Document Type : Original Article

Authors

• Puria Rostampour ¹
• Jamileh Saberzadeh ¹
• Bahareh Zamani ¹
• Soofia Sorourian ²
• Mohammad Ali Takhshid ^{1, 2}

¹ Division of Medical Biotechnology, Department of Laboratory Sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

² Diagnostic Laboratory Sciences and Technology Research Centre, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Background: Chemoresistance is a significant obstacle that hinders the effectiveness of cisplatin. This study aimed to explore the potential effect of annexin A5 (ANXA5) on sensitizing HCT116 cells to the cytotoxic effects of cisplatin.
Methods: A recombinant pCMV6-ANXA5-IRES-GFP plasmid was utilized to overexpress ANXA5 in HCT116 cells. The plasmids were transfected using Lipofectamine 3000, and the ANXA5 expression was quantified using quantitative real-time PCR (q-PCR). The effects of ANXA5 overexpression on increasing the sensitivity of HCT116 cells to the cytotoxic effects of cisplatin on viability, apoptosis, and cell cycle were assessed using MTT assay, annexin V/7ADD flow cytometry, and cell cycle assay. The effects of the treatments on the expression of Bax and Bcl2 were measured using q-PCR.
Results: The overexpression of ANXA5 was confirmed by q-PCR (p<0.001). Cisplatin induced cell death, apoptosis, and cell cycle arrest. The effects of cisplatin on cell death (p = 0.026), apoptosis (p=0.001), and the Bax/Bcl2 ratio(p=0.032) were enhanced by ANXA5 overexpression. However, ANXA5 had no significant effect on the cell cycle arrest induced by cisplatin.
Conclusion: These findings suggest that ANXA5 can sensitize HCT116 cells to the cytotoxic effects of cisplatin.

Keywords

• Annexin A5
• Chemotherapy
• Colorectal cancer
• Apoptosis
• Cell cycle

Main Subjects

• Medical Biotechnology