Zhang Li-Wang, Ren Jun, Zhang Liang, Zhang Hong-Mei, Jin Bin, Pan Bo-Rong, Si Xiao-Ming, Zhang Yan-Jun, Wang Zhong-Hua, Pan Yang-Lin, Festein Stephen M
Department of Oncology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
World J Gastroenterol. 2003 Jul;9(7):1465-8. doi: 10.3748/wjg.v9.i7.1465.
To construct a recombined human AFP eukaryotic expression vector for the purpose of gene therapy and target therapy of hepatocellular carcinoma (HCC).
The full length AFP-cDNA of prokaryotic vector was digested, and subcloned to the multi-clony sites of the eukaryotic vector. The constructed vector was confirmed by enzymes digestion and electrophoresis, and the product expressed was detected by electrochemiluminescence and immunofluorescence methods.
The full length AFP-cDNA successfully cloned to the eukaryotic vector through electrophoresis, 0.9723 IU/ml AFP antigen was detected in the supernatant of AFP-CHO by electrochemiluminescence method. Compared with the control groups, the differences were significant (P<0.05). AFP antigen molecule was observed in the plasma of AFP-CHO by immunofluorescence staining.
The recombined human AFP eukaryotic expression vector can express in CHO cell line. It provides experimental data for gene therapy and target therapy of hepatocellular carcinoma.
构建重组人甲胎蛋白真核表达载体，用于肝细胞癌（HCC）的基因治疗和靶向治疗。
对原核载体的全长甲胎蛋白-cDNA进行酶切，并亚克隆至真核载体的多克隆位点。通过酶切和电泳对构建的载体进行鉴定，采用电化学发光和免疫荧光方法检测表达产物。
通过电泳成功将全长甲胎蛋白-cDNA克隆至真核载体，采用电化学发光法在甲胎蛋白-中国仓鼠卵巢细胞（AFP-CHO）的上清液中检测到0.9723 IU/ml的甲胎蛋白抗原。与对照组相比，差异具有统计学意义（P<0.05）。通过免疫荧光染色在AFP-CHO的细胞浆中观察到甲胎蛋白抗原分子。
重组人甲胎蛋白真核表达载体可在中国仓鼠卵巢细胞系中表达。为肝细胞癌的基因治疗和靶向治疗提供了实验数据。
