Adenovirus Vector-mediated Gene Transfer into Human Trabecular Cell.

Kee, Changwon; Son, Seongsu; Kim, Taeyoun; Lim, Jeomhee

Original Article

Journal of the Korean Ophthalmological Society 1999;40(5):1311-1317.
Published online May 1, 1999.

Adenovirus Vector-mediated Gene Transfer into Human Trabecular Cell.

Changwon Kee, Seongsu Son, Taeyoun Kim, Jeomhee Lim

Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University.

아데노바이러스 벡터를 이용한 섬유주 세포로의 유전자 전이

기창원(Chang Won Kee),손성수(Seong Su Son),김태윤(Tae Youn Kim),임점희(Jeom Hee Lim)

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.

Key Words: Gene therapy;Glaucoma;Recombinant adenovirus vector;Stromelysin cDNA