Department of Molecular Biology and Genetics, Gebze Technical University, Kocaeli 41400, Turkey.
Protein Core Facility, Cancer Research UK and Experimental Cancer Medicine Centres, University of Southampton, Southampton General Hospital, Southampton SO16 6YD, UK.
Cells. 2019 Mar 2;8(3):210. doi: 10.3390/cells8030210.
Hepatocellular carcinoma (HCC) is a common and deadly cancer; however, very little improvement has been made towards its diagnosis and prognosis. The expression and functional contribution of the receptor tyrosine kinase ROR1 have not been investigated in HCC before. Hence, we investigated the expression of ROR1 in HCC cells and assessed its involvement in hepatocarcinogenesis.
Recombinant bacterial ROR1 protein was used as an immunogen to generate ROR1 monoclonal antibodies. transcript levels were detected by RT-qPCR and the protein expression of ROR1 in HCC was assessed by Western blotting by using homemade anti-ROR1 monoclonal antibodies. Apoptosis, cell cycle, trans-well migration, and drug efflux assays were performed in shRNA-ROR1 HCC cell clones to uncover the functional contribution of ROR1 to hepatocarcinogenesis.
New ROR1 antibodies specifically detected endogenous ROR1 protein in human and mouse HCC cell lines. ROR1-knockdown resulted in decreased proliferation and migration but enhanced resistance to apoptosis and anoikis. The observed chemotherapy-resistant phenotype of ROR1-knockdown cells was due to enhanced drug efflux and increased expression of multi-drug resistance genes.
ROR1 is expressed in HCC and contributes to disease development by interfering with multiple pathways. Acquired ROR1 expression may have diagnostic and prognostic value in HCC.
肝细胞癌（HCC）是一种常见且致命的癌症；然而，在其诊断和预后方面几乎没有取得任何进展。受体酪氨酸激酶 ROR1 的表达及其在 HCC 中的功能贡献以前尚未在 HCC 中进行过研究。因此，我们研究了 ROR1 在 HCC 细胞中的表达，并评估了其在肝癌发生中的作用。
使用重组细菌 ROR1 蛋白作为免疫原生成 ROR1 单克隆抗体。通过 RT-qPCR 检测 转录水平，并使用自制的抗 ROR1 单克隆抗体通过 Western blot 评估 HCC 中 ROR1 的蛋白表达。在 shRNA-ROR1 HCC 细胞克隆中进行细胞凋亡、细胞周期、Transwell 迁移和药物外排测定，以揭示 ROR1 对肝癌发生的功能贡献。
新型 ROR1 抗体特异性检测了人源和鼠源 HCC 细胞系中的内源性 ROR1 蛋白。ROR1 敲低导致增殖和迁移减少，但增强了对细胞凋亡和失巢凋亡的抵抗。ROR1 敲低细胞观察到的化疗耐药表型是由于药物外排增强和多药耐药基因表达增加所致。
ROR1 在 HCC 中表达，并通过干扰多种途径促进疾病发展。获得性 ROR1 表达可能对 HCC 具有诊断和预后价值。
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