Abstract:Objective: To screen out the Hub gene and analyze its correlation with osteoporosis by sequencing the genes of MC3T3-E1 cells under continuously compressive pressures (CCP) . Methods: MTT assay was used to detect cell proliferation and immunofluorescence assay was used to screen out the best experimental conditions. RNA was extracted by TRIzol method and sequenced. And by Mfuzz clustering, enrichment analysis, GSVA analysis, NCBI GEO database, Genecards database and molecular signatures database, the differentially expressed genes in MC3T3-E1 cells under CCP were analyzed and compared, the Hub gene was screened, and the molecular mechanism associated with osteoporosis signal pathway was compared and analyzed. Results: The proliferation of MC3T3-E1 cells decreased significantly under CCP, and tubulin and actin inhibited significantly in the cytoskeleton of cell model groups treated with 1.0 MPa for 8 hours and 0.5 MPa for 24 hours. After comparing the blank group and the model group, 29 164 different genes were found, of which 14 489 were up-regulated and 14 675 were down-regulated. These differential genes involved 1 658 GO functions, including 1 096 GO_BP (biological process), 255 GO_CC (cell component) and 307 GO_MF (molecular function), and there were 305 signal transduction pathways regulated by differential genes in MC3T3-E1 cells under CCP. The final screened Hub genes were BTK, CSF1R, MATK, NOS1 and PDGFRB. The expression of MATK in osteoporosis was significant (P<0.05) . The areas under AUC curve corresponding to the five core genes were BTK (0.900), CSF1R (0.850), MATK (0.950), NOS1 (0.650) and PDGFRB (0.800) . According to GSVA analysis of five core genes, MATK was involved in three key signaling pathways: INFLAMMATORY-RESPONSE, HEDGEHOG-SIGNALING and KRAS-SIGNALING-DN. Conclusion: CCP can decrease the proliferation activity of MC3T3-E1 cells, and reduce the influence of tubulin and actin in cytoskeleton. MATK may be a gene that regulates the osteogenesis of MC3T3-E1 cells under CCP, and is related to three key signal transmission pathways of osteoporosis.
