Document Type : Original Article

Authors

• Morteza Akbari ^{1, 2, 3}
• Dariush Shanehbandi ²
• Milad Asadi ²
• Navid Shomali ^{2, 4}
• Afsaneh Faraji ²
• Vahid Khaze ²
• Abbas Pakdel ⁵
• Ahad Mokhtarzadeh ²
• Ali Asghar Ebrahimi ⁶
• Aliakbar Shabani ^{1, 4}
• Behzad Baradaran ²

¹ Department of Biotechnology, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran

² Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

³ Semnan Biotechnology Research Center, Semnan University of Medical sciences, Semnan, Iran

⁴ Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

⁵ Department of Biochemistry, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran

⁶ Department of Rheumatology, Tabriz University of Medical Sciences, Tabriz, Iran

Abstract

Background: Colorectal cancer (CRC) is attributed as one of the most common malignancies worldwide. CD133 molecule, as a pentaspan transmembrane glycoprotein, confers stem cell-related characteristics, including self-renewal and multi-directional differentiation capability. CD133 plays important roles in the progression of CRC by conferring apoptotic resistance and migration ability. Objective: To investigate the anti-apoptotic and anti-angiogenic effect of CD-133 targeted siRNA in a colorectal cancer cell line. Methods: In this study, CD133-targeted siRNA transfection was conducted into HT-29 cells. MTT assay was employed to evaluate the cytotoxic effects of transfection on the cells. Flow cytometry was used to evaluate the apoptosis rate. The mRNA expression of apoptosis and metastasis related genes were assessed by quantitative Real-Time PCR (qRT-PCR). Wound healing assay was used to assess the migration potency of the infected cells. Results: Expression of CD133 was significantly downregulated after transfection of CD133-specific siRNA. Moreover, the rate of apoptosis was significantly increased after transfection. The migration potential of cells was diminished after transfection. siRNA delivery resulted in the modulation of expression of apoptosis and metastasis-related genes. Conclusion: siRNA mediated targeting of CD133 could be considered as a promising approach to treat CRC through suppressing the cancerous behavior of tumor cells.

Keywords

• Apoptosis
• Colorectal Cancer
• CD133
• Metastasis
• siRNA