外泌体微小RNA研究进展及诊断价值

doi:10.3969/j.issn.1673-8640.2019.12.017

摘要/Abstract

摘要：

恶性肿瘤的早期诊断对于降低恶性肿瘤患者的死亡率具有重要意义,而近年来发现外泌体微小RNA（miRNA）可以在恶性肿瘤早期诊断中发挥作用,并且其与恶性肿瘤的发生、发展关系密切,恶性肿瘤细胞所分泌的外泌体miRNA可以调节其他恶性肿瘤细胞以及肿瘤生长的环境,促进恶性肿瘤的进展。文章介绍外泌体miRNA及其提取的方法,描述其在恶性肿瘤中的机制,并分析外泌体miRNA在恶性肿瘤诊断中的优势与劣势。

关键词: 外泌体, 微小RNA, 恶性肿瘤, 诊断

Abstract:

Early diagnosis of cancer is significant for reducing the mortality of cancer patients. In recent years,exosome microRNA（miRNA） plays a role in the early diagnosis of cancer,and it is closely related to the occurrence and development of cancer. Exosome miRNA is secreted by cancer cells. It also can regulate other cancer cells and the growing environment of tumors and ultimately drive the development of cancer. This review describes exosome miRNA and its extraction methods,and further describes its mechanism in cancer,and finally analyzes the advantages and disadvantages of exosome miRNA in cancer diagnosis.

Key words: Exosome, MicroRNA, Cancer, Diagnosis

中图分类号:

R446.1

张萍, 杨波, 蔡小玲, 熊婉媛, 魏绪磐, 铁宝霞. 外泌体微小RNA研究进展及诊断价值[J]. 检验医学, 2019, 34(12): 1139-1144.

ZHANG Ping, YANG Bo, CAI Xiaoling, XIONG Wanyuan, WEI Xupan, TIE Baoxia. Research progress and diagnostic value of exosome microRNA[J]. Laboratory Medicine, 2019, 34(12): 1139-1144.

