前列腺癌患者外周血循环肿瘤细胞分型及自噬蛋白的表达对于远处转移的诊断价值

doi:10.3877/cma.j.issn.1674-3253.2021.05.003

目的

远处转移为前列腺癌的主要死亡原因，如何识别高转移风险的前列腺癌具有重要临床意义。本研究旨在通过CanPatrol技术检测前列腺癌外周血循环细胞（CTCs），并探究CTCs上皮间质转化（EMT）情况以及自噬相关蛋白MAP1S表达情况与前列腺癌远处转移等临床病理特征的关系。

方法

纳入2017年11月至2018年11月中山大学孙逸仙纪念医院泌尿外科确诊且未经治疗的前列腺癌患者。用CanPatrol系统检测外周血循环肿瘤细胞并根据上皮及间质标记物表达情况以及MAP1S表达情况进行分型，分析CTCs分型与前列腺癌临床病理特征的关系。

结果

根据纳入及排除标准，共纳入前列腺癌病例26例，其中CTCs检出率100%，间质型CTCs检出率92.30%。MAP1S阳性率88.46%，MAP1S阳性CTCs个数3.50（5.25）。分析CTCs分型及临床病理特征的关系发现，MAP1S阳性率高者远处转移风险高，间质型CTCs比例高者远处转移风险高，Gleason评分更高。

结论

本研究采用一种较新的CTCs检测系统，将间质标志物纳入检测体系，并同时检测了自噬标志物MAP1S表达情况。发现高间质型CTCs比例与远处转移及高Gleason评分相关，高MAP1S表达率与远处转移相关。提示检测外周血循环肿瘤细胞的EMT标志物与自噬标志物或可协助前列腺癌转移的早期诊断。

关键词: 前列腺癌, 循环肿瘤细胞, 远处转移, 上皮间质转化, 自噬

Objective

Distant metastasis is a main cause of death of prostate cancer. How to identify prostate cancer with a high risk of metastasis is of great important in clinical practice. To clarify the significance of circulating tumor cells (CTCs) undergoing epithelial mesenchymal transition (EMT) and CTCs expressing MAP1S in patients with prostate cancer, we used an advanced CanPatrol CTC-enrichment technique and in situ hybridization to enrich and classify CTC from blood samples.

Methods

Patients with prostate cancer who were diagnosed by biopsy or postoperation pathology without any anti-tumor treatment were enrolled in study during November 2017 to November 2018. Enrichment of peripheral blood tumor cells was performed using CanPatrolTM CTCs enrichment technique and fluorescence in situ hybridization was performed to detect epithelial and mesenchymal marker and MAP1S expression. The captured circulating tumor cells was divided into epithelial type (expressing epithelial markers only), hybrid type (simultaneous expression of epithelial and mesenchymal markers), and mesenchymal type (expressing mesenchymal markers only) to investigate their relevance to the clinical features of prostate cancer.

Result

A total of 26 patients were included, all patients was CTC positive, 92.30% patients was hybrid CTC positive. Average MAP1S positive CTCs count was 3.50(5.25). Increased MAP1S positive rate was associated with increased distant metastasis risk, and increased hybrid CTCs proportion associated with increased distant metastasis risk and increased Gleason grade.

Conclusion

The EMT process and MAP1s expression rate of CTCs is related to distant metastases of prostate cancer. CTC typing is expecting to employ as an early diagnostic tool for non-invasive diagnosis of prostate cancer metastases.

Key words: Prostate cance, Circulating tumor cells, Distant metastasis, EMT, MAP1S

图1 外周血循环肿瘤细胞分型标准注：a为上皮间质转化分类；b为MAP1S表达情况分类

表1 外周血CTCs分型标准

	CD45(白色)	上皮型探针(红色)	间质型探针(绿色)
	CD45(白色)	EpCAM、CK8/18/19	vimentin、twist
白细胞	+	-	-
上皮型CTCs	-	+	-
间质型CTCs	-	+/-	+

表1 外周血CTCs分型标准

	CD45(白色)	上皮型探针(红色)	间质型探针(绿色)
	CD45(白色)	EpCAM、CK8/18/19	vimentin、twist
白细胞	+	-	-
上皮型CTCs	-	+	-
间质型CTCs	-	+/-	+

图2 ROC曲线评估间质型CTCs百分比对转移情况的诊断价值

表2 前列腺癌外周血CTCs分型与临床病理特征的关系(例)

		总CTCs计数		P值	MAP1S⁺CTCs百分比		P值	间质型CTCs百分比		P值
		≤5	>5	P值	≤0.6%	>0.6%	P值	≤0.8%	>0.8%	P值
原发肿瘤情况	T1、T2	4	9	0.567	7	6	0.543	7	6	0.302^*
原发肿瘤情况	T3、T4	6	6	0.567	5	7	0.543	4	8	0.302^*
淋巴结情况	N0	7	14 0	0.555	10 0	11 0	0.619	11 0	10 0	0.42^*
淋巴结情况	N1	3	2	0.555	3	2	0.619	1	4	0.42^*
转移情况	M0	4	11 0	0.228	10 0	5	0.047*	10 0	5	0.021^*
转移情况	M1	6	5	0.228	3	8	0.047*	2	9	0.021^*
Gleason评分	3+4=7	2	4	0.702	1	5	0.129	5	1	0.047^*
Gleason评分	4+3=7	8	110	0.702	120	7	0.129	7	120	0.047^*

表2 前列腺癌外周血CTCs分型与临床病理特征的关系(例)

		总CTCs计数		P值	MAP1S⁺CTCs百分比		P值	间质型CTCs百分比		P值
		≤5	>5	P值	≤0.6%	>0.6%	P值	≤0.8%	>0.8%	P值
原发肿瘤情况	T1、T2	4	9	0.567	7	6	0.543	7	6	0.302^*
原发肿瘤情况	T3、T4	6	6	0.567	5	7	0.543	4	8	0.302^*
淋巴结情况	N0	7	14 0	0.555	10 0	11 0	0.619	11 0	10 0	0.42^*
淋巴结情况	N1	3	2	0.555	3	2	0.619	1	4	0.42^*
转移情况	M0	4	11 0	0.228	10 0	5	0.047*	10 0	5	0.021^*
转移情况	M1	6	5	0.228	3	8	0.047*	2	9	0.021^*
Gleason评分	3+4=7	2	4	0.702	1	5	0.129	5	1	0.047^*
Gleason评分	4+3=7	8	110	0.702	120	7	0.129	7	120	0.047^*

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Circulating tumor cells undergoing emt or expressing MAP1S help diagnosis of distant metastasis of prostate cancer

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Circulating tumor cells undergoing emt or expressing MAP1S help diagnosis of distant metastasis of prostate cancer