Additive cytotoxic effect of apoptin and chemotherapeutic agent docetaxel on lncap cell lines

Vida Mohammadi,1,* Mohammad ali takhshid,2 Golamreza rafiei dehbidi ,3

1. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
2. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
3. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Introduction

Unfortunately although chemotherapy is primary therapy for most types of advanced prostate cancer, conventional chemotherapeutic agents are not effective for advanced prostate cancer. the effectiveness of combination of chemotherapy and gene therapy in the cancer therapy era reported from different studies. apoptin (vp3) is a viral protein with 121 residues from chicken anemia virus (cav) that revealed significant apoptotic activity in various tumor cell lines. in this study, we evaluated the apoptosis effect of apoptin overexpression in combination with docetaxel as chemotherapy agent on the lncap cells.

Methods

Docetaxcel concentration required for inhibiting 50% of lncap cells (ic50) were determined in different concentration of docedoxel (1nm to 100 nm) by mtt assay. the expression plasmid (adeno shuttle vector of pad-cmv) contains apoptin under the regulation of the cytomegalovirus (cmv) promoter was constructed. in order to investige the synergistic effect of apoptin overexpression in combined with doxetaxel, three groups of lncap cells were seeded in 6 well plates. after 75-90% confluency, a group of these cells trasfected with adeno shuttle vector of pad- cmv-apoptin, another group was transfected only with docetaxel and last group treated with combination of docetaxel and apoptin. fourty-eight hours after treatment, synergistic effect of these combination on apoptosis was measured using annexin v-7aad flow cytometric method and results compared with the group treated with adeno shuttle vector of pad- cmv-apoptin only.

Results

Flowcytometry assay showed that combined pad-cmv-apoptin/docetaxel treatment significantly increased the antitumor effect in comparison to the single treatments.

Conclusion

In conclusion, our study indicate docetaxel treatment combined with apoptin expression resulted in an additive cytotoxic activity in lncap cells.

Keywords

Combine therapy, apoptin, docetaxel, apoptosis