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[The construction and expression of a hepatocellular carcinoma-specific superantigen SEA (D227A) expression vector]
Shao-Ying Lu1, Yan-Fang Sui, Zeng-Shan Li
1Department of General Surgery, First Hospital of Xi'an Jiaotong University, Xi'an 710061 China.
Aim:
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.
Conclusion:
The successful construction and expression of hepatocellular carcinoma-specific superantigen SEA (D227A) gene expression vector lay the foundation for enhancing immunotherapy of hepatocellular carcinoma.
