The Mechanism Preliminary Study For Serum-deprivation -induced Release Of S100A13 And FGF-1 From Thyroid Cancer TT Cell

Objective: To investigate the effect of serum-deprivation-induced on the changes of [Ca2+]i and the release of S100A13 and FGF1 in thyroid cancer TT cell.Then, to preliminary reveal the role of Ca2+ on the release of S100A13 and FGF-1.Methods:The protein expression quantity of FGF-1 and S100A13 in TT cells on time 0h,1h,2h,4h,6h under serum-deprivation were detected by Western Blot,The release of FGF-1from TT cells on time0h,1h,2h,4h,6h under serum-deprivation were detected by ELISA.Real-time dynamic examines on the change of 1h[Ca2+]i in TT cells under serum-deprivation were detected by confocal laser scanning microscopy.Then, The effect of EGTA(2.5mmol/L),BAPTA-AM(2.5μmol/L)on distributions of the florescence of S100A13 and FGF-1 in TT cells under serum-deprivation 6h were detected by indirect immunofluorescence, Such effect on the release of both proteins were detected by Western-Blot and ELISA.Results: Western Blot showed that the expression of FGF-1 and S100A13 on time 0h,1h,2h,4h,6h in cells under serum-deprivation Has gradually the declining trend. 0h, 1h, the 2h group compares not obvious difference, 4h group compares With 0h, 1h, 2h has statistics significance(P<0.05),6h group cell FGF-1,S100A13 protein expression obvious drop, With 0h, 1h, 2h, the 4h group difference has statistics significance(P<0.01).ELISA showed FGF-1 on time 0h, 1h, 2h, 4h, 6h group cell culture clear FGF-1 the density has the rise tendency.the 4h group raises the clear FGF-1 density to compare 0h, 1h, and 2h group to elevate obviously(P<0.05).the 6h group raises the clear FGF-1 density to compare 0h, 1h, 2h, and 4h group to elevate obviously(P<0.01).Confocal laser scanning of Ca2+ imaging indicated that [Ca2+]i of normal group cells were stable respectively all the scanning times, about 0.1～0.3umol/l.[Ca2+]i of serum-deprivation cells were also stable within 23 min, but accelerated to 1.6umol/l, Maintains about 17min,after 40min slow drop to a low level.From 40min to 60min[Ca2+]i close 0.3~0.6μmol/l. the average [Ca2+]i was higher than normal group (p<0.05). Both of EGTA and BAPTA-AM could inhibit the rise of [Ca2+]i by serum-deprivation,the average [Ca2+]i was similar with normal group.Indirect immunofluorescence indicated the florescence intensitys of both S100A13 and FGF-1 in cells under serum-deprivation 6h were weakened; EGTA,BAPTA-AM could reverse the changes. Both expression of S100A13 and FGF-1 in cells under serum-deprivation 6h were lower than normal group, indicated by Western Blot. But addition of EGTA or BAPTA-AM could make both expression of the two proteins as the same as normal group.ELISA showed that FGF-1 of serum-deprivation 6h group in cell medium was higher than other 3 groups,obviously (P<0.01).Conlusion:Ca2+ is possible related with the release of S100A13 and FGF-1 in TT cell under serum-deprivation-induced;Both the Ca2+-Chelating agent EGTA and BAPTA-AM could inhibit the rise of [Ca2+]i in cells and the release of S100A13 and FGF-1 by Serum-deprivation-induced.