Effect of Galectin-3 Gene Transfer for the Apoptosis and Cell Growth of the Prostate Cancer Cell Line (LNCaP Cells) / 대한비뇨기과학회지

ABSTRACT

PURPOSE: Galectin-3 is a member of a large family of beta-galactoside- binding animal lectins. It is thought that galectin-3 can be a suppressor of apoptosis because of its significant sequence similarity with Bcl-2. We examined the role of galectin-3 for the paclitaxel-induced apoptosis after transfection of the galectin-3 gene in LNCaP cells. MATERIALS AND METHODS: Galectin-3 cDNA was cloned into PcDNA 3.1(-) and transfected into LNCaP cells. Stable transfection of galectin-3 into the LNCaP cells was achieved. Growth of the transfectants was observed with performing MTT assay. Apoptosis was induced by 100nM paclitaxel and 2microM staurosporine, and this was observed by DNA fragmentation assay. The viable cell numbers(% of control) after induction of apoptosis were determined with performing MTT assay. RESULTS: The LNCaP subclone that expressed galectin-3(LNCaP-G3-PcDNA) grew faster than the control transfectant(LNCaP-PcDNA)(p<0.05). The DNA fragmentation bands were decreased in the LNCaP subclone expressing galectin-3(LNCaP-G3-PcDNA) as compared to the control transfectant(LNCaP-PcDNA) after induction of apoptosis by 100nM paclitaxel or 2iM staurosporine; this means that galectin-3 inhibits apoptosis. The viable cells (% of control) with LNCaP-G3-PcDNA after the induction of apoptosis by 100nM paclitaxel was 92+/-2% in 8 hours, 77.5+/-1.9% in 24 hours and 40.4+/-2.9% in 48 hours on average, respectively. In contrast, the viable cells(% of control) of the control transfectant were 84.5+/-2%, 46+/-2.5% and 19+/-2.6% on average, respectively. The viable cells(% of control) with the LNCaP-G3-PcDNA after the induction of apoptosis by 100nM paclitaxel was higher than that of the control transfectant(LNCaP- PcDNA cells)(p<0.05). CONCLUSIONS: Galectin-3 gene transfer stimulates the growth of LNCaP cells. The galectin-3 protects LNCaP cells from paclitaxel-induced apoptosis.