循环肿瘤细胞在原发性肝癌中的研究应用进展

doi:10.3969/j.issn.1673-8640.2019.07.020

摘要/Abstract

摘要：

原发性肝癌的致死率在恶性肿瘤中居第2位。预后不良和高复发率是原发性肝癌死亡的主要原因。循环肿瘤细胞（CTC）由实体肿瘤脱落至外周血循环中,与正常血细胞相比,其形态及标志物存在明显差异。随着富集和鉴定手段的进步,CTC对反映机体肿瘤情况、转移诊断、判断靶向疗效及指导预后具有一定意义。文章对CTC的特性、富集鉴定技术及其在原发性肝癌中的临床意义作一阐述。

关键词: 循环肿瘤细胞, 原发性肝癌, 富集鉴定, 临床研究

Abstract:

The fatality rate of primary liver cancer ranks the second in malignancy diseases,and poor prognosis and high recurrence rate are main causes for liver cancer death. Circulating tumor cells（CTC） from solid tumors fall off to peripheral blood circulation,and there are differences in morphology and markers compared with normal blood cells. With the advancement of enrichment and identification methods,CTC are of certain significance of reflecting body tumor condition,metastasis diagnosis,judging target effect and guiding prognosis. In this review,the characteristics of CTC,the technology of enrichment and identification and the clinical researches of primary liver cancer are reviewed.

Key words: Circulating tumor cell, Primary liver cancer, Enrichment and identification, Clinical research

中图分类号:

R446.1

肖潇, 高春芳. 循环肿瘤细胞在原发性肝癌中的研究应用进展[J]. 检验医学, 2019, 34(7): 661-666.

XIAO Xiao, GAO Chunfang. Research and application progress of circulating tumor cells in primary liver cancer[J]. Laboratory Medicine, 2019, 34(7): 661-666.

图/表 1

参考文献 42

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分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	文献
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	年份	作者
ISET	细胞大小	β-catenin	44	23（52.3）	巢式PCR	肿瘤分化、门脉癌栓、AFP水平、 Child分级、生存时间	2004	VONA等^[14]
无	梯度离心、流式磁珠分选	CD44⁺CD90⁺ CD45^-	36	33（91.6）	体内致瘤实验、实时荧光定量PCR	肿瘤大小	2008	YANG等^[32]
AutoMACS Pro Separator	梯度离心、磁力分选	DAPI⁺CD45^- ASGPR⁺HepPar1⁺ CK⁺AFP⁺	85	69（81.2）	免疫荧光法、 FISH、实时荧光定量PCR	肿瘤大小、门静脉血栓、Milan标准、 TNM分期	2011	XU等^[28]
无	梯度离心	CD45^- CD90⁺CD44⁺	82	56（68.3）	流式细胞术	肿瘤大小、TNM分期、复发	2011	FAN等^[15]
CellSearch 系统	阳性磁珠分选	DAPI⁺CD45^- CK⁺EpCAM⁺	87	51（58.6）	免疫荧光	AFP水平、血管侵犯、 Edmondson分级、复发	2013	SUN等^[25]
RosetteSep	阴性磁珠分选	CD45^-	30	NR	实时荧光定量 PCR
无	磁性活化细胞分选法	CD45^-EpCAM⁺	6	3（50.0）	体内致瘤实验
无	梯度离心	Lin28B⁺	96	32（33.3）	实时荧光定量 PCR	肿瘤大小、AJCC/ BCLC分期、复发	2013	CHENG等^[29]
MiniMACS Separator	梯度离心、磁力分选	DAPI⁺CD45^- HAS⁺EMT标志物	60	46（76.7）	免疫荧光法	肿瘤大小、门脉癌栓、Milan标准、 TNM分级、复发	2013	LI等^[26]
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	文献
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	年份	作者
RosetteSep	阴性磁珠分选	CD45^-EpCAM⁺	122	52（42.6）	实时荧光定量 PCR	治疗反应、复发	2014	GUO等^[40]
IsoFlux	梯度离心、磁力微流控	Hoecht 33342⁺CD45^-CK⁺	21	19（90.5）	免疫荧光法		2015	SÁNCHEZ-LORENCIO等^[41]
CellSearch 系统	阳性磁珠分选	DAPI⁺CD45^- CK⁺EpCAM⁺	20	7（35.0）	基因组测序	AFP水平、血管侵犯	2015	KELLEY等^[20]
无	阴性磁珠分选	DAPI⁺CD45^- CK⁺pERK/Akr⁺	109	101（92.7）	免疫荧光法	索拉菲尼疗效	2016	LI等^[34]
CTC-BioTChip	HA/CTS纳米膜	DAPI⁺CD45^-CK⁺	42	24（57.1）	免疫荧光法	肿瘤大小、门静脉癌栓、TNM分期	2016	WANG等^[33]
无	CTC-Chip	DAPI⁺CD45^-P- CK&CPS1⁺	36	36（100）	成球实验、免疫荧光法	索拉菲尼疗效	2016	ZHANG等^[22]
无	阳性磁珠分选	EpCAM⁺	72	43（59.7）	巢式PCR	AFP水平、分化程度	2016	JIN等^[42]
CTC-iChip	微流控芯片	CD45^-CD16^- CD66b^-	16	9（56.3）	数字PCR、 Logistic回归模型	预后分析	2017	KALINICH等^[17]
SE-iFISH	阴性磁珠分选	CK⁺EpCAM⁺CD45^-CD31^-	14	NR	免疫荧光法、 FISH、细胞大小	预后分析	2018	WANG等^[27]

