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优化流式细胞术检测人血液系统恶性肿瘤中瞬时受体电位香草酸 1 型(TRPV1)。

Optimized flow cytometric detection of transient receptor potential vanilloid-1 (TRPV1) in human hematological malignancies.

机构信息

School of Health Sciences, University of Tasmania, Locked Bag 1322, Launceston, TAS, 7250, Australia.

Children's Cancer Institute, School of Women's and Children's Health, Lowy Cancer Centre, University of New South Wales (UNSW), Sydney, NSW, Australia.

出版信息

Med Oncol. 2022 Apr 28;39(5):81. doi: 10.1007/s12032-022-01678-z.

Abstract

The ectopic overexpression of transient receptor potential vanilloid-1 (TRPV1) has been detected in numerous solid cancers, including breast, prostate, pancreatic, and tongue epithelium cancer. However, the expression of TRPV1 in hematological malignancies remains unknown. Here we show through in silico analysis that elevated TRPV1 mRNA expression occurs in a range of hematological malignancies and presents an optimized flow cytometry method to rapidly assess TRPV1 protein expression for both cell lines and primary patient samples. Three anti-TRPV1 antibodies were evaluated for intracellular TRPV1 detection using flow cytometry resulting in an optimized protocol for the evaluation of TRPV1 in hematological malignant cell lines and patients' peripheral blood mononuclear cells (PBMC). Overexpression of TRPV1 was observed in THP-1 (acute monocytic leukemia) and U266B1 (multiple myeloma, MM), but not U937 (histiocytic lymphoma) compared to healthy PBMC. TRPV1 was also detected in all 49 patients including B-cell non-Hodgkin's lymphoma (B-NHL), MM, and others and 20 healthy controls. TRPV1 expression was increased in 8% of patients (MM = 2, B-NHL = 2). In conclusion, we provide an optimized flow cytometry method for routine expression analysis of clinical samples and show that TRPV1 is increased in a subset of patients with hematological malignancies.

摘要

瞬时受体电位香草酸 1(TRPV1)的异位过表达已在许多实体瘤中被检测到,包括乳腺癌、前列腺癌、胰腺癌和舌上皮癌。然而,TRPV1 在血液恶性肿瘤中的表达尚不清楚。在这里,我们通过计算机分析表明,一系列血液恶性肿瘤中存在 TRPV1 mRNA 表达升高,并提出了一种优化的流式细胞术方法,用于快速评估细胞系和原发性患者样本中的 TRPV1 蛋白表达。使用流式细胞术评估了三种抗 TRPV1 抗体对细胞内 TRPV1 的检测,从而优化了评估血液恶性肿瘤细胞系和患者外周血单核细胞(PBMC)中 TRPV1 的方案。与健康 PBMC 相比,在 THP-1(急性单核细胞白血病)和 U266B1(多发性骨髓瘤,MM)中观察到 TRPV1 的过表达,但在 U937(组织细胞淋巴瘤)中未观察到。TRPV1 也在包括 B 细胞非霍奇金淋巴瘤(B-NHL)、MM 和其他患者以及 20 名健康对照者在内的所有 49 名患者中被检测到。TRPV1 表达在 8%的患者中增加(MM=2,B-NHL=2)。总之,我们提供了一种优化的流式细胞术方法,用于对临床样本进行常规表达分析,并表明 TRPV1 在血液恶性肿瘤患者的亚组中增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/62b1/9046313/fc461e4abfcf/12032_2022_1678_Fig1_HTML.jpg

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