Inhibition of Proliferation of Human Osteosarcoma Cells Transfected with PIN1 Antisense Gene

Objective： To evaluate the inhibition of proliferation of human osteosarcoma cells transfected with Pin1 anti-sense gene. Methods： Different doses of antisense Pin1 gene （0, 20, 50, 100, 200, 250 μL） were transfected into osteosarcoma MG-63 cells. The cells and culture supernatant before and after transfection were collected. The curve of cell growth was made by MTT method. The cell growth cycle and apoptosis were detected by FCM. The expression of Pin1 was detected by Western-blot and that of Pin1 mRNA by polymerase chain reaction （RT-PCR） respectively. Results： MTT and FCM assays indicated that the transfection by antisense Pin1 gene could inhibit MG-63 proliferation and induce apoptosis. Western-blot assays revealed that the antisense Pin1 gene-transfected MG-63 cells had weaker staining than those without transfected with antisense Pin1 gene, and staining intensity was negatively related with doses. The cells transfected by different doses of gene （0, 20, 50, 100, 200, 250 μL） had different absorbance rate： 0.854±0.136, 0.866±0.138, 0.732±0.154, 0.611±0.121, 0.547±0.109, 0.398±0.113, 0.320±0.151 respectively, with the difference being significant by F and q test （P〈0.05）. The expression of Pin1 mRNA had the similar results and its absorbance rate was 0.983±0.125, 0.988±0.127, 0.915±0.157, 0.786±0.125, 0.608±0.124, 0.433±0.130, 0.410±0.158 respectively （P〈0.05）. Conclusion： The expression of Pin1 mRNA in MG-63 cells could be inhibited by antisense Pin1 gene, so to reduce the expression of Pin1 and depress the proliferation of human osteosarcoma cells MG-63.

Inhibitory effects of PIN1 antisense gene on the proliferation of human osteosarcoma cells

Objective： To evaluate the inhibitory effects of PIN1 antiseuse gene on the proliferation of htnnan osteosarcoma cells. Methods： Different doses of antisense PIN1 gene （0,20,50,100,200,250μl） were transfected into osteosarcoma MG-63 cells. The cells and the culture supernatants before and after transfection were collected. The cell growth curve was made using MTT method. The cell growth cycle and apoptosis were detected by FCM. The expression of PIN1 was detected by Western blot. The expression of PIN1 mRNA was detected by reverse transcription polymerase chain reaction （RT-PCR）. Results： MTT and FCM assays indicated that the transfection of antisense PIN1 gene could inhibit profiferation of MG-63 cells and lead to cell apoptosis. Western-blot assays revealed the MG-63 cells transfected with antisense PIN1 gene had weaker expression than those without transfection with antisense PIN1 gene, and the band intensity was negatively related with doses. The cells transfected with different doses of gene （0,20,50,100,200,250μl） had different absobance rate（0.854±0.136,0.866±0.138,0.732±0.154,0.611 ± 0.121, 0. 547 ± 0. 109, 0. 398 ± 0.113, 0. 320 ± 0.151）, with significant difference assessed by F and q test （P〈0.05）. The absorbance rate of PINI mRNA was 0.983±0.125,0.988±0.127, 0.915±0.157, 0.786 ± 0.125,0.608 ± 0.124,0.433 ± 0.130,0.410 ± 0. 158 respectively （ P 〈 0.05）. Conclusion. The expression of PIN1 mRNA in MG-63 cells could be inhibited by antiseuse PIN1 gene, and then the expression of PIN1 was reduced and depressed, and so the proliferation of hmnan osteosarcoma cells MG-63 was inhibited.