Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Biochim Biophys Acta Mol Cell Res. 2022 May;1869(5):119221. doi: 10.1016/j.bbamcr.2022.119221. Epub 2022 Feb 5.
Prostate cancer continues to be one of the main global health issues in men. Neuropeptide substance P (SP) acting via neurokinin-1receptor (NK1R) promotes tumorigenicity in many human malignant tumors. However, its pro-tumorigenic functions and the therapeutic effects of its inhibition in prostate cancer remain unclear.
MTT assay was employed for measuring cellular proliferation and cytotoxicity. mRNAs and proteins expression levels were evaluated by qRT-PCR and western blot assay, respectively. Gelatinase activity was assessed by zymography. The migration ability was defined using wound-healing assay. Flow cytometry was employed to evaluate the cell cycle distribution. We also performed an in vivo experiment in a mouse model of prostate cancer to confirm the in vitro therapeutic effect of targeting the SP/NK1R system.
We found a noticeable increase in the expression of the truncated isoform of NK1R as an oncogenic NK1R splice variant in tumor cells. We also demonstrated that SP promotes both proliferative and migrative phenotypes of prostate cancer through modifying cell cycle-related proteins (c-Myc, cyclin D1, cyclin B1, p21), and apoptosis-related genes (Bcl-2 and Bax), promoting cell migration and increasing MMP-2 and MMP-9 expression and activity, while aprepitant administration could remarkably reverse these effects. SP also stimulated tumor growth in vivo, which was correlated with shorter survival times, while aprepitant reversed this effect and led to significantly longer survival time.
Our findings suggest that SP/NK1R system may serve as a novel therapeutic target in prostate cancer and support the possible candidacy of aprepitant in future prostate cancer therapy.
前列腺癌仍是男性面临的主要全球健康问题之一。神经肽 P 物质（SP）通过神经激肽-1 受体（NK1R）发挥作用，可促进多种人类恶性肿瘤的致瘤性。然而，其促肿瘤发生功能及其在前列腺癌中的抑制作用仍不清楚。
采用 MTT 法检测细胞增殖和细胞毒性。分别采用 qRT-PCR 和 Western blot 法检测 mRNAs 和蛋白质表达水平。通过明胶酶谱法评估凝胶酶活性。采用划痕愈合实验评估迁移能力。采用流式细胞术评估细胞周期分布。我们还在前列腺癌小鼠模型中进行了体内实验，以证实靶向 SP/NK1R 系统的体外治疗效果。
我们发现肿瘤细胞中存在截断型 NK1R 异构体的表达显著增加，这是一种致癌性 NK1R 剪接变体。我们还表明，SP 通过调节细胞周期相关蛋白（c-Myc、cyclin D1、cyclin B1、p21）和凋亡相关基因（Bcl-2 和 Bax）促进前列腺癌细胞的增殖和迁移表型，促进细胞迁移，增加 MMP-2 和 MMP-9 的表达和活性，而阿瑞匹坦给药可显著逆转这些作用。SP 还可刺激体内肿瘤生长，与较短的生存时间相关，而阿瑞匹坦可逆转这一作用并导致显著延长的生存时间。
我们的研究结果表明，SP/NK1R 系统可能成为前列腺癌的一种新的治疗靶点，并支持阿瑞匹坦在未来前列腺癌治疗中的候选地位。
