长链非编码RNA ENST00000489676在超声评估甲状腺乳头状癌颈部淋巴结转移中的应用

doi:10.16150/j.1671-2870.2022.04.016

摘要/Abstract

摘要：

目的：分析超声引导下细针穿刺活检（ultrasound fine needle aspiration biopsy, US-FNAB）组织中长链非编码RNA（long non-coding RNA,lncRNA）ENST0000048967在超声诊断甲状腺乳头状癌（papillary thyroid carcinoma,PTC）淋巴结转移中的应用。方法：选取2020年6月至2020年12月于我院行US-FNAB且经手术病理证实为PTC颈部淋巴结转移的患者110例（转移组）,另设110例无颈部淋巴结转移的PTC患者作为对照（非转移组）。采用聚合酶链反应（polymerase chain reaction,PCR）检测PTC穿刺标本中ENST00000489676的相对表达水平,比较转移组与非转移组（各110例）表达水平差异,分析ENST00000489676表达水平与颈部淋巴结转移间的相关性。同时利用受试者操作特征（receiver operator characteristic,ROC）曲线计算穿刺标本ENST00000489676水平诊断PTC转移的最佳临界值（cutoff值）,计算ENST00000489676检测与超声联合诊断PTC淋巴结转移的灵敏度、特异度及准确率。结果：转移组ENST00000489676相对表达水平显著低于非转移组（2.61±1.59,3.69±2.18）（t=-4.198,P<0.001）,统计结果显示ENST00000489676低表达与PTC淋巴结转移显著相关。ROC曲线显示,ENST00000489676在穿刺标本中相对表达水平的最佳临界值为2.0950,以ENST00000489676低于该值诊断PTC淋巴结转移的灵敏度及准确率均高于超声单独诊断（灵敏度61.11%比17.27%;特异度63.39%比86.36%;准确率62.27%比51.82%）。结论：低表达ENST00000489676与PTC淋巴结转移显著相关,ENST00000489676作为分子标志物可应用于辅助超声诊断。

关键词: 甲状腺乳头状癌, 淋巴结转移, ENST00000489676, 超声, 超声引导下细针穿刺活检

Abstract:

Objective: To analyze the value of lncRNA ENST0000048967 detection in biopsy tissue of papillary thyroid carcinoma (PTC) by ultrasound-guided fine-needle aspiration (US-FNA) for diagnosing cervical lymph node metastasis. Methods: A total of 110 PTC patients with pathologically confirmed cervical lymph node metastasis in Ruijin hospital during June 2020 to December 2020, were enrolled (study group). Another 110 PTC patients without cervical lymph node metastasis were served as the control group. The expression level of ENST00000489676 in FNA biopsy samples from 220 patients were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The correlation of ENST00000489676 expression with lymph node metastasis in PTC patients was explored. ROC curve was used to calculate the cutoff value of ENST00000489676 expression level for diagnosing cervical lymph node metastasis of PTC in puncture specimens. The sensitivity, specificity and accuracy of ultrasonography alone and combined with ENST00000489676 detection in FNA biopsy samples for diagnosis of lymph node metastasis were also calculated respectively. Results: The expression of ENST0000048967 in study group was significantly lower than that in non-lymph node metastasis group(t=-4.198, P<0.001). The statistical results showed that the low expression of ENST000000489676 was positively correlated with PTC lymph node metastasis. The optimal cutoff value of ENST00000489676 for diagnosing cervical lymph node metastasis of PTC in the FNAB specimen was 2.0950 by ROC curve. The sensitivity and accuracy of ultrasonography combined with ENST00000489676 detection for diagnosising lymph node metastasis were higher than those of ultrasonography alone, respectively(sensitivity, 61.11% and 17.27%; specificity, 63.39% and 86.36%; accuracy, 62.27%, and 51.82%). Conclusions: Low expression of ENST00000489676 is significantly correlated with lymph node metastasis of PTC. ENST00000489676 may serve as a molecular marker to assist ultrasonography diagnosis of cervical lymph node metastasis for PTC.

Key words: Papillary thyroid carcinoma, Lymph node metastasis, ENST00000489676, Ultrasonography, US-fine needle aspiration biopsy

中图分类号:

R541.7

王文涵, 夏蜀珺, 詹维伟. 长链非编码RNA ENST00000489676在超声评估甲状腺乳头状癌颈部淋巴结转移中的应用[J]. 诊断学理论与实践, 2022, 21(04): 514-519.

WANG Wenhan, XIA Shujun, ZHAN Weiwei. Application of long non-coding RNA ENST00000489676 detection in ultrasonographic evaluation of cervical lymph node metastasis in papillary thyroid carcinoma[J]. Journal of Diagnostics Concepts & Practice, 2022, 21(04): 514-519.

