title: The inhibitory effect of a new scFv/tP protein as siRNA delivery system to target hWAPL in cervical carcinoma.
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.
authors.affiliation: Department of Gynaecology and Obstetrics, Nanjing Maternal and Children Care Hospital Affiliated to Nanjing Medical University, 210004, Nanjing, China.
authors.forename: Huilin
authors.initials: H
authors.lastname: Zhang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Mao
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.forename: Chunping
authors.initials: C
authors.lastname: Ye
authors.forename: Hua
authors.initials: H
authors.lastname: Tong
authors.forename: Yiping
authors.initials: Y
authors.lastname: Su
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000941:Antigens / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011479:Protamines / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000378:metabolism*; D014162:Transfection; D002583:Uterine Cervical Neoplasms / Q000378:metabolism* / Q000473:pathology; D000097722:Cohesins
chemical_list: D000941:Antigens; D002352:Carrier Proteins; D004247:DNA; D009687:Nuclear Proteins; D011479:Protamines; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C488092:WAPL protein, human; D000097722:Cohesins