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^*

[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]	Tosoian JJ, Mamawala M, Epstein JI, et al. Intermediate and longer-term outcomes from a prospective active-surveillance program for favorable-risk prostate cancer[J]. J Clin Oncol, 2015, 33(30): 3379-85.
[3]	Bubendorf L, Schopfer A, Wagner U, et al. Metastatic patterns of prostate cancer: an autopsy study of 1,589 patients [J]. Hum Pathol, 2000, 31(5): 578-583.
[4]	Norgaard M, Jensen AO, Jacobsen JB, et al. Skeletal related events, bone metastasis and survival of prostate cancer: a population based cohort study in Denmark (1999 to 2007) [J]. J Urol, 2010, 184(1): 162-167.
[5]	Croucher PI, Mcdonald MM, Martin TJ. Bone metastasis: the importance of the neighbourhood [J]. Nat Rev Cancer, 2016, 16(6): 373-386.
[6]	Futakuchi M, Fukamachi K, Suzui M. Heterogeneity of tumor cells in the bone microenvironment: Mechanisms and therapeutic targets for bone metastasis of prostate or breast cancer[J]. Adv Drug Deliv Rev, 2016, 99(Pt B): 206-211.
[7]	Cortes-hernandez LE, Eslami SZ, Alix-panabieres C. Circulating tumor cell as the functional aspect of liquid biopsy to understand the metastatic cascade in solid cancer [J]. Mol Aspects Med, 2020, 72：100816.
[8]	Boerrigter E, Groen LN, Van erp NP, et al. Clinical utility of emerging biomarkers in prostate cancer liquid biopsies[J]. Expert Rev Mol Diagn, 2020, 20(2): 219-230.
[9]	Aceto N, Toner M, Maheswaran S, et al. En Route to Metastasis: Circulating Tumor Cell Clusters and Epithelial-to-Mesenchymal Transition [J]. Trends Cancer, 2015, 1(1): 44-52.
[10]	Mizukoshi K, Okazawa Y, Haeno H, et al. Metastatic seeding of human colon cancer cell clusters expressing the hybrid epithelial/mesenchymal state [J]. Int J Cancer, 2020, 146(9): 2547-2562.
[11]	Jolly MK, Mani SA, levine H. Hybrid epithelial/mesenchymal phenotype(s): The 'fittest' for metastasis? [J]. Biochimica et biophysica acta Reviews on cancer, 2018, 1870(2): 151-157.
[12]	Qi LN, Xiang BD, Wu FX, et al. Circulating tumor cells undergoing emt provide a metric for diagnosis and prognosis of patients with hepatocellular carcinoma [J]. Cancer Res, 2018, 78(16): 4731-4744.
[13]	Wang W, Wan L, Wu S, et al. Mesenchymal marker and LGR5 expression levels in circulating tumor cells correlate with colorectal cancer prognosis [J]. Cell Oncol (Dordr), 2018, 41(5): 495-504.
[14]	Li TT, Liu H, Li FP, et al. Evaluation of epithelial-mesenchymal transitioned circulating tumor cells in patients with resectable gastric cancer: Relevance to therapy response [J]. World J Gastroenterol, 2015, 21(47): 13259-13267.
[15]	Zhang S, Wu T, Peng X, et al. Mesenchymal phenotype of circulating tumor cells is associated with distant metastasis in breast cancer patients [J]. Cancer Manag Res, 2017, 9(691-700).
[16]	Jiang X, Zhong W, Huang H, et al. Autophagy defects suggested by low levels of autophagy activator MAP1S and high levels of autophagy inhibitor LRPPRC predict poor prognosis of prostate cancer patients [J]. Mol Carcinog, 2015, 54(10): 1194-1204.
[17]	Kiyono K, Suzuki HI, Matsuyama H, et al. Autophagy is activated by TGF-beta and potentiates TGF-beta-mediated growth inhibition in human hepatocellular carcinoma cells [J]. Cancer Res, 2009, 69(23): 8844-8852.
[18]	童行, 何卫阳. 自噬在肿瘤上皮-间充质转化以及侵袭转移中的作用 [J]. 现代泌尿外科杂志, 2019, 24(5): 81-85.
[19]	Amantini C, Morelli MB, Nabissi M, et al. Capsaicin triggers autophagic cell survival which drives epithelial mesenchymal transition and chemoresistance in bladder cancer cells in an Hedgehog-dependent manner [J]. Oncotarget, 2016, 7(31): 50180-50194.
[20]	Broncy L, Paterlini-brechot P. Clinical Impact of Circulating Tumor Cells in Patients with Localized Prostate Cancer [J]. Cells, 2019, 8(7): 676.
[21]	Zapatero A, Gomez-caamano A, Cabeza rodriguez MA, et al. Detection and dynamics of circulating tumor cells in patients with high-risk prostate cancer treated with radiotherapy and hormones: a prospective phase II study[J]. Radiat Oncol, 2020, 15(1): 137.
[22]	Liu DG, Xue L, Li J, et al. Epithelial-mesenchymal transition and GALC expression of circulating tumor cells indicate metastasis and poor prognosis in non-small cell lung cancer [J]. Cancer Biomark, 2018, 22(3): 417-426.
[23]	Liu L, Mckeehan WL, Wang F, et al. MAP1S enhances autophagy to suppress tumorigenesis[J]. Autophagy, 2012, 8(2): 278-280.
[24]	Gao C, Cao W, Bao L, et al. Autophagy negatively regulates Wnt signalling by promoting Dishevelled degradation [J]. Nat Cell Biol, 2010, 12(8): 781-790.
[25]	De francesco EM, Maggiolini M, Musti AM. Crosstalk between Notch, HIF-1alpha and GPER in Breast Cancer EMT [J]. Int J Mol Sci, 2018, 19(7): 2011.
[26]	Zhang PP, Wang PQ, Qiao CP, et al. Differentiation therapy of hepatocellular carcinoma by inhibiting the activity of AKT/GSK-3beta/beta-catenin axis and TGF-beta induced EMT with sophocarpine [J]. Cancer Lett, 2016, 376(1): 95-103.

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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