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参考文献 25

[1]	Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries[J]. CA Cancer J Clin, 2021, 71(3):209-249. doi: 10.3322/caac.21660 URL
[2]	Haugen BR, Alexander EK, Bible KC, et al. 2015 Ame-rican Thyroid Association Management Guidelines for Adult Patients with Thyroid Nodules and Differentiated Thyroid Cancer: The American Thyroid Association Guidelines Task Force on Thyroid Nodules and Differentiated Thyroid Cancer[J]. Thyroid, 2016, 26(1):1-133. doi: 10.1089/thy.2015.0020 URL
[3]	Limberg J, Ullmann TM, Stefanova D, et al. Does Aggressive Variant Histology Without Invasive Features Predict Overall Survival in Papillary Thyroid Cancer?: A National Cancer Database Analysis[J]. Ann Surg, 2021, 274(3):e276-e281. doi: 10.1097/SLA.0000000000003632 URL
[4]	O′Grady TJ, Gates MA, Boscoe FP. Thyroid cancer incidence attributable to overdiagnosis in the United States 1981-2011[J]. Int J Cancer, 2015, 137(11):2664-2673. doi: 10.1002/ijc.29634 pmid: 26069163
[5]	Colonna M, Uhry Z, Guizard AV, et al. Recent trends in incidence, geographical distribution, and survival of pa-pillary thyroid cancer in France[J]. Cancer Epidemiol, 2015, 39(4):511-518. doi: 10.1016/j.canep.2015.04.015 pmid: 26003877
[6]	Feng JW, Yang XH, Wu BQ, et al. Predictive factors for central lymph node and lateral cervical lymph node metastases in papillary thyroid carcinoma[J]. Clin Transl Oncol, 2019, 21(11):1482-1491. doi: 10.1007/s12094-019-02076-0 pmid: 30879178
[7]	Qu N, Zhang L, Ji QH, et al. Risk Factors for Central Compartment Lymph Node Metastasis in Papillary Thyroid Microcarcinoma: A Meta-Analysis[J]. World J Surg, 2015, 39(10):2459-2470. doi: 10.1007/s00268-015-3108-3 pmid: 26099728
[8]	Mulla M, Schulte KM. Central cervical lymph node metastases in papillary thyroid cancer: a systematic review of imaging-guided and prophylactic removal of the central compartment[J]. Clin Endocrinol (Oxf), 2012, 76(1):131-136. doi: 10.1111/j.1365-2265.2011.04162.x URL
[9]	Lee EW, Chen C, Sauk S, et al. How diagnostic is ultrasound-guided neck mass biopsy (fine-needle capillary sampling biopsy technique)?: evaluation of 132 nonthyroid neck mass biopsies with pathologic analysis over 7 years at a single institution[J]. J Ultrasound Med, 2009, 28(12):1679-1684. pmid: 19933482
[10]	Kopp F, Mendell JT. Functional Classification and Experimental Dissection of Long Noncoding RNAs[J]. Cell, 2018, 172(3):393-407. doi: S0092-8674(18)30048-5 pmid: 29373828
[11]	Boon RA, Jaé N, Holdt L, et al. Long Noncoding RNAs: From Clinical Genetics to Therapeutic Targets?[J]. J Am Coll Cardiol, 2016, 67(10):1214-1226. doi: S0735-1097(16)00224-2 pmid: 26965544
[12]	Winkler L, Dimitrova N. A mechanistic view of long noncoding RNAs in cancer[J]. Wiley Interdiscip Rev RNA, 2022, 13(3):e1699.
[13]	Gutschner T, Diederichs S. The hallmarks of cancer: a long non-coding RNA point of view[J]. RNA Biol, 2012, 9(6):703-719. doi: 10.4161/rna.20481 pmid: 22664915
[14]	Yan H, Bu P. Non-coding RNA in cancer[J]. Essays Biochem, 2021, 65(4):625-639. doi: 10.1042/EBC20200032 pmid: 33860799
[15]	陈滢, 何莉莉, 蒋锋. 长链非编码RNA FENDRR在结肠直肠癌中的表达及意义[J]. 诊断学理论与实践, 2018, 17(1):87-91.
	He LL, Jiang F. Expression of long non-coding RNA FENDRR in human colorectal cancer and its correlation with prognosis[J]. J Diagn Concepts & Pract, 2018, 17(1):87-91.
[16]	Zhou T, Zhong M, Zhang S, et al. LncRNA CASC2 expression is down- regulated in papillary thyroid cancer and promotes cell invasion by affecting EMT pathway[J]. Cancer Biomark, 2018, 23(2):185-191. doi: 10.3233/CBM-181198 pmid: 30175973
[17]	Xia E, Bhandari A, Shen Y, et al. LncRNA CCND2-AS1 promotes proliferation, migration, and invasion in papillary thyroid carcinoma[J]. Biochem Biophys Res Commun, 2018, 496(2):628-632. doi: 10.1016/j.bbrc.2018.01.102 URL
[18]	Xiang J, Guan Y, Bhandari A, et al. LINC00511 influe-nces cellular proliferation through cyclin-dependent kinases in papillary thyroid carcinoma[J]. J Cancer, 2020, 11(2):450-459. doi: 10.7150/jca.35364 URL
[19]	Wang W, Xia S, Zhan W. The Long Non-Coding RNA ENST00000489676 Influences Papillary Thyroid Cancer Cell Proliferation and Invasion through Regulating MiR-922[J]. J Cancer, 2019, 10(22):5434-5446. doi: 10.7150/jca.31751 pmid: 31632488
[20]	Gupta RA, Shah N, Wang KC, et al. Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis[J]. Nature, 2010, 464(7291):1071-1076. doi: 10.1038/nature08975 URL
[21]	Yang Z, Zhou L, Wu LM, et al. Overexpression of long non-coding RNA HOTAIR predicts tumor recurrence in hepatocellular carcinoma patients following liver transplantation[J]. Ann Surg Oncol, 2011, 18(5):1243-1250. doi: 10.1245/s10434-011-1581-y pmid: 21327457
[22]	Liu FT, Qiu C, Luo HL, et al. The association of HOTAIR expression with clinicopathological features and prognosis in gastric cancer patients[J]. Panminerva Med, 2016, 58(2):167-174. pmid: 26964077
[23]	Wang Y, Xue D, Li Y, et al. The Long Noncoding RNA MALAT-1 is A Novel Biomarker in Various Cancers: A Meta-analysis Based on the GEO Database and Literature[J]. J Cancer, 2016, 7(8):991-1001. doi: 10.7150/jca.14663 pmid: 27313790
[24]	Zhang R, Xia Y, Wang Z, et al. Serum long non coding RNA MALAT-1 protected by exosomes is up-regulated and promotes cell proliferation and migration in non-small cell lung cancer[J]. Biochem Biophys Res Commun, 2017, 490(2):406-414. doi: 10.1016/j.bbrc.2017.06.055 URL
[25]	Goyal B, Yadav SRM, Awasthee N, et al. Diagnostic, prognostic, and therapeutic significance of long non-co-ding RNA MALAT1 in cancer[J]. Biochim Biophys Acta Rev Cancer, 2021, 1875(2):188502. doi: 10.1016/j.bbcan.2021.188502 URL

