ABSTRACT
Objective:To establish the RPB5-mediating protein (RMP)-silenced stable cell lines and study the inhibitory effects of small interfering RNA (siRNA) targeting RMP gene on the proliferation and migration of human hepatoma SMMC-7721 cells. Methods:Three RMPi siRNAs were designed and synthesized in vitro and transfected into SMMC-7721 cells. The inhibitory effect of siRNA on RMP gene expression was measured by RT-PCR to select the best siRNA. The expression vector pGPU6-Neo-RMP-484 was transfected into SMMC-7721 cells by the lipofectamine and the cells stably expressing the siRNA were selected by G418. RT-PCR was used to detect the interference efficacy against RMP gene. Cell proliferation and adhesion were measured by MTT assay. Wound healing test was used to observe the migration ability of cells. Results:The SMMC-7721 cell lines with down-regulated RMP expression were established by using RNA interference technology. Compared with the negative control cells, expression of RMP mRNA was down-regulated by(83.67±2.56)% .The proliferation of stable-transfected cells was inhibited by(74.33±0.58)% . The adhesion capability of stable-transfected cells was enhanced but the migration capacity was decreased compared with the negative control cells. Conclusion:The pGPU6-Neo-RMP-484 cell lines with stable transfection of RMP siRNA recombinant vector are successfully screened,which can be used as a cellular model for studying the molecular mechanism of RMP. Down-regulation of RMP gene expression can effectively inhibit the proliferation, enhance the adhesion, and decrease the migration of SMMC-7721 cells.
ABSTRACT
BACKGROUND: Autogenous bone has been used in cervical vertebra graft bone fusion in earliest stage and at most. However, its source is limited, simultaneously, induced many complications such as infection, hemorrhage and postoperative pain in the donor bone region. Recently, above-mentioned complications were avoided or reduced with the usage of new graft bone fusion material. OBJECTIVE: To compare clinical efficacy using MC+~R combination of autogenous bone or calcium sulfate artificial bone in antador cervical fusion.METHODS: A total of 26 patients (34 levels) with cervical spondylotic myelopathy underwent anterior cervical discectomy and cervical intervertebral fusion from January to December 2008. Anterior cervical oblique cut was 3.0-4.0 cm. The endplate were preserved after the cervical intervertebral disc and the posterior longitudinal ligament were removed. Autogenous bone group was filled with autogenous bone. Calcium sulfate artificial bone group was filled with Wdght's Osteoset artificial bone. Anchoring clip was implanted between the cervical vertebrae. Every patient had a short neck incision was assessed with X-ray, JOA grade and Odom's evaluation scale.RESULTS AND CONCLUSION: The two groups of 26 patients (34 segments)were followed up. The JOA score of postoperation was no significant difference between the two groups. According to the Odom's evaluation scale, the excellent and good rate of calcium sulfate group was higher than autogenous bone group, but there was not statistical significance (P>0.05). The fusion rate of autogenous bone group was higher than calcium sulfate group at 3 and 6 months, but the fusion rate of two groups were 100% at 12 months. Although the calcium sulfate group at 6 months, lordosis angle lost more than 0.4°than the autogenous bone group,but no significant statistically between the two groups (P>0.05). MC+ combination of autogenous bone or Calcium sulfate had the same clinical efficacy in the treatment of cervical spondylotic myelopathy, but the calcium sulfate artificial bone could be effectively avoided the complications of donor site.