Zhang Huilin, Mao Yuan, Zhang Feng, Ye Chunping, Tong Hua, Su Yiping, Zhu Jin
Department of Gynaecology and Obstetrics, Nanjing Maternal and Children Care Hospital Affiliated to Nanjing Medical University, 210004, Nanjing, China.
Mol Cell Biochem. 2014 Jun;391(1-2):77-84. doi: 10.1007/s11010-014-1989-3. Epub 2014 Feb 25.
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.
靶向免疫疗法已成为癌症领域一个热门的研究课题。宫颈癌的发生发展及转移与表皮生长因子（EGF）和表皮生长因子-1受体（EGFR）密切相关。我们利用重叠延伸PCR成功构建了单链人抗EGFR抗体（scFv）与截短的鱼精蛋白（tP）融合蛋白（scFV/tP）表达载体。酶联免疫吸附测定和凝胶迁移试验表明，该融合蛋白保留了原始抗体的DNA结合活性和抗原结合活性。以非病毒scFv/tP载体作为递送工具，将人分离酶抑制因子基因（hWAPL）的小干扰RNA（siRNA）有效转染至宫颈癌HeLa细胞中。与对照细胞相比，hWAPL mRNA表达水平降低了97.23%，增殖能力下降了66.71%，表明有显著抑制作用。目前的研究结果为EGFR阳性宫颈癌的靶向基因治疗和siRNA治疗提供了一种新策略。