图/表 1

参考文献 42

[1]	BRAY F,FERLAY J,SOERJOMATARAM I,et al.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin,2018,68(6):394-424.
[2]	CHEN W,ZHENG R,ZHANG S,et al.Cancer incidence and mortality in China,2013[J]. Cancer Lett,2017,401:63-71.
[3]	YANG H,FU H,XU W,et al.Exosomal non-coding RNAs:a promising cancer biomarker[J]. Clin Chem Lab Med,2016,54(12):1871-1879.
[4]	WU L,QU X.Cancer biomarker detection:recent achievements and challenges[J]. Chem Soc Rev,2015,44(10):2963-2997.
[5]	THIND A,WILSON C.Exosomal miRNAs as cancer biomarkers and therapeutic targets[J]. J Extracell Vesicles,2016,5:31292.
[6]	PAN B T,JOHNSTONE R M.Fate of the transferrin receptor during maturation of sheep reticulocytes in vitro:selective externalization of the receptor[J]. Cell,1983,33(3):967-978.
[7]	SALEHI M,SHARIFI M.Exosomal miRNAs as novel cancer biomarkers:challenges and opportunities[J]. J Cell Physiol,2018,233(9):6370-6380.
[8]	VALADI H,EKSTRÖM K,BOSSIOS A,et al. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells[J]. Nat Cell Biol,2007,9(6):654-659.
[9]	WEBER J A,BAXTER D H,ZHANG S,et al.The microRNA spectrum in 12 body fluids[J]. Clin Chem,2010,56(11):1733-1741.
[10]	REKKER K,SAARE M,ROOST A M,et al.Comparison of serum exosome isolation methods for microRNA profiling[J]. Clin Biochem,2014,47(1-2):135-138.
[11]	DING M,WANG C,LU X,et al.Comparison of commercial exosome isolation kits for circulating exosomal microRNA profiling[J]. Anal Bioanal Chem,2018,410(16):3805-3814.
[12]	OSTENFELD M S,JEPPESEN D K,LAURBERG J R,et al.Cellular disposal of miR23b by RAB27-dependent exosome release is linked to acquisition of metastatic properties[J]. Cancer Res,2014,74(20):5758-5771.
[13]	YE S B,LI Z L,LUO D H,et al.Tumor-derived exosomes promote tumor progression and T-cell dysfunction through the regulation of enriched exosomal microRNAs in human nasopharyngeal carcinoma[J]. Oncotarget,2014,5(14):5439-5452.
[14]	LI J,YU J,ZHANG H,et al.Exosomes-derived MiR-302b suppresses lung cancer cell proliferation and migration via TGFβRⅡ inhibition[J]. Cell Physiol Biochem,2016,38(5):1715-1726.
[15]	LIAO J,LIU R,SHI Y J,et al.Exosome-shuttling microRNA-21 promotes cell migration and invasion-targeting PDCD4 in esophageal cancer[J]. Int J Oncol,2016,48(6):2567-2579.
[16]	SINGH R,POCHAMPALLY R,WATABE K,et al.Exosome-mediated transfer of miR-10b promotes cell invasion in breast cancer[J]. Mol Cancer,2014,13:256.
[17]	THIND A,WILSON C.Exosomal miRNAs as cancer biomarkers and therapeutic targets[J]. J Extracell Vesicles,2016,5:31292.
[18]	LIU Y,LUO F,WANG B,et al.STAT3-regulated exosomal miR-21 promotes angiogenesis and is involved in neoplastic processes of transformed human bronchial epithelial cells[J]. Cancer Lett,2016,370(1):125-135.
[19]	MAO G,LIU Y,FANG X,et al.Tumor-derived microRNA-494 promotes angiogenesis in non-small cell lung cancer[J]. Angiogenesis,2015,18(3):373-382.
[20]	ZHANG L,ZHANG S,YAO J,et al.Microenvironment-induced PTEN loss by exosomal microRNA primes brain metastasis outgrowth[J]. Nature,2015,527(7576):100-104.
[21]	GOTO T,FUJIYA M,KONISHI H,et al.An elevated expression of serum exosomal microRNA-191,-21,-451a of pancreatic neoplasm is considered to be efficient diagnostic marker[J]. BMC Cancer,2018,18(1):116.
[22]	WANG N,WANG L,YANG Y,et al.A serum exosomal microRNA panel as a potential biomarker test for gastric cancer[J]. Biochem Biophys Res Commun,2017,493(3):1322-1328.
[23]	ZHOU X,WEN W,SHAN X,et al.A six-microRNA panel in plasma was identified as a potential biomarker for lung adenocarcinoma diagnosis[J]. Oncotarget,2017,8(4):6513-6525.
[24]	LAI X,WANG M,MCELYEA S D,et al.A microRNA signature in circulating exosomes is superior to exosomal glypican-1 levels for diagnosing pancreatic cancer[J]. Cancer Lett,2017,393:86-93.
[25]	JIN X,CHEN Y,CHEN H,et al.Evaluation of tumor-derived exosomal miRNA as potential diagnostic biomarkers for early-stage non-small cell lung cancer using next-generation sequencing[J]. Clin Cancer Res,2017,23(17):5311-5319.
[26]	RODRÍGUEZ M,BAJO-SANTOS C,HESSVIK N P,et al. Identification of non-invasive miRNAs biomarkers for prostate cancer by deep sequencing analysis of urinary exosomes[J]. Mol Cancer,2017,16(1):156.
[27]	BRYZGUNOVA OE,ZARIPOV M M,SKVORTSOVA T E,et al.Comparative study of extracellular vesicles from the urine of healthy individuals and prostate cancer patients[J]. PLoS One,2016,11(6):e0157566.
[28]	MENG X,MÜLLER V,MILDE-LANGOSCH K,et al. Diagnostic and prognostic relevance of circulating exosomal miR-373,miR-200a,miR-200b and miR-200c in patients with epithelial ovarian cancer[J]. Oncotarget,2016,7(13):16923-16935.
[29]	SAMSONOV R,SHTAM T,BURDAKOV V,et al.Lectin-induced agglutination method of urinary exosomes isolation followed by mi-RNA analysis:application for prostate cancer diagnostic[J]. Prostate,2016,76(1):68-79.
[30]	ZHANG W,NI M,SU Y,et al.MicroRNAs in serum exosomes as potential biomarkers in clear-cell renal cell carcinoma[J]. Eur Urol Focus,2016,4(3):412-419.
[31]	CHIAM K,WANG T,WATSON D I,et al.Circulating serum exosomal miRNAs as potential biomarkers for esophageal adenocarcinoma[J]. J Gastrointest Surg,2015,19(7):1208-1215.
[32]	BUTZ H,NOFECH-MOZES R,DING Q,et al.Exosomal microRNAs are diagnostic biomarkers and can mediate cell-cell communication in renal cell carcinoma[J]. Eur Urol Focus,2016,2(2):210-218.
[33]	WANG J,ZHOU Y,LU J,et al.Combined detection of serum exosomal miR-21 and HOTAIR as diagnostic and prognostic biomarkers for laryngeal squamous cell carcinoma[J]. Med Oncol,2014,31(9):148.
[34]	OGATA-KAWATA H,IZUMIYA M,KURIOKA D,et al.Circulating exosomal microRNAs as biomarkers of colon cancer[J]. PLoS One,2014,9(4):e92921.
[35]	MADHAVAN B,YUE S,GALLI U,et al.Combined evaluation of a panel of protein and miRNA serum-exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity[J]. Int J Cancer,2015,136(11):2616-2627.
[36]	CAZZOLI R,BUTTITTA F,DI NICOLA M,et al.MicroRNAs derived from circulating exosomes as noninvasive biomarkers for screening and diagnosing lung cancer[J]. J Thorac Oncol,2013,8(9):1156-1162.
[37]	HIZIR M S,BALCIOGLU M,RANA M,et al.Simultaneous detection of circulating oncomiRs from body fluids for prostate cancer staging using nanographene oxide[J]. ACS Appl Mater Interfaces,2014,6(17):14772-14778.
[38]	LODES M J,CARABALLO M,SUCIU D,et al.Detection of cancer with serum miRNAs on an oligonucleotide microarray[J]. PLoS One,2009,4(7):e6229.
[39]	EICHELSER C,STÜCKRATH I,MÜLLER V,et al. Increased serum levels of circulating exosomal microRNA-373 in receptor-negative breast cancer patients[J]. Oncotarget,2014,5(20): 9650-9663.
[40]	LIU T,CHEN G,SUN D,et al.Exosomes containing miR-21 transfer the characteristic of cisplatin resistance by targeting PTEN and PDCD4 in oral squamous cell carcinoma[J]. Acta Biochimica Biophys Sin(Shanghai),2017,49(9):808-816.
[41]	QUE R,DING G,CHEN J,et al.Analysis of serum exosomal microRNAs and clinicopathologic features of patients with pancreatic adenocarcinoma[J]. World J Surg Oncol,2013,11:219.
[42]	WANG Y,ZHANG C,ZHANG P,et al.Serum exosomal microRNAs combined with alpha-fetoprotein as diagnostic markers of hepatocellular carcinoma[J]. Cancer Med,2018,7(5):1670-1679.

