Construction and expression of recombined human AFP eukaryotic expression vector.
OBJECTIVE
To construct a recombined human AFP eukaryotic expression vector for the purpose of gene therapy and target therapy of hepatocellular carcinoma (HCC).


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Zhang
Li-Wang
LW
Department of Oncology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, China.
Ren
Jun
J
Zhang
Liang
L
Zhang
Hong-Mei
HM
Jin
Bin
B
Pan
Bo-Rong
BR
Si
Xiao-Ming
XM
Zhang
Yan-Jun
YJ
Wang
Zhong-Hua
ZH
Pan
Yang-Lin
YL
Festein
Stephen M
SM
D000818:Animals; D016466:CHO Cells; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000628:therapy*; D006224:Cricetinae; D018076:DNA, Complementary; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000628:therapy*; D011994:Recombinant Proteins / Q000235:genetics*; D014162:Transfection; D000509:alpha-Fetoproteins / Q000235:genetics*
D018076:DNA, Complementary; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins