Inhibition of semaphorin 4D enhances chemosensitivity by increasing 5-fluorouracile-induced apoptosis in colorectal canc
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ORIGINAL ARTICLE
Inhibition of semaphorin 4D enhances chemosensitivity by increasing 5-fluorouracile-induced apoptosis in colorectal cancer cells Golnaz Rashidi1 · Mahsa Rezaeepoor1 · Chiman Mohammadi2 · Ghasem Solgi1 · Rezvan Najafi2,3 Received: 7 May 2020 / Revised: 28 July 2020 / Accepted: 28 August 2020 © Springer Nature B.V. 2020
Abstract Overexpression of semaphorin 4D (SEMA4D), an immune semaphorin, is found in various human malignancies, including colorectal cancer (CRC). In this study, we explored the relationship between silencing SEMA4D expression and 5-fluorouracil (5-FU) response in the colorectal cancer cell line. SW48 cells were transfected with a short interfering RNA (siRNA) in order to silence SEMA4D gene expression and then exposed to 5-FU for 48 h. The down-regulation of SEMA4D expression was confirmed by qRT-PCR and the particular concentration of 5-FU was acquired using MTT assay. Flow cytometry and western blot were used to evaluate apoptosis rate and pro- and anti-apoptotic expression levels of proteins involved in apoptosis including Bax, Bcl-2, P53, and caspase-3. Other oncogenic activities including epithelial-mesenchymal transition (EMT) process, cancer stem cell (CSC) markers, and β-catenin pathway were investigated using qRT-PCR, and western blot. The proliferation was analyzed via colony formation test and cell invasion was assessed by transwell assay. Our data demonstrate that SEMA4D silencing results in strikingly elevated apoptosis in response to 5-FU treatment and leads to down-regulation of Bcl-2 and overexpression of Bax, P53, and caspase-3 in protein levels. Furthermore, the mRNA and protein expression levels of β-catenin, as well as transcript expressions of CSCs and EMT markers, were remarkably diminished. However, mRNA expression of E-cadherin as an epithelial marker was significantly increased in 5-FU treatment combined with siRNA SEMA4D. This study implicates that the silencing of SEMA4D by siRNA promotes the chemosensitivity of SW48 cells to 5-FU and it may be a potential therapeutic agent for colon cancer therapy. Keywords Colorectal cancer · SEMA4D · 5-FU · Apoptosis · EMT · CSCs
Introduction Colorectal cancer (CRC) is the second main factor of cancerrelated mortality, which ranks second in women and third in men [1]. Surgery, chemotherapy, and radiotherapy are the main therapeutic interventions in patients with colon cancer. Golnaz Rashidi and Mahsa Rezaeepoor have contributed equally to this work. * Rezvan Najafi [email protected]; [email protected] 1
Department of Immunology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
2
Department of Molecular Medicine and Genetics, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
3
Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
Despite the progression in colon cancer treatment, it is still a main public health issue all over the world [2]. Fluorouracil (5-FU) is a chemotherapeutic drug that is extensivel
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