Molecular Mechanisms Of RhoC Inducing Invasion And Metastasis Of Osteosarcoma

Objection: To research the molecular mechanism of RhoC inducing the invasion and metastasis of osteosarcoma.Methods: Culture SOSP-9607H9 and SOSP-9607E10 cell lines, and compare the reproductive activity of these two cell lines with growth curve. Detect the RhoC mRNA and protein expression levels in the two cell lines by using RT-PCR and Western-blot. Further compare the migration ability of these two cell lines by Wounding heal assay and Transwell assay, and analyze the relationship with the invasion/metastasis potency and the expression levels of RhoC. Construct the pCDNA3.1-RhoC plasmid and transfect it to SOSP-9607H9 cell line (with low invasion/metastasis potency) by Life2000. Then the over-expression cell model is made after G418 screening and the over-expression of RhoC inducing the invasion and metastasis of osteosarcoma is testified reversely. Design the siRNA fragment of RhoC and ligate it to T4 vector, and transfect the MSCVneo-SiRhoC to retrovirus incasing cells PT67 by calcium phosphate precipitation method. Then the virus supernatantis collected to silence the expression in SOSP-9607E10 cell lines which high-express RhoC, and the Wounding heal assay and Transwell assay are performed again to detect the migration ability of these two cell lines.Result:SOSP-9607 E10 cell line shows enhanced ability of proliferation in the exponential phase of growth although there is no deference between the two cell lines in the early phase after inoculation.