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

doi:10.3969/j.issn.2095-4344.2016.49.019

Abstract

Abstract:

BACKGROUND: Apoptin is known to induce apoptosis of more than 50 kinds of tumor cells. However, efficient systems are required to deliver apoptin to the cancer cells for clinical use.
OBJECTIVE: To construct human serum albumin (HSA) and Apoptin complex and transfect it to cancer cells in vitro and in vivo so as to find an efficient approach for apoptin delivery.
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 cell lines. Then the expression of apoptin was detected by western blot assay and the cell proliferation was detected by MTT assay and flow cytometry. The MCF-7 breast cancer cell 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.
RESULTS AND CONCLUSION: (1) We successfully prepared and confirmed the construction of HSA-PEI-pcDNA-Apoptin complex, which was successfully transformed into MCF-7 cells. (2) MCF-7 cells 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 cell 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.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Breast Neoplasms, Apoptosis, Gene Therapy, Tissue Engineering

CLC Number:

R318

Liu Yi-zhi, Zhao Xin-han, Lv Wei-dong, Chang Bai-ling . Preparation of HSA-PEI-pcDNA-Apoptin and its effect on the proliferation of breast cancer MCF-7 cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(49): 7425-7435.

Figures/Tables 5

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