Abstract
Targeted immunotherapy has become a popular research topic in cancer. The development and metastasis of cervical carcinoma are closely related to epidermal growth factor (EGF) and EGF-1 receptor (EGFR). We successfully constructed a single-chain human anti-EGFR antibody (scFv) and truncated protamine (tP) fusion protein (scFV/tP) expression vector using overlap extension PCR. Enzyme-linked immunosorbent assay and gel shift assay showed that the fusion protein retained the DNA and antigen-binding activity of the original antibody. Using the non-viral scFv/tP vector as a delivery tool, small interfering RNA (siRNA) of the human wings apart-like gene (hWAPL) was effectively transfected into cervical cancer HeLa cells. The hWAPL mRNA expression levels were reduced by 97.23 % in contrast with control cells, and the proliferation capability declined by 66.71 %, indicating significant inhibition. The present results provide a novel strategy for targeted gene therapy and siRNA therapy of EGFR-positive cervical cancers.
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Acknowledgments
This work was supported by grants from the Medical Science and Technique Development Fund, Nanjing, Jiangsu, China (No. ZKX09015), the Nanjing Municipal Science and Technology Commission, Jiangsu, China (No. 20120825), the Medical Science and Technique Development Fund, Nanjing, Jiangsu, China (No. ZKX12025) and the National Natural Science Foundation for Youth of China, China (No. 81301951).
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The authors declare that they have no competing interests.
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Huilin Zhang and Yuan Mao contributed equally to this work.
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Zhang, H., Mao, Y., Zhang, F. et al. The inhibitory effect of a new scFv/tP protein as siRNA delivery system to target hWAPL in cervical carcinoma. Mol Cell Biochem 391, 77–84 (2014). https://doi.org/10.1007/s11010-014-1989-3
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DOI: https://doi.org/10.1007/s11010-014-1989-3