Topcu Kubra Sena Bas, Korucu Emine Nedime, Menevse Esma, Kocak Nadir, Duran Tugce
Department of Molecular Biology and Genetics, Faculty of Science, Bartın University, Bartın, Turkey.
Department of Molecular Biology and Genetics, Faculty of Science, Necmettin Erbakan University, Konya, Turkey.
Invest New Drugs. 2022 Jun;40(3):519-528. doi: 10.1007/s10637-021-01209-z. Epub 2022 Feb 3.
Pancreatic ductal adenocarcinoma (PDAC) is one of the most common malignant tumors of the pancreas. Preclinical studies show that it evades the immune system with immune checkpoints and promotes tumor development. V-domain Ig suppressor of T cell activation (VISTA) is a new immune-check point from the B7 family and is highly expressed in cancer cells. Overexpression of toll like receptor 4 (TLR4) in pancreatic adenocarcinoma is associated with induced tumorigenesis, tumor growth, resistancy to chemotherapy. Naloxone is an opioid and inhibits TLR4-ligand association. In this study, we investigated the relation of TLR4 and downstream pathways with immune-check point VISTA in pancreatic cancer proliferation. We initially collected pancreatic cancer-related datasets using the GEPIA2 and UALCAN databases. Based on this data obtained the effect of various concentrations and incubation times of naloxone were used on PANC-1 cells proliferation. A combination of naloxone and VISTA-siRNA were applied, and the effect of both naloxone and combined treatment on TLR4, Interleukin 1 receptor associated kinase 4 (IRAK4) and VISTA gene expression were analyzed in pancreatic cancer cells. As a result of analysis with Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), gene expression levels of TLR4, IRAK4 and VISTA were significantly suppressed and cell proliferation was significantly reduced. We found that administration of naloxone and VISTA-siRNA in combination with PDAC cells suppressed signaling. Therefore, we considered that the relationship between VISTA and TLR4 signaling pathways and the other possible associated signal molecules may be an important marker in determining the response of immune checkpoint inhibitors in cancer treatment.
胰腺导管腺癌（PDAC）是胰腺最常见的恶性肿瘤之一。临床前研究表明，它通过免疫检查点逃避免疫系统并促进肿瘤发展。T细胞激活的V结构域Ig抑制因子（VISTA）是B7家族的一个新的免疫检查点，在癌细胞中高表达。胰腺腺癌中Toll样受体4（TLR4）的过表达与肿瘤发生、肿瘤生长、化疗耐药相关。纳洛酮是一种阿片类药物，可抑制TLR4与配体的结合。在本研究中，我们调查了TLR4及其下游通路与免疫检查点VISTA在胰腺癌增殖中的关系。我们最初使用GEPIA2和UALCAN数据库收集了胰腺癌相关数据集。基于这些数据，研究了不同浓度和孵育时间的纳洛酮对PANC-1细胞增殖的影响。应用纳洛酮和VISTA-siRNA的组合，并分析了纳洛酮及联合治疗对胰腺癌细胞中TLR4、白介素1受体相关激酶4（IRAK4）和VISTA基因表达的影响。定量实时聚合酶链反应（qRT-PCR）分析结果显示，TLR4、IRAK4和VISTA的基因表达水平显著受到抑制，细胞增殖也显著降低。我们发现，将纳洛酮和VISTA-siRNA与胰腺导管腺癌细胞联合使用可抑制信号传导。因此，我们认为VISTA与TLR4信号通路以及其他可能相关的信号分子之间的关系可能是确定癌症治疗中免疫检查点抑制剂反应的一个重要标志物。
