Department of Immunology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Biomed Res Int. 2021 Jul 21;2021:8507373. doi: 10.1155/2021/8507373. eCollection 2021.
Semaphorin 4D (SEMA4D), a protein originally demonstrated to regulate the immune system and axonal growth cone collapse in the developing central nervous system, is overexpressed in various human malignancies, including colorectal cancer (CRC). This investigation was undertaken to examine the effects of SEMA4D silencing on the biological properties of the CRC cell line. SW48 cells were transfected with a siRNA-targeting SEMA4D. The mRNA expression of underlying pro- and antiapoptotic proteins including Bax, Bcl-2, P53, and caspase-3, cancer stem cell (CSC) markers, epithelial-mesenchymal transition (EMT) markers, MMP-2, and MMP-9 was examined using qRT-PCR. Further, the protein expression of E-cadherin and -catenin was confirmed by Western blot. SW48 cell migration and MMP activity were detected using scratch and zymography analysis, respectively. Finally, the apoptosis rate was assessed via the flowcytometry test. SEMA4D knock-down was associated with a considerable suppression of in vitro cell viability, EMT-related genes, CSC markers, -catenin signaling pathway, sphere-forming, cell migration, and MMP-2 activity as well as induction of apoptosis. This study identifies the inhibitory effects of SEMA4D gene silencing on tumor progression. Thereby, this might conclude a possible alternative to cancer therapy by targeting several prominent pathways involved in cancer through SEMA4D suppression.
信号素 4D（SEMA4D），一种最初被证明可调节免疫系统和中枢神经系统发育过程中轴突生长锥崩溃的蛋白质，在各种人类恶性肿瘤中过表达，包括结直肠癌（CRC）。本研究旨在研究 SEMA4D 沉默对 CRC 细胞系生物学特性的影响。用靶向 SEMA4D 的 siRNA 转染 SW48 细胞。使用 qRT-PCR 检测包括 Bax、Bcl-2、P53 和 caspase-3 在内的促凋亡和抗凋亡蛋白、癌症干细胞（CSC）标志物、上皮-间充质转化（EMT）标志物、MMP-2 和 MMP-9 的 mRNA 表达。此外，通过 Western blot 证实了 E-钙粘蛋白和 -连环蛋白的蛋白表达。通过划痕和明胶酶谱分析分别检测 SW48 细胞迁移和 MMP 活性。最后，通过流式细胞术试验评估细胞凋亡率。SEMA4D 敲低与体外细胞活力、EMT 相关基因、CSC 标志物、-连环蛋白信号通路、球体形成、细胞迁移和 MMP-2 活性的显著抑制以及细胞凋亡的诱导有关。本研究确定了 SEMA4D 基因沉默对肿瘤进展的抑制作用。因此，通过抑制 SEMA4D 可能会针对涉及癌症的几个重要途径，为癌症治疗提供一种替代方法。
