Adenovirus Vector-mediated Gene Transfer into Human Trabecular Cell

ABSTRACT

We attempted to observe the expression of stromelysin, which regulates the extracellular matrix of trabecular meshwork and is one of the family of the matrix metalloproteinases, in the trabecular cells after transfection of replication deficient recombinant adenovirus vector containing stromelysin cDNA. Stromelysin cDNA was produced by RT-PCR with total RNA extracted from cultured human trabecular cells after induction with interleukin-1 alpha, and cloned by inserting the cDNA into the TA vector. Adenovirus vector that contains stromelysin cDNA was constructed by cotransfection of pJM17 and p delta A. CMV. PA-str into the 293 cells. The expression of stromelysin in the trabecular cells was assayed by Western blot and zymography. The sequence of stromelysin cDNA was consistent with that previously reported. The constructed adenovirus vector had stromelysin cDNA but had no E1 region. The expression of stromelysin in the trabecular cells by this vector was detected in 4 days and peaked in 7 days after transfection. In conclusion, this study showed the possibility of gene therapy in the glaucoma treatment by transfecting the trabecular cells with the replication deficient recombinant adenovirus vector containing stromelysin cDNA.