMTmet group, IDH1 mutation rate and 1p19q co-deletion rate were significantly increased, and methylation of MGMT promoter was correlated with IDH1 mutation and 1p19q co-deletion (P<0.05). Patients with MGMTmet, age less than 55 years, oligodendroglioma, and World Health Organization (WHO) grade 1-3 all showed longer overall survival (OS), and the difference is statistically significant (P<0.05). Compared with individual influencing factors, dual/triple gene combination analysis (MGMTmet/IDH1 mutation or MGMTmet/1p19q co-deletion or MGMTmet/IDH1 mutation/1p19q co-deletion) had better effect for predicting the patient prognosis (P<0.05), with the latter two being independent prognostic factors. Among TMZ treated patients, MGMTmet (MGMTmet/TMZ+) patients had a better prognosis than other groups. If the patients had combined IDH1 mutations, the prognosis of the patients was further improved (P<0.05). Conclusion: MGMTmet is more common in women and patients with oligodendroglioma. It is positively correlated with IDH1 mutation and 1p19q co-deletion. Patients with MGMTmet are associated with better TMZ treatment efficacy and prognosis, and MGMTmet combined with IDH mutations and 1p19q co-deletion analysis have better TMZ treatment efficacy and prognostic implications.

Key words: Glioma, O6 methylguanine DNA methyltransferase, Double/triple gene conjoint analysis, Prognosis, Temozolomide

中图分类号:

R739.41

姜琳, 刘绮颖, 贾利晴, 张静, 常恒, 薛田, 任敏, 柏乾明, 朱晓丽, 周晓燕. 胶质瘤中MGMT甲基化状态及其临床意义的回顾性研究[J]. 中国癌症杂志, 2023, 33(8): 740-750.

JIANG Lin, LIU Qiying, JIA Liqing, ZHANG Jing, CHANG Heng, XUE Tian, REN Min, BAI Qianming, ZHU Xiaoli, ZHOU Xiaoyan. Retrospective study on MGMT methylation status and its clinical significance in gliomas[J]. China Oncology, 2023, 33(8): 740-750.

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Characteristic	Case n	MGMT promotor methylation	χ²	P value
Gender			4.037	0.045
Male	125	70
Female	80	56
Age/year			2.333	0.127
<55	120	79
≥55	85	47
Tumor location			7.026	0.071
Cerebellum brain stem	6	3
Temporo-occipital lobe	47	25
Frontal lobe	69	51
Others	83	47
Tumor histological type			42.080	<0.001
Glioblastoma	93	44
Oligodendroglioma	33	33
Astrocytoma	58	43
Others	21	6
WHO grade			26.730	<0.001
4	120	57
3	36	33
1+2	49	36

Molecular characteristics	MGMTmet (n=127)	MGMTunmet (n=78)	χ²	P value
IDH1mut			35.248	<0.001
-	53 (41.7)	60 (76.9)
+	67 (52.8)	9 (11.5)
NA	7 (5.5)	9 (11.5)
IDH2mut			4.391	0.111
-	111 (87.4)	69 (88.5)
+	6 (4.7)	0 (0.0)
NA	10 (7.9)	9 (11.5)
1p19q co-deletion			15.588	<0.001
-	42 (33.1)	28 (35.9)
+	36 (28.3)	5 (6.4)
NA	49 (38.6)	45 (57.7)
TERT mutation			1.412	0.494
-	38 (29.9)	20 (25.6)
+	86 (67.7)	54 (69.2)
NA	3 (2.4)	4 (5.1)

Item	Case n	OS $ (\overline{x}\pm s)$ t/month	χ²	P value
Gender			2.406	0.121
Male	125	15.18±18.99
Female	80	15.63±13.26
Age/year			24.683	< 0.001
<55	120	18.10±20.07
≥55	85	11.48±10.04
Tumor location			1.838	0.607
Cerebellum brain stem	6	11.50±10.15
Temporo-occipital lobe	47	15.26±12.77
Frontal lobe	69	18.94±21.99
Others	83	12.71±14.04
Tumor histological type			24.437	< 0.001
Glioblastoma	93	11.74±10.90
Oligodendroglioma	33	21.61±26.19
Astrocytoma	58	17.79±18.26
Others	21	14.81±13.91
WHO grade			29.622	< 0.001
4	120	13.14±12.34
3	36	22.03±25.59
1+2	49	15.88±17.88
MGMT methylation			9.629	0.002
Negative	78	13.71±13.67
Positive	127	16.37±18.66
IDH1mut			12.563	0.002
-	113	13.54±15.80
+	76	18.64±18.72
NA	16	12.56±14.29
IDH2mut			3.226	0.199
-	180	14.70±15.14
+	6	42.17±43.90
NA	19	13.11±13.83
1p19q co-deletion			11.554	0.003
-	70	15.83±17.05
+	41	20.54±23.91
NA	94	12.74±12.23
TERT mutation			2.859	0.239
-	58	16.98±20.91
+	140	14.84±15.38
NA	7	12.14±9.34
MGMT/IDH1			18.528	0.001
Met/mut	67	18.72±19.23
Met/wt	53	14.70±18.58
Unmet/mut	9	18.11±15.20
Unmet/wt	60	12.52±12.93
NA	16	12.56±14.29
MGMT /1p19q			23.739	< 0.001
Met/co-del	36	21.81±25.08
Met/non co-del	42	16.50±18.66
Unmet/co-del	5	11.40±9.69
Unmet/non co-del	28	14.82±14.58
NA	94	12.74±12.23
MGMT/IDH1/1p19q			14.553	0.002
Met/mut/co-del	26	18.62±19.02
1 or 2 positive	64	18.77±21.79
Unmet/wt/non co-del	18	13.17±14.82
NA	97	12.64±12.12

Prognostic factor	B	SE	Wald	P value	HR	95% CI
Age	1.011	0.388	6.792	0.009	2.749	1.285-5.882
Tumor histological type	-0.057	0.224	0.064	0.801	0.945	0.609-1.467
WHO grade	-1.275	0.534	5.697	0.017	0.279	0.098-0.796
MGMT status	0.188	0.675	0.077	0.781	1.206	0.321-4.531
IDH1 status	-0.019	0.377	0.003	0.959	0.981	0.469-2.052
1p19q codel	0.378	0.308	1.500	0.221	1.459	0.797-2.67
MGMT/IDH1status	0.082	0.263	0.096	0.757	1.085	0.648-1.818
MGMT/1p19q status	0.989	0.400	6.127	0.013	2.689	1.229-5.883
MGMT/IDH1/1p19q status	-1.257	0.597	4.426	0.035	0.285	0.088-0.918