扶正抑癌方含药血清对结肠癌lovo细胞E-cad、N-cad和Snail蛋白表达的影响

Effect of Fuzheng Yi'ai Decoction-containing Serum on the Expressions of E-cad, N-cad and Snail Proteins in Colon Carcinoma Cell Line Lovo

[目的]通过观察扶正抑癌方含药血清对结肠癌lovo细胞E-cad、N-cad和Snail蛋白表达的影响,探讨扶正抑癌方对结肠癌细胞EMT的影响,从而进一步明确扶正抑癌方抑制结肠癌转移的机制。[方法]lovo细胞体外培养,制备大鼠不同浓度含药血清后,MTT法测定不同浓度含药血清对细胞增殖的影响;免疫荧光法检测E-cad、N-cad的表达,Western Blot检测Snail蛋白的表达。[结果]高剂量含药血清组24h对lovo细胞抑制率较中剂量、低剂量组更加明显,差异具有统计学意义(P<0.01),免疫荧光结果显示扶正抑癌方含药血清可以下调lovo细胞N-cad的表达和上调E-cad表达,与正常组和空白组相比,差异具有统计学意义(P<0.05);Western Blot检测结果显示扶正抑癌方含药血清能降低lovo细胞转录因子Snail蛋白的表达,与正常组和空白组比较差异具有统计学意义(P<0.05)。[结论]扶正抑癌方含药血清通过上调E-cad表达和下调N-cad的表达,抑制lovo细胞的EMT转变,从而降低lovo细胞的侵袭力和运动能力。

[Purpose] To investigate the effect of Fuzheng Yi＇ai Decoction-containing serum on the expressions of E-cad, N-cad, Snail proteins in colon carcinoma cell line lovo, in order to explore the inhibiting effect of Fuzheng Yi＇ai Decoction on the metastasis of colon carcinoma. [Methods] Cell line lovo was cultured in vitro. Different concentrations of Fuzheng Yi＇ai Decoction-containing serum were prepared, and their proliferation effects on cell line lovo were observed by MTT method. The expressions of E-cad and N-cad proteins were detected by immunofluorescence assay, and the expression of Snail protein was detected by Western Blot assay.[Results] The high concentration group of drug serum had a higher cell inhibition effect at 24 hours, compared with the medium group and low group （P0.01）. The expressions of N-cad and Snail proteins in drug serum group were down-regulated and the expression of E-cad protein was up-regulated, compared with the blank and normal control group （P0.05）. [Conclusion] Fuzheng Yi＇ai Decoction-containing serum can inhibit the EMT transition of cell line lovo and decrease its migration and invasive ability through the down-regulation of N-cad and Snail expressions and the up-regulation of E-cad expression.