华蟾素对胃癌肿瘤细胞SGC-7901增殖、凋亡以及JAK/STAT3信号通路的影响

Effect of cinobufacin on the proliferation and apoptosis of human gastric carcinoma cell SGC-7901 and JAK/STAT3 signaling pathway

目的探讨华蟾素(Cino)对转录激活因子3(STAT3)的调控作用,以及对人胃癌肿瘤细胞SGC-7901的增殖、凋亡的影响。方法采用MTT法检测不同浓度华蟾素(0.1、1.0、10 mg/L)及5-FU(20 mg/L)分别作用于人胃癌肿瘤SGC-7901细胞12、24、48 h后对肿瘤细胞增殖活力的影响;半定量RT-PCR检测Cyclin D1、PCNA、Bcl-2的mRNA表达水平;蛋白悬液芯片(Bio-plex)法检测STAT3磷酸化(p-STAT3)蛋白表达水平。结果华蟾素可显著抑制SGC-7901的增殖,且呈时间和剂量依赖性。华蟾素能抑制SGC-7901细胞Cyclin D1、PCNA、Bcl-2的mRNA表达水平(均P<0.05)。华蟾素抑制p-STAT3蛋白表达水平,且呈现剂量依赖性(P<0.05)。结论华蟾素具有抑制胃癌肿瘤细胞增殖和诱导凋亡的作用,其可能机制与其抑制STAT3活化,从而抑制其下游靶基因表达有关。

Objective To explore the regulating effect of cinobufacin on signal transducer and activator of transcription 3（STAT3） and the effect on the proliferation and apoptosis in human gastric carcinoma cell SGC-7901. Methods SGC-7901 cells were incubated with the cinobufacin at different concentrations（0.1, 1.0, 10 mg/L） and 5-FU（20 mg/L） for 12, 24, and48 h, respectively. MTT method was used to measure the effect on the proliferation activity of SGC-7901 cells. Semiquantitative RT-PCR was used to determine the mRNA expression of Cyclin D1, PCNA and Bcl-2 in SGC-7901 cells. The protein expression of phosphorylated STAT3（p-STAT3） in SGC-7901 cells was detected by Bio-plex. Results Cinobufacin inhibited the proliferation of SGC-7901 cells in a dose-and time-dependent manner. Cinobufacin inhibited the mRNA expression level of Cyclin D1, PCNA and Bcl-2 of SGC-7901 cells（P〈0.05）. Moreover, cinobufacin decreased the protein expression of p-STAT3（P〈0.05） in a dose dependent manner. Conclusion Cinobufacin can inhibit the proliferation and induce apoptosis in gastric cancer cells. The possible mechanism may be related to that it inhibits activation of STAT3 and thus inhibit the expression of downstream target gene.