小RNA干扰沉默S100A13对甲状腺细胞侵袭、迁移的影响及机制

 目的 探究干扰钙结合蛋白A13(calcium binding protein A13, S100A13)基因表达对甲状腺癌TPC-1细胞侵袭、迁移的影响及可能作用机制。方法 采用小RNA干扰敲减甲状腺癌TPC-1细胞S100A13基因的表达,通过细胞划痕实验和Transwell迁移实验检测S100A13对TPC-1细胞迁移能力的影响,Transwell侵袭实验检测S100A13对TPC-1细胞侵袭能力的影响,免疫印迹试验(Western Blot)检测S100A13、基质金属蛋白酶2(MMP-2)、钙黏蛋白E(E-cadherin)、波形蛋白(Vimentin)蛋白的表达。结果 与对照组相比,采用小RNA干扰的人甲状腺癌TPC-1细胞划痕宽度显著升高(25.214±2.687 vs 69.223±5.291),侵袭(0.897±0.082 vs 0.521±0.053)、迁移(1.263±0.252 vs 0.759±0.062)能力显著降低(P<0.05),细胞中MMP-2(0.523±0.062 vs 0.246±0.033)、Vimentin(0.863±0.082 vs 0.458±0.053)蛋白表达显著降低(P<0.05),E-cadherin(0.445±0.053 vs 0.755±0.078)蛋白表达显著增加(P<0.05)。结论 采用小RNA干扰S100A13表达可抑制甲状腺癌TPC-1细胞的侵袭、迁移,可能与其下调上皮间质转化和MMP-2的表达有关。

Effect and mechanism of sliencing S100A4 by small RNA interference of S100A13 on thyroid cells

Objective To investigate the effect of interference with the expression of calcium binding protein A13 (S100A13) on the invasion and migration of thyroid carcinoma TPC-1 cells and the possible mechanism.Methods The expression of S100A13 gene in thyroid cancer TPC-1 cells was knocked down by siRNA. The effect of S100A13 on the migration of TPC-1 cells was evaluated with the wound healing scratch test and Transwell assay. The effect of S100A13 on the invasiveness of TPC-1 cells was assessed by Transwell invasion test. The expressions of S100A13, MMP-2, E-cadherin and vimentin were detected by Western Blot.Results Compared with the control group, the silencing of S100A13 by siRNA in human thyroid carcinoma TPC-1 cells significantly increased the scratch width(25.214±2.687 vs 69.223±5.291), but the ability of invasion(0.897±0.082 vs 0.521±0.053) and migration(1.263±0.252 vs 0.759±0.062)decreased significantly (P<0.05). The levels of MMP-2 (0.523±0.062 vs 0.246±0.033)and vimentin(0.863±0.082 vs 0.458±0.053) in S100A13-treated cells were significantly decreased (P<0.05), while E-cadherin protein(0.445±0.053 vs 0.755±0.078)was significantly increased (P<0.05).Conclusions Interference with S100A13 expressions by siRNA inhibits the invasion and migration of thyroid carcinoma TPC-1 cells, which may be related to the inhibition of EMT and MMP-2.