RESUMEN
<p><b>OBJECTIVE</b>To construct a recombinant lentiviral vector that stably express hepatocyte nuclear factor 6 (HNF6) in colorectal cancer cell line and examine its effects on the invasive ability of SW620 cells.</p><p><b>METHODS</b>The lentiviral vector pLeno-DCE-HA-HNF6 was constructed and transfected into 293T cells. The supernatant containing the lentivirus particles was harvested to determine the virus titer. A stable cell line was established by infecting SW620 cells with the lentivirus particles, and the transfection efficiency was examined by fluorescence microscopy and flow cytometry. The invasion ability of the transfected SW620-HNF6 cells was assessed by wound healing and transwell assays.</p><p><b>RESULTS</b>The recombinant lentiviral vector was correctly constructed and verified by sequencing. SW620-HNF6 cell line with stable HNF6 expression was established successfully, and the transfection efficiency reached 82.3%. Western blotting and quantitative PCR demonstrated significantly upregulated HNF6 expression in SW620-HNF6 cells, which showed obviously suppressed invasive ability in wound healing and transwell assays.</p><p><b>CONCLUSION</b>We have successfully established a colorectal cancer cell line SW620-HNF6 stably expressing HNF6, which shows a lowered migration activity in vitro.</p>
Asunto(s)
Humanos , Línea Celular Tumoral , Movimiento Celular , Neoplasias Colorrectales , Metabolismo , Patología , Vectores Genéticos , Genética , Factor Nuclear 6 del Hepatocito , Genética , Lentivirus , Genética , Metabolismo , Invasividad Neoplásica , Metástasis de la Neoplasia , Proteínas Recombinantes , Genética , TransfecciónRESUMEN
Objective To evaluate the efficacy,indications and clinical outcomes of the treatment of thoracolumbar fractures by short segmental pedicle screws fixation at the level of the fracture.Methods Thirtytwo patients with thoracolumbar fracture,who underwent surgical procedure of short segmental pedicle screws fixation at the level of the fracture from 2007 to 2010,were followed up.X rays were performed preoperatively and postoperatively to locate the injured vertebral segment height and fractured kyphosis vertebral (Cobb angle).Frankel standard was used to assess the spinal cord function.Results All patients were followed up for 12 to 20 months and were in satisfying condition in the reduction of fracture.After surgery,the height of fractured vertebral body leading edge recovered from preoperative (32.4% ~69.3%,averaged (51.6 ± 17.8)% ) to (85.6% ~99.2%,averaged (92.8 ±6.2)% ) after two weeks and (90.6% ~97.8%,averaged (93.8 ±3.6)% ) at the last follow-up.Fractured vertebral Cobb angle was recovered from the preoperative ( 12.8 ° ~ 30.5 °,averaged [20.8±9.1] °) to (0° ~7.8 °,averaged [4.9 ±3.2] °) two weeks later and (2.0° ~ 12.0°,averaged [ 6.2 ± 4.6 ] o at the last follow-up.Cobb angle of the injured vertebral segment and the extend of vertebral compression were significantly improved after the angle was corrected ( P <0.01 ).Conclusion Using reduction and short segment pedicle screw fixation at the fracture level would be helpful to correct kyphotic vertebral compression and restore the height of injured vertebrate,which was also of benefit to increase the stability of short-segment posterior fixation system and reduce the loss of correction in a long run.