miR-2355-3p、miR-337-3p和miR-99a-5p检测在头颈部鳞状细胞癌早期筛查中的价值

doi:10.16150/j.1671-2870.2025.02.012

摘要/Abstract

摘要：

目的： 探讨miR-2355-3p、miR-337-3p和miR-99a-5p在头颈部鳞状细胞癌（head and neck squamous cell carcinoma, HNSCC）早期筛查中的价值。 方法： 收集2023年7月至2024年7月在上海交通大学医学院附属第九人民医院首次就诊的60例HNSCC患者（HNSCC组）及60例健康体检人群（正常对照组）的血清样本及临床病理资料，以平行检测方式检测miR-2355-3p、miR-337-3p和miR-99a-5p的相对表达量。采用受试者操作特征（receiver opera-ting characteristic, ROC）曲线评估3种miRNA对HNSCC的筛查、诊断效能。 结果： 相较于正常对照组，HNSCC组患者血清中miR-2355-3p和miR-337-3p的表达显著上调（P<0.001），而miR-99a-5p的表达显著下调（P=0.002）。肿瘤>2 cm与≤2 cm、肿瘤分期Ⅰ-Ⅱ期与Ⅲ-Ⅳ期、有及无颈部淋巴结转移的HNSCC患者间，血清中miR-2355-3p及miR-337-3p的表达量差异有统计学意义（P<0.05），不同年龄和性别的HNSCC患者血清中3种miRNA的表达量差异均无统计学意义（P>0.05）。miR-2355-3p、miR-337-3p及miR-99a-5p单项检测诊断HNSCC的最佳截断值分别为0.0867、0.1031和0.1251，曲线下面积（area under the curve, AUC）分别为0.892、0.877和0.686。3项miRNA联合检测的AUC（0.954）在高于各单项检测的AUC（P<0.05）的同时，还高于miR-2355-3p、miR-337-3p 2项联合检测的AUC（0.898）（P<0.05）。3项miRNA联合检测早期筛查Ⅰ-Ⅱ期HNSCC的AUC为0.923。 结论： miR-2355-3p和miR-337-3p与HNSCC肿瘤的分期、大小及淋巴结转移相关，miR-2355-3p、miR-337-3p及miR-99a-5p联合检测可能用于HNSCC早期筛查。

关键词: miR-2355-3p, miR-337-3p, miR-99a-5p, 生物标志物, 头颈部鳞状细胞癌

Abstract:

Objective To explore the value of miR-2355-3p, miR-337-3p, and miR-99a-5p in the early screening of head and neck squamous cell carcinoma (HNSCC). Methods Serum samples and clinicopathological data were collected from 60 newly diagnosed HNSCC patients (HNSCC group) and 60 healthy individuals (normal control group) who visited the Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine between July 2023 and July 2024. The relative expression levels of miR-2355-3p, miR-337-3p, and miR-99a-5p were determined using parallel detection. The HNSCC screening and diagnostic efficacy of these three miRNAs was evaluated using receiver operating characteristic (ROC) curves. Results Compared with the normal control group, serum levels of miR-2355-3p and miR-337-3p were significantly upregulated in the HNSCC group (P<0.001), while miR-99a-5p was significantly downregulated (P=0.002). The expression levels of miR-2355-3p and miR-337-3p differed significantly among HNSCC patients with tumor size >2 cm versus ≤2 cm, clinical stage Ⅰ–Ⅱ versus stage Ⅲ–Ⅳ, and with versus without cervical lymph node metastasis (P<0.05). However, no significant differences in miRNA expression were found across different age or gender groups (P>0.05). The optimal cutoff values for diagnosing HNSCC were 0.0867 for miR-2355-3p, 0.1031 for miR-337-3p, and 0.1251 for miR-99a-5p, with corresponding areas under the curve (AUCs) of 0.892, 0.877, and 0.686, respectively. The combined detection of the three miRNAs yielded an AUC of 0.954, significantly higher than those of individual markers (P<0.05) and the combination of miR-2355-3p and miR-337-3p (AUC=0.898, P<0.05). The AUC for the combined detection in early-stage (stage Ⅰ–Ⅱ) HNSCC was 0.923. Conclusions miR-2355-3p and miR-337-3p are associated with the tumor stage, size, and lymph node metastasis of HNSCC. The combined detection of miR-2355-3p, miR-337-3p, and miR-99a-5p may serve as a potential method for early screening of HNSCC.

Key words: miR-2355-3p, miR-337-3p, miR-99a-5p, Biomarkers, Head and neck squamous cell carcinoma

中图分类号:

R730.4

刘敬浩, 郭海艳, 甘桂芳, 陈福祥. miR-2355-3p、miR-337-3p和miR-99a-5p检测在头颈部鳞状细胞癌早期筛查中的价值[J]. 诊断学理论与实践, 2025, 24(02): 204-211.

LIU Jinghao, GUO Haiyan, GAN Guifang, CHEN Fuxiang. Value of miR-2355-3p，miR-337-3p and miR-99a-5p detection in early screening of head and neck squamous cell carcinoma[J]. Journal of Diagnostics Concepts & Practice, 2025, 24(02): 204-211.

