FAM19A4、PAX1及miRNA124-2基因启动子区甲基化在宫颈病变早期诊断中的价值

doi:10.19401/j.cnki.1007-3639.2024.08.003

摘要/Abstract

摘要：

背景与目的：目前有关宫颈病变的DNA甲基化研究较多，但DNA甲基化作为宫颈病变的诊断及分流指标在临床实践中的报道较少。本研究拟探讨FAM19A4、PAX1及miRNA124-2基因启动子区甲基化在宫颈病变进展中早期诊断的价值。方法：收集2020年3月—2022年3月在郑州大学第三附属医院同时行宫颈液基细胞学（thinprep cytologic test，TCT）和HPV检测的患者的宫颈细胞学标本共129例，采用甲基化特异性PCR（methylation-specific PCR，MSP）检测不同宫颈病变中FAM19A4、PAX1及miRNA124-2基因启动子区甲基化改变情况，采用受试者工作特征（receiver operating characteristic，ROC）曲线评估3个基因甲基化改变对宫颈病变的诊断价值。本研究经郑州大学第三附属医院伦理委员会批准（伦理审批编号：2023-135-01）。 结果：根据病理学检查结果分为4组：未见上皮内病变或恶性细胞（no intraepithelial lesions or malignant lesions，NILM）组（42例）、低度鳞状上皮内病变（low grade squamous intraepithelial lesion，LSIL）组（28例）、高度鳞状上皮内病变（high grade squamous intraepithelial lesion，HSIL）组（36例）和鳞癌（squamous cervical cancer，SCC）组（23例）。随宫颈病变级别的增加，FAM19A4、PAX1及miRNA124-2基因甲基化检出率逐步增高，差异有统计学意义（P均<0.05）。在HSIL组FAM19A4、PAX1、miRNA124-2甲基化检出率分别为81.2%、80.5%和71.8%，在SCC组3种基因甲基化检出率均高达100.0%；细胞学诊断宫颈癌的ROC曲线下面积（area under curve，AUC）为0.731，诊断灵敏度和特异度分别为65.9%和80.4%； FAM19A4、PAX1及miRNA124-2基因单独诊断HSIL+（HSIL和SCC）时，PAX1甲基化诊断HSIL+的效能最高，AUC为0.925，灵敏度为92.8%，特异度为87.3%；两两联合诊断时，FAM19A4与PAX1联合诊断HSIL+的AUC为0.930，灵敏度为95.7%，特异度为87.1%；FAM19A4与miRNA124-2联合诊断HSIL+，AUC为0.895，灵敏度为97.6%，特异度为85.7%；PAX1联合miRNA124-2诊断HSIL+，AUC为0.928，灵敏度为95.7%，特异度为89.1%；PAX1、FAM19A4及miRNA124-2基因甲基化联合诊断HSIL+，AUC为0.928，灵敏度为100.0%，特异度为81.8%。结论：FAM19A4、PAX1及miRNA124-2基因启动子区甲基化诊断宫颈病变具有较高的灵敏度和特异度，有潜力成为宫颈病变早期诊断的新指标。

关键词: FAM19A4, PAX1, miRNA124-2, 甲基化, 宫颈病变

Abstract:

Background and purpose: At present, there have been many reports on the study of methylation in cervical lesions, however it is still not clinically practical as a diagnostic and shunting index of cervical lesions. In this study, we intended to investigate the early diagnostic value of FAM19A4, PAX1 and miRNA124-2 methylation in promoter region in the progression of cervical lesions. Methods: A total of 129 liquid-based cytology specimens of different grades of cervical lesions diagnosed in the Third Affiliated Hospital of Zhengzhou University from Mar. 2020 to Mar. 2022. Methylation specific PCR (MSP) was used to detect the methylation changes of FAM19A4, PAX1 and miRNA124-2 genes in different cervical lesions. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of methylation changes of the three genes in cervical lesions. This study was approved by the Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (No. 2023-135-01). Results: All the specimens were collected and divided into 4 groups according to the histological results: 42 cases of no intraepithelial lesions or malignant lesions (NILM), 28 cases of low grade squamous intraepithelial lesion (LSIL), 36 cases of high grade squamous intraepithelial lesion (HSIL), 23 cases of squamous cervical cancer (SCC). With the increase in the level of cervical lesions, FAM19A4, PAX1 and miRNA124-2 gene methylation detection rates increased gradually, and the differences were statistically significant (P<0.05). The methylation detection rates of FAM19A4, PAX1 and miRNA124-2 in HSIL group were 81.2%, 80.5% and 71.8%, respectively, and the methylation detection rates of three genes in SCC group were up to 100.0%. Cytological diagnosis of cervical cancer showed the area under curve (AUC) was 0.731, with a sensitivity and a specificity of 65.9% and 80.4%, respectively. When FAM19A4, PAX1 and miRNA124-2 were used alone to diagnose HSIL+ (HSIL and SCC), PAX1 methylation had the highest diagnostic efficiency, and the AUC was 0.925, with a sensitivity and a specificity of 92.8% and 87.3%. When the diagnosis was made in pairs, the AUC of FAM19A4 combined with PAX1 in the diagnosis of HSIL+ was 0.930, with a sensitivity and a specificity of 95.7% and 87.1%, respectively. The AUC of FAM19A4 combined with miRNA124-2 in the diagnosis of HSIL+ was 0.895, with a sensitivity and a specificity of 97.6% and 85.7%, respectively. The AUC of PAX1 combined with miRNA124-2 in the diagnosis of HSIL+ was 0.928, with a sensitivity and a specificity of 95.7% and 89.1%, respectively. When the PAX1 and FAM19A4 combined with miRNA124-2 in the diagnosis of HSIL+, the AUC was 0.928, with a sensitivity and a specificity of 100.0% and 81.8%, respectively. Conclusion: The methylation of the promoter regions of the FAM19A4, PAX1 and miRNA124-2 genes demonstrates high sensitivity and specificity in diagnosing cervical lesions, indicating the potential for becoming novel indicators for early diagnosis of cervical lesions.

