Objective To explore the effect of G-protein coupled receptor-associated sorting protein 1-short-hairpin RNA (GASP-1-ShRNA) on proliferation and apoptosis of lung cancer cells and tumor formation in nude mice.
Methods GASP-1-shRNA1, GASP-1-shRNA2 and GASP-1-shRNA3 were transfected into lung cancer A549 cells with Lipofectamine 2 000. Colony formation test and flow cytometry were used to detect proliferation and apoptosis of A549 cells from groups of GASP-1-shRNA1 transfection, blank control, and empty vector transfection, respectively. The A549 cells′ expressions of Ki67, cleaved caspase-3, B cell lymphoma-2 protein (Bcl-2) and Bcl-2-associated X protein (Bax) were determined with Western blot. An orthotopic xenograft mouse model was established through injection of GASP-1-shRNA1-transfected cells into hind leg tissues in nude mice. The tumor growth of the model mice were observed and the expressions of ki67 and caspase-3 protein in tumor tissues were detected with immunohistochemistry.
Results The mRNA and protein expression of GASP-1 in A549 cells were down-regulated after transfections of GASP-1-shRNA1, GASP-1-shRNA2, and GASP-1-shRNA3, especially in the GASP-1-shRNA1 transfected A549 cells. The down-regulation of GASP-1 could inhibit proliferation and promote cell apoptosis of the A549 cells. In vivo experiments, down-regulated GASP-1 could restrain tumor growth by inhibiting proliferation and promoting apoptosis of the A549 cells.
Conclusion Down-regulation of GASP-1 could inhibit proliferation and promote apoptosis of lung cancer A549 cells.
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