The Effect Of S100A13 Gene On The Growth In Human Thyroid Cancer Cell Line TT

OBJECTIVE: This study was to construct TT cells of overexpressing S100A13 gene stably and to investigate the effect of exogenous S100A13 overexpression on the proliferation of human thyroid cancer cell line TT.METHODS: The eukaryotic expression plasmid pcDNA3.2/ V5/GW/D-S100A13 and empty vector pcDNA3.2/ V5/GW/D were transfected into TT cells. The cells were selected by G418. The expression of S100A13 mRNA and protein was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The cell growth after transfection was detected by phase contrast microscope .The effects of S100A13 on cell growth proliferation and cell cycle progression were measured by cell growth curve,MTT colorimetric method and flow cytometry.RESULTS: TT-S100A13-V5 and TT-V5 cells, which separately expressed S100A13 and pcDNA3.2/ V5/GW/D vector, were constructed successfully. TT-V5-S100A13 cells grew faster than TT-V5 and TT cells[The cell count was(2.01±0.05)×105 vs.(0.89±0.04)×105 and(0.81±0.03)×105 at the 7th day of cell culture.P<0.001;The relative growth rate was (92.11±0.51)% vs.(44.09±0.75)% and(45.01±0.65)% at the 9th day of cell culture. P<0.001],which was detected by phase contrast microscope after transfection;Both S phase proportion and G2/M phase proportion were significantly higher in TT-V5-S100A13 cells than in TT-V5 and TT cells [(9.12±0.32)% vs. (6.22±0.12)% and (6.18±0.31)% at S phase, P<0.001; (26.95±0.58)% vs. (11.33±0.39)% and (12.01±0.26)% at G2/M phase, P<0.001].CONLUSION: The cell line of overexpression exogenous S100A13 gene which taped with V5 small fragments of pcDNA3.2 vector was constructed successfully . Overexpression exogenous S100A13 gene could accelerate cell growth, and promote cell cycle progression of TT cells from G0/G1 phase to S and G2/M phase.