Key words: FAM19A4, PAX1, miRNA124-2, Methylation, Cervical lesion

中图分类号:

R737.33

李雅, 刘慧, 任静静, 李肖甫, 智艳芳. FAM19A4、PAX1及miRNA124-2基因启动子区甲基化在宫颈病变早期诊断中的价值[J]. 中国癌症杂志, 2024, 34(8): 734-744.

LI Ya, LIU Hui, REN Jingjing, LI Xiaofu, ZHI Yanfang. Early diagnostic value of methylation in promoter region of FAM19A4, PAX1 and miRNA124-2 in cervical lesions[J]. China Oncology, 2024, 34(8): 734-744.

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Group	Pathological diagnosis				Total	x² value	P value
Group	NILM	LSIL	HSIL	SCC	Total	x² value	P value
Age/year	37.00±11.58	40.90±12.27	42.80±9.84	53.40±12.69^*	42.22±11.59	8.901	0.000
HPV
Positive	47.6% (20/42)	89.3% (25/28)	97.2% (35/36)	91.3% (21/23)	78.3% (101/129)	35.126	0.000
Negative	52.4% (22/42)	10.7% (3/28)	2.8% (1/36)	8.7% (2/23)	21.7% (28/129)
TCT
NILM	42				42
ASC-US		9	3		12
ASC-H			2	3	5
LSIL		19	2		21
HSIL			29	11	40
SCC				9	9

Gene	NILM (n=42)	LSIL (n=28)	HSIL (n=36)	SCC (n=23)	χ² value
PAX1	21.4% (9/42)	35.7% (10/28)	80.5% (29/36)	100.0% (23/23)	51.659
miRNA124-2	28.5% (12/42)	29.6% (8/27)	71.8% (23/32)	100.0% (23/23)	40.975
FAM19A4	45.2% (19/42)	51.8% (14/27)	81.2% (26/32)	100.0% (23/23)	25.688

Gene	NILM (n=42)	LSIL (n=27)	HSIL (n=32)	SCC (n=23)	χ² value
P+miR	38.1% (16/42)	44.4% (12/27)	84.3% (27/32)	100% (23/23)	34.906
P+F	47.6% (20/42)	59.3% (16/27)	87.5% (28/32)	100% (23/23)	26.104
F+miR	54.7% (23/42)	59.3% (16/27)	81.2% (26/32)	100% (23/23)	18.201
P+F+miR	57.1% (24/42)	66.7% (18/27)	87.5% (28/32)	100% (23/23)	18.476

Item	FAM19A4	PAX1	miR124-2	P+F	P+miR	miR+F	P+F+miR
Sensitivity	81.25%	80.56%	71.88%	87.50%	84.38%	81.25%	87.50%
Specificity	81.25%	78.57%	71.43%	52.38%	61.90%	45.24%	42.86%
Total coincidence rate	66.22%	79.49%	71.62%	67.57%	71.62%	60.81%	62.16%

Item	PAX1	FAM19A4	miR124-2	P+F	P+miR	miR+F	P+F+miR
Sensitivity	88.14%	89.09%	83.64%	92.73%	90.91%	89.09%	92.73%
Specificity	72.86%	52.17%	71.01%	47.83%	59.42%	43.48%	39.13%
Total coincidence rate	79.84%	68.55%	76.61%	67.74%	73.39%	63.71%	62.90%