[The construction and expression of a hepatocellular carcinoma-specific superantigen SEA (D227A) expression vector].
OBJECTIVE
To construct the superantigen SEA (D227A) expression vector modulated by cis-acting element of AFP and express it specifically on AFP(+) hepatocellular carcinoma (HCC) cells.


METHODS
The enhancer and promoter of AFP gene, SEA (D227A) cDNA and linker-CD80tm were amplified by PCR. The gene segments mentioned above were cloned into the multiple cloning sites of retrovirus vector pLXSN to construct an attenuated superantigen expression vector pLXSN-SEA (D227A)-linker-CD80tm modulated by cis-acting element of AFP gene. AFP(+) or AFP(-) HCCs were transfected by the superantigen expression vector through lipofectamine mediation. SEA expression was detected by RT-PCR and indirect immunofluorescence staining.


RESULTS
Promoter, enhancer, SEA (D227A) and linker-CD80tm had been successfully cloned into the multiple cloning site of retrovirus vector pLXSN RT-PCR and indirect immunofluorescence assay were used to prove that SEA (D227A) could specifically express in the AFP(+) HCCs.


CONCLUSIONS
The successful construction and expression of hepatocellular carcinoma-specific superantigen SEA (D227A) gene expression vector lay the foundation for enhancing immunotherapy of hepatocellular carcinoma.
Lu
Shao-Ying
SY
Department of General Surgery, First Hospital of Xi'an Jiaotong University, Xi'an 710061 China.
Sui
Yan-Fang
YF
Li
Zeng-Shan
ZS
Pan
Cheng-En
CE
Ye
Jing
J
Zhang
Xiu-Min
XM
Wang
Wen-Yong
WY
D006528:Carcinoma, Hepatocellular / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D018089:Superantigens / Q000276:immunology; D014162:Transfection; D000509:alpha-Fetoproteins / Q000235:genetics
D018089:Superantigens; D000509:alpha-Fetoproteins