The Effects of Over-Expression of Fructose-1,6-Bisphosphatase 1 Gene on Reversing Malignant Biological Behavior and Inducing Osteogenic Differentiation of Osteosarcoma Cells

XIE Shengnan¹, KANG Quan² , ZHANG Yao¹, CHEN Jie¹, LUO Qing^1*

(¹Department of Pediatric Research Institute; Ministry of Education Key Laboratory of Child Development and Disorders; National Clinical Research Center for Child Health and Disorders (Chongqing); China International Science and Technology Cooperation base of Child development and Critical Disorders; Children’s Hospital of Chongqing Medical University; Chongqing Key Laboratory of Pediatrics,Chongqing 400014, China; ²Department of Hepatology,Children’s Hospital of Chongqing Medical University, Chongqing 400014, China)

Abstract:

The aim of the research is to investigate the effects of over-expressed FBP1 (fructose-1,6-bisphosphatase 1) gene on proliferation, apoptosis and migration of human osteosarcoma cell line (MG63). After the recombinant plasmid carrying FBP1 gene were transfected by liposome into MG63 cells, the expression levels of FBP1 mRNA and protein were detected by qRT-PCR and Western blot to verify the over-expression of FBP1 gene. The cell proliferation ability of MG63 cells was detected by CCK-8 assay, Trypan blue counting method and crystal violet staining. The apoptosis rates was detected by FCM and DAPI staining. Then the apotosis-associated proteins Bcl-2 and Bax were detected by Western blot. Wound-Healing asssay and transwell assay were used to detect the migration. The cell cycle was detected by Flow Cytometry. ALP staining and Alizarin red staining were used to detect the osteogenic differentiation. This study indicates that over-expressed FBP1 gene can suppress the proliferation and migration of osteosarcoma MG63 cells and enhance apoptosis significantly, in the meanwhile,early and late osteogenic differentiation were enhanced. And the proliferation was not regulated by the cell cycle.

CSTR: 32200.14.cjcb.2019.12.0003