Gene/shRNA		序列（5′→3′）
ENST00000489676	正向引物	GGCAGCTGACAGGGCTTT
	逆向引物	GCAGTCCACCCTGTCCTTG
GAPDH	正向引物	AAGGTGAAGGTCGGAGTCAAC
	逆向引物	GGGGTCATTGATGGCAACAATA

特征	高表达组（113例）	低表达组（107例）	P值	χ²值
性别			0.214	1.543
男	29(13.18%)	20(9.09%)
女	84(38.18%)	87(39.55%)
年龄（岁）			0.962	0.002
<45	69(31.36%)	65(29.55%)
≥45	44(20.00%)	42(19.09%)
最大径（mm）	11.31±5.88	9.88±6.65
<10	54(24.55%)	64(29.09%)	0.074	3.196
≥10	59(26.82%)	43(19.54%)
多灶癌			0.555	0.349
否	73(33.18%)	65(29.55%)
是	40(18.18%)	42(19.09%)
形态			0.883	0.022
不规则	91(41.36%)	87(39.55%)
规则	22(10.00%)	20(9.09%)
A/T			0.198	1.658
<1	76(34.55%)	63(28.63%)
≥1	37(16.82%)	44(20.00%)
边缘			0.685	0.165
不光整	104(47.27%)	100(45.45%)
光整	9(4.09%)	7(3.19%)
内部回声			1.000	0.000
非低回声	2(0.91%)	1(0.46%)
低回声	111(50.45%)	106(48.18%)
内部结构			0.962	0.002
非实性	1(0.46%)	2(0.91%)
实性	112(50.91%)	105(47.72%)
钙化			0.376	0.784
非微钙化	43(19.55%)	47(21.36%)
微钙化	70(31.82%)	60(27.27%)
血供			0.241	4.195
无	8(3.64%)	13(5.91%)
低	78(35.45%)	60(27.27%)
中	21(9.55%)	26(11.82%)
高	6(2.72%)	8(3.64%)
转移			0.001	13.215
有	42(19.09%)	66(30.00%)
无	71(32.27%)	41(18.64%)