miRNA 种类	标本类型	分析方法	肿瘤种类	敏感性	特异性	实验组样本例数	对照组样本例数
miRNA-21^[21]	血清	qRT-PCR（TaqMan）	胰腺癌	0.81	0.82	32	22
miRNA-191^[21]				0.72	0.86
miRNA-451a^[21]				0.66	0.86
miRNA-106a-5p,miRNA-19b-3p^[22]	血清	qRT-PCR（TaqMan）	胃癌	0.95	0.90	20	20
miRNA-19b3p,miRNA-21-5p,miRNA-221-3p, miRNA-409-3p,miRNA-425-5p,miRNA-584-5p^[23]	血浆	qRT-PCR（SYBR）	肺癌	0.70	0.73	141	124
miRNA-let-7a^[24]	血浆	qRT-PCR（TaqMan）	胰腺癌	1.00	1.00	29	11
miRNA-10b^[24]				1.00	1.00
miRNA-21^[24]				1.00	1.00
miRNA-30c^[24]				1.00	1.00
miRNA-181a^[24]				1.00	1.00
miRNA-let-7b-5p,miRNA-let-7e-5p,miRNA-23a-3p, miRNA-486-5p^[25]	血浆	qRT-PCR（TaqMan）	肺癌	0.91	0.92	47	13
miRNA-196a^[26]	尿液	高通量测序	前列腺癌	0.90	1.00	20	9
miRNA-19b^[27]	尿液	qRT-PCR（TaqMan）	前列腺癌	0.94	0.80	35	35
miRNA-25^[27]				0.71	0.69
miRNA-125b^[27]				0.86	0.66
miRNA-205^[27]				0.57	0.74
miRNA-200a^[28]	血清	qRT-PCR（TaqMan）	卵巢癌	0.84	0.90	163	20
miRNA-200b^[28]				0.53	1.00
miRNA-200c^[28]				0.31	1.00
miRNA-200a,miRNA-200b,miR200c^[28]				0.88	0.90
miRNA-21-5p^[29]	尿液	qRT-PCR（SYBR）	前列腺癌	0.66	0.94	35	35
miRNA-141-5p^[29]				0.66	0.94
miRNA-574-3p^[29]				0.86	1.00
miRNA-210^[30]	血清	qRT-PCR（SYBR）	肾细胞癌	0.70	0.63	82	80
miRNA-1233^[30]				0.82	0.76
miRNA-16-5p^[31]	血清	miRNA芯片	食管癌	0.78	0.79	18	29
miRNA-16-5p,miRNA-25-3p/miRNA-320a, let-7e-5p/miR15b-5p, miRNA-30a-5p/miRNA-324-5p, miRNA-17-5p/miRNA-194-5p^[31]				0.94	1.00
miRNA-126-3p,miRNA-449a^[32]	尿液	qRT-PCR（TaqMan）	肾细胞癌	0.79	0.71	109	51
miRNA-126-3p,miRNA-34b-5p^[32]				0.84	0.67
miRNA-21^[33]	血清	qRT-PCR（SYBR）	喉癌	0.69	0.77	52	49
miRNA-23a^[34]	血清	miRNA芯片	结肠癌	0.92	1.00	88	11
miRNA-1246^[34]				0.95	0.91
miRNA-1246,miRNA-4644,miRNA-3976,miRNA-4306^[35]	血清	qRT-PCR（SYBR）	胰腺癌	0.80	1.00	12	75
miRNA-151a-5p,miRNA-30a-3p,miRNA-200b-5p, miRNA-629,miRNA-100,miRNA-154-3p^[36]	血浆	qRT-PCR（SYBR）	肺癌	0.96	0.60	50	30