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Primer	Primer sequence（5’→3’）
miR-2355-5p RT	5’-GTCGTATCCAGTGCAGGGTCCGAGGTA TTCGCACTGGATACGACATCTCC-3’
miR-2355-5p Forward	5’-AAGCGCCTATTGTCCTTGCTGT-3’
miR-2355-5p Reverse	5’-ATCCAGTGCAGGGTCCGAGG-3’
miR-337-3p RT	5’-GTCGTATCCAGTGCAGGGTCCGAGGT ATTCGCACTGGATACGACGAAGAA-3’
miR-337-3p Forward	5’-AAGCGACCCTCCTATATGATGC-3’
miR-337-3p Reverse	5’-GTCGTATCCAGTGCAGGGT-3’
miR-99a-5p RT	5’-GTCGTATCCAGTGCAGGGTCCGAGGT ATTCGCACTGGATACGACCACAAG-3’
miR-99a-5p Forward	5’-AACACGTGAACCCGTAGATCCG-3’
miR-99a-5p Reverse	5’-ATCCAGTGCAGGGTCCGAGG-3’
U6 Forward	5’-CTACTCTTTCTTCAAATCCC-3’
U6 Reverse	5’-GCTCTACCACCACATCCT-3’

Groups	N	miR-2355-3p	miR-337-3p	miR-99a-5p
NC	60	0.062±0.051	0.070±0.044	0.311±0.202
HNSCC	60	0.222±0.142	0.190±0.103	0.207±0.152
NC vs HNSCC		P<0.001（t=-8.211）	P<0.001（t=-8.264）	P=0.002（t=3.190）

Indice	N	miR-2355-3p		miR-337-3p		miR-99a-5p
Indice	N	$\bar{x}\pm s$	P value（t）	$\bar{x}\pm s$	Pvalue（t）	$\bar{x}\pm s$	P value（t）
Sex			0.858（-0.180）		0.861（0.176）		0.471（0.726）
Male	33	0.219±0.144		0.192±0.113		0.220±0.146
Female	27	0.226±0.142		0.187±0.092		0.191±0.160
Age			0.947（-0.066）		0.710（-0.373）		0.211（-1.266）
≥60	35	0.221±0.133		0.185±0.098		0.186±0.138
<60	25	0.223±0.156		0.196±0.112		0.236±0.168
TNM Stage
Stage Ⅰ-Ⅱ	36	0.128±0.055	<0.001（-9.704）	0.136±0.067	<0.001（-5.997）	0.219±0.165	0.446（0.767）
Stage Ⅲ-Ⅳ	24	0.363±0.110		0.271±0.096		0.188±0.131
Lymph nodes metastasis			<0.001（9.773）		<0.001（5.782）		0.597（-0.532）
With	23	0.369±0.108		0.272±0.098		0.193±0.131
Without	37	0.130±0.057		0.138±0.068		0.215±0.164
Tumor size			<0.001（4.484）		<0.001（5.331）		0.595（0.535）
＞2 cm	35	0.282±0.123		0.239±0.082		0.216±0.146
≤2 cm	25	0.138±0.123		0.120±0.090		0.194±0.161

Indice	Combined detection equation	AUC(95%CI）	Sensitivity （%）	Specificity （%）	Youden index
miR-2355-3p		0.892(0.822-0.941)	86.67	80.00	0.6667
miR-337-3p		0.877(0.805-0.930)	81.67	81.67	0.6333
miR-99a-5p		0.686(0.595-0.768)	38.33	93.33	0.3167
miR-2355-3p + miR-337-3p	Logit(P)=-2.925 + 13.398miR-2355-3p + 12.424miR-337-3p	0.898(0.830-0.946)	85.00	80.00	0.6500
miR-337-3p + miR-99a-5p	Logit(P)=-1.680 + 36.809miR-337-3p - 9.040miR-99a-5p	0.927(0.865-0.967)	86.67	91.67	0.7833
miR-2355-3p + miR-99a-5p	Logit(P)=-1.456 + 43.541miR-2355-3p - 11.258miR-99a-5p	0.952(0.897-0.983)	90.00	95.00	0.8500
miR-2355-3p + miR-99a-5p + miR-337-3p	Logit(P)=-1.796 + 33.979miR-2355-3p + 12.457miR-337-3p - 11.262*miR-99a-5p	0.954(0.899-0.984)	91.67	96.67	0.8833

Indice	Combined detection equation	AUC(95%CI）	Sensitivity （%）	Specificity （%）	Youden index
miR-2355-3p		0.820(0.728-0.891)	80.56	78.33	0.5889
miR-337-3p		0.806(0.712-0.879)	69.44	81.67	0.5111
miR-99a-5p		0.658(0.554-0.752)	38.89	93.33	0.3222
miR-2355-3p + miR-337-3p	Logit(P)=-2.925 + 13.398miR-2355-3p + 12.424miR-337-3p	0.832(0.742-0.901)	86.11	70.00	0.5611
miR-337-3p + miR-99a-5p	Logit(P)=-1.680 + 36.809miR-337-3p - 9.040miR-99a-5p	0.880(0.798-0.937)	77.78	91.67	0.6944
miR-2355-3p + miR-99a-5p	Logit(P)=-1.456 + 43.541miR-2355-3p - 11.258miR-99a-5p	0.920(0.846-0.965)	83.33	95.00	0.7833
miR-2355-3p + miR-99a-5p + miR-337-3p	Logit(P)=-1.796 + 33.979miR-2355-3p + 12.457miR-337-3p - 11.262*miR-99a-5p	0.923(0.850-0.967)	86.11	96.67	0.8278