Preparation of HSA-PEI-pcDNA-Apoptin and its effect on the proliferation of breast cancer MCF-7 cells

Liu Yi-zhi; Zhao Xin-han; Lv Wei-dong; Chang Bai-ling

doi:10.3969/j.issn.2095-4344.2016.49.019

Abstract：BACKGROUND:Apoptin is known to induce apoptosis of more than 50 kinds of tumor cel s. However, efficient systems are required to deliver apoptin to the cancer cel s for clinical use. <br> OBJECTIVE:To construct human serum albumin (HSA) and Apoptin complex and transfect it to cancer cel s in vitro and in vivo so as to find an efficient approach for apoptin delivery. <br> METHODS:Polyethylenimine was used to react with N-hydroxysuccinimide (NHS)-HSA solution to synthesized HSA-PEI, and then react with pcDNA-Apoptin to construct recombinant HSA-PEI-pcDNA-Apoptin. HSA-PEI-pcDNA-Apoptin complex was transformed into MCF-7 breast cancer cel lines. Then the expression of apoptin was detected by western blot assay and the cel proliferation was detected by MTT assay and flow cytometry. The MCF-7 breast cancer cel xenograft model was used to detect the in vivo performance of HSA-PEI-pcDNA-Apoptin by measuring the tumor volume at 4 weeks, with saline and HSA-PEI-pcDNA as controls. <br> RESULTS AND CONCLUSION:(1) We successful y prepared and confirmed the construction of HSA-PEI-pcDNA-Apoptin complex, which was successful y transformed into MCF-7 cel s. (2) MCF-7 cel s could express apoptin in the HSA-PEI-pcDNA-Apoptin group at 24 hours, but neither HSA-PEI-pcDNA group nor blank control group could express apoptin. (3) MTT assay for cel viability showed that HSA-PEI-pcDNA-Apoptin group had significantly lower optical density value than that in the other two groups (P<0.05). (4) The tumor volume in the HSA-PEI-pcDNA-Apoptin group was significantly less than that in the other two groups (P<0.05). (5) These findings indicate that the HSA-PEI-pcDNA-Apoptin complex markedly inhibits tumor growth in vitro and in vivo.