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[Molecular cloning of a novel human lung cancer-associated antigen cDNA]
1Department of Biochemistry & Molecular Biology, China-Japan Friendship Institute of Clinical Medical Sciences, Beijing.
Objective:
To clone the gene of human lung cancer-associated antigen.
Methods:
The human lung squamous carcinoma cell line L-78 cDNA expression library was constructed with lambda gt 11 Sfi-Not directional cloning vector and screened with the anti-human lung cancer McAb ALT-04. The cDNA clone named hlc-14 was sequenced and further studied. NIH3T3 cells were transfected with the recombinant pXJ41-hlc14 plasmid, and the expression of hlc-14 cDNA was detected and characterized immunohistochemically. Soft agar colony formation assay of the transfectants was performed.
Results:
A human lung cancer lambda gt 11 cDNA library of 1.4 x 10(6) pfu/ml was constructed. Six cDNA clones were isolated following 4 rounds of immunoscreening. The largest cDNA clone hlc-14 contained 954 bp with no homology to any known gene when screened through GenBank. In the NIH3T3 cells transfectant of hlc-14 cDNA, the expression of the lung cancer-associated antigen was demonstrated. The ability of colony formation of the transfectant in the soft agar was enhanced.
Conclusion:
The hlc-14 clone is a novel human lung cancer-associated antigen cDNA. It provides clues for the study of carcinogenesis mechanism of lung cancer and a novel target gene for the gene therapy of lung cancer.