The Effect Of BRAF V600E On Biological Characteristics Of Human Pancreatic Cancer Cell Line Panc-1

Objective: To investigate the effect of BRAF V600E on the biological behaviors of human pancreatic cancer cell line Panc-1 by transferring BRAF V600E into Panc-1 cell, which will provide further theoretical basis for the study of pathogenesis and treatment of pancreatic cancer.Methods: Human pancreas cell lines Panc-1 were transferred with BRAF V600E through the liposome, and the cells with BRAF V600E were detected through G418 and further confirmed through the methods of polymerase chain reaction (RT-PCR) and Western blot. And Panc-1 cells transferred with BRAFW and Panc-1 cells transferred with empty carrier were taken as the negative control and blank control respectively. The cells were incubated in DMEM with 10% FBS, G418, 2mM Glutamine, 100U/mL penicillin and 100U/mL streptomycin at 37℃with 5% CO2. MTT assay: 2,000 cells of each clone were plated per well onto 96-well microtiter plates filled with DMEM containing 10% FBS and G418. On 0h, 24h, 48h and 72h, the cells were dislodged and the assay was initiated by adding 10μL MTT to each well. And after 4-hour incubation at 37℃with 5% CO2, the medium was removed and 150μL DMSO was added to each well. Plates were read at a wavelength of 570nm. Flow cytometry analysis: 1×106 cells of each clone were vaccinated in the 50mL culture bottle, and incubated with 100μg/mL propidium iodide(PI) and 10mg/mL RNase A at 37℃with 5% CO2 for at least 30min. Samples were subjected to Flow cytometry. The migration and invasion assay: (1) The migration assay: 1×105 cells...