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

• Published in: The Chinese-German journal of clinical oncology Vol. 5; no. 4; pp. 294 - 297
• Main Authors: Xiong, Wenhua, Chen, Anmin, Guo, Fengjin
• Format: Journal Article
• Language: English
• Published: Department of Orthopaedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology,Wuhan 430030, China 01.08.2006
• Subjects: 基因转染; 核苷酸; 肿瘤细胞; 骨肉瘤; Pin1; antisense nucleotide; gene transfection; osteosarcoma
• ISSN: 1610-1979; 1613-9089
• DOI: 10.1007/s10330-005-0411-8

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.