分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	文献
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	年份	作者
ISET	细胞大小	β-catenin	44	23（52.3）	巢式PCR	肿瘤分化、门脉癌栓、AFP水平、 Child分级、生存时间	2004	VONA等^[14]
无	梯度离心、流式磁珠分选	CD44⁺CD90⁺ CD45^-	36	33（91.6）	体内致瘤实验、实时荧光定量PCR	肿瘤大小	2008	YANG等^[32]
AutoMACS Pro Separator	梯度离心、磁力分选	DAPI⁺CD45^- ASGPR⁺HepPar1⁺ CK⁺AFP⁺	85	69（81.2）	免疫荧光法、 FISH、实时荧光定量PCR	肿瘤大小、门静脉血栓、Milan标准、 TNM分期	2011	XU等^[28]
无	梯度离心	CD45^- CD90⁺CD44⁺	82	56（68.3）	流式细胞术	肿瘤大小、TNM分期、复发	2011	FAN等^[15]
CellSearch 系统	阳性磁珠分选	DAPI⁺CD45^- CK⁺EpCAM⁺	87	51（58.6）	免疫荧光	AFP水平、血管侵犯、 Edmondson分级、复发	2013	SUN等^[25]
RosetteSep	阴性磁珠分选	CD45^-	30	NR	实时荧光定量 PCR
无	磁性活化细胞分选法	CD45^-EpCAM⁺	6	3（50.0）	体内致瘤实验
无	梯度离心	Lin28B⁺	96	32（33.3）	实时荧光定量 PCR	肿瘤大小、AJCC/ BCLC分期、复发	2013	CHENG等^[29]
MiniMACS Separator	梯度离心、磁力分选	DAPI⁺CD45^- HAS⁺EMT标志物	60	46（76.7）	免疫荧光法	肿瘤大小、门脉癌栓、Milan标准、 TNM分级、复发	2013	LI等^[26]
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	文献
分析平台	富集方法	CTC标志物	例数	阳性例数 [例（%）]	验证方法	临床资料分析	年份	作者
RosetteSep	阴性磁珠分选	CD45^-EpCAM⁺	122	52（42.6）	实时荧光定量 PCR	治疗反应、复发	2014	GUO等^[40]
IsoFlux	梯度离心、磁力微流控	Hoecht 33342⁺CD45^-CK⁺	21	19（90.5）	免疫荧光法		2015	SÁNCHEZ-LORENCIO等^[41]
CellSearch 系统	阳性磁珠分选	DAPI⁺CD45^- CK⁺EpCAM⁺	20	7（35.0）	基因组测序	AFP水平、血管侵犯	2015	KELLEY等^[20]
无	阴性磁珠分选	DAPI⁺CD45^- CK⁺pERK/Akr⁺	109	101（92.7）	免疫荧光法	索拉菲尼疗效	2016	LI等^[34]
CTC-BioTChip	HA/CTS纳米膜	DAPI⁺CD45^-CK⁺	42	24（57.1）	免疫荧光法	肿瘤大小、门静脉癌栓、TNM分期	2016	WANG等^[33]
无	CTC-Chip	DAPI⁺CD45^-P- CK&CPS1⁺	36	36（100）	成球实验、免疫荧光法	索拉菲尼疗效	2016	ZHANG等^[22]
无	阳性磁珠分选	EpCAM⁺	72	43（59.7）	巢式PCR	AFP水平、分化程度	2016	JIN等^[42]
CTC-iChip	微流控芯片	CD45^-CD16^- CD66b^-	16	9（56.3）	数字PCR、 Logistic回归模型	预后分析	2017	KALINICH等^[17]
SE-iFISH	阴性磁珠分选	CK⁺EpCAM⁺CD45^-CD31^-	14	NR	免疫荧光法、 FISH、细胞大小	预后分析	2018	WANG等^[27]