miRNA 种类	标本类型	分析方法	肿瘤种类	敏感性	特异性	实验组样本例数	对照组样本例数
miRNA-21^[21]	血清	qRT-PCR（TaqMan）	胰腺癌	0.81	0.82	32	22
miRNA-191^[21]				0.72	0.86
miRNA-451a^[21]				0.66	0.86
miRNA-106a-5p,miRNA-19b-3p^[22]	血清	qRT-PCR（TaqMan）	胃癌	0.95	0.90	20	20
miRNA-19b3p,miRNA-21-5p,miRNA-221-3p, miRNA-409-3p,miRNA-425-5p,miRNA-584-5p^[23]	血浆	qRT-PCR（SYBR）	肺癌	0.70	0.73	141	124
miRNA-let-7a^[24]	血浆	qRT-PCR（TaqMan）	胰腺癌	1.00	1.00	29	11
miRNA-10b^[24]				1.00	1.00
miRNA-21^[24]				1.00	1.00
miRNA-30c^[24]				1.00	1.00
miRNA-181a^[24]				1.00	1.00
miRNA-let-7b-5p,miRNA-let-7e-5p,miRNA-23a-3p, miRNA-486-5p^[25]	血浆	qRT-PCR（TaqMan）	肺癌	0.91	0.92	47	13
miRNA-196a^[26]	尿液	高通量测序	前列腺癌	0.90	1.00	20	9
miRNA-19b^[27]	尿液	qRT-PCR（TaqMan）	前列腺癌	0.94	0.80	35	35
miRNA-25^[27]				0.71	0.69
miRNA-125b^[27]				0.86	0.66
miRNA-205^[27]				0.57	0.74
miRNA-200a^[28]	血清	qRT-PCR（TaqMan）	卵巢癌	0.84	0.90	163	20
miRNA-200b^[28]				0.53	1.00
miRNA-200c^[28]				0.31	1.00
miRNA-200a,miRNA-200b,miR200c^[28]				0.88	0.90
miRNA-21-5p^[29]	尿液	qRT-PCR（SYBR）	前列腺癌	0.66	0.94	35	35
miRNA-141-5p^[29]				0.66	0.94
miRNA-574-3p^[29]				0.86	1.00
miRNA-210^[30]	血清	qRT-PCR（SYBR）	肾细胞癌	0.70	0.63	82	80
miRNA-1233^[30]				0.82	0.76
miRNA-16-5p^[31]	血清	miRNA芯片	食管癌	0.78	0.79	18	29
miRNA-16-5p,miRNA-25-3p/miRNA-320a, let-7e-5p/miR15b-5p, miRNA-30a-5p/miRNA-324-5p, miRNA-17-5p/miRNA-194-5p^[31]				0.94	1.00
miRNA-126-3p,miRNA-449a^[32]	尿液	qRT-PCR（TaqMan）	肾细胞癌	0.79	0.71	109	51
miRNA-126-3p,miRNA-34b-5p^[32]				0.84	0.67
miRNA-21^[33]	血清	qRT-PCR（SYBR）	喉癌	0.69	0.77	52	49
miRNA-23a^[34]	血清	miRNA芯片	结肠癌	0.92	1.00	88	11
miRNA-1246^[34]				0.95	0.91
miRNA-1246,miRNA-4644,miRNA-3976,miRNA-4306^[35]	血清	qRT-PCR（SYBR）	胰腺癌	0.80	1.00	12	75
miRNA-151a-5p,miRNA-30a-3p,miRNA-200b-5p, miRNA-629,miRNA-100,miRNA-154-3p^[36]	血浆	qRT-PCR（SYBR）	肺癌	0.96	0.60	50	30