摘要目的 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。
Abstract: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.
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