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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens / metabolism
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Cell Death / drug effects
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Cell Proliferation / drug effects
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DNA / metabolism
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Electrophoresis, Polyacrylamide Gel
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Electrophoretic Mobility Shift Assay
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Enzyme-Linked Immunosorbent Assay
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Female
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Gene Transfer Techniques*
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HeLa Cells
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Humans
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Protamines / metabolism*
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Protein Binding / drug effects
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism*
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RNA, Messenger / genetics
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RNA, Messenger / metabolism
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RNA, Small Interfering / metabolism*
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism*
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Recombinant Fusion Proteins / pharmacology
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Single-Chain Antibodies / metabolism*
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Transfection
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Uterine Cervical Neoplasms / metabolism*
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Uterine Cervical Neoplasms / pathology
Substances
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Antigens
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Carrier Proteins
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Nuclear Proteins
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Protamines
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Proto-Oncogene Proteins
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RNA, Messenger
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RNA, Small Interfering
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Recombinant Fusion Proteins
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Single-Chain Antibodies
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WAPL protein, human
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DNA