葡萄籽提取物原花青素诱导人食管癌细胞ECA109凋亡

目的:探讨葡萄籽提取物原花青素(grape seed proanthocyanidins extract,GSPE)诱导人食管癌细胞ECA109凋亡的效果及机制。方法:ECA109细胞用0、25、50、100、200、400μg/mL GSPE干预24 h,噻唑蓝法检测并计算不同质量浓度GSPE对ECA109细胞的活性抑制情况,选取半数抑制浓度(50μg/mL)作为干预剂量;流式细胞技术检测细胞凋亡情况;酶联免疫吸附检测法测定炎性因子及Bax/Bcl-2表达情况;实时荧光定量聚合酶链式反应(real-time polymerase chain reaction,qPCR)、Western blot检测核因子κB(nuclear factor-kappa B,NF-κB)通路和凋亡相关蛋白Caspase-3的表达情况。结果:不同质量浓度GSPE作用24 h均可抑制ECA109细胞的增殖,细胞凋亡率由54.44%增加到82.80%;酶联免疫吸附检测结果显示,相比于空白对照组,GSPE和NF-κB抑制剂BAY11-7082可显著抑制细胞内炎性因子前列腺素E_2(prostaglandin E_2,PGE_2)的产生以及C型反应蛋白(C reactive protein,CRP)的分泌(P0.05);qPCR和Western blot结果表明GSPE和BAY11-7082处理使Caspase-3 mRNA和蛋白相对表达量显著升高(P0.05),NF-κB信号通路中IκB、p65、p50 mRNA表达量显著降低(P0.05),p-IκB、p65、p50相对表达量显著下降(P0.05)。结论:GSPE可通过抑制PGE_2、CRP的产生诱导ECA109细胞凋亡,其机制可能与抑制NF-κB通路、活化Bax有关。

Effect of Grape Seed Proanthocyanidins Extract on Apoptosis Induction in Human Esophageal Cancer Cells ECA109

Purpose: Grape seed proanthocyanidins extract (GSPE) is considered to be an antitumor bioactivator. This study aims to explore the effectiveness of GSPE in inducing cell apoptosis in human esophageal cancer cell line ECA109 and to elucidate the underlying mechanism. Methods: ECA109 cells were treated with 10 μmol/L BAY11-7082 and GSPE at concentrations of 0, 25, 50, 100, 200, 400 μg/mL for 24 h. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and flow cytometry were used respectively to measure cell proliferation and apoptosis; inflammatory cytokines and Bax/Bcl-2 were detected by enzyme-linked immunosorbent assay (ELISA) with double antibodies; the mRNA and protein expression of caspase-3 and nuclear factor-kappa B (NF-κB) signaling was detected by real-time polymerase chain reaction (qPCR) and Western blot. Results: GSPE at all concentrations tested could inhibit cell proliferation and induce apoptosis in ECA109 cells, increasing the apoptotic rate from 54.44% to 82.80%. ELISA results showed that GSPE and BAY11-7082 both could significantly inhibit the secretion of prostaglandin E2 (PGE2) and C reactive protein (CRP) (P < 0.05). The results of qPCR and Western blot showed that BAY11-7082 and GSPE could activate caspase-3 and suppress NF-κB signaling. Conclusion: GSPE can induce cell apoptosis and inhibit proliferation in ECA109 cells by inhibiting the production of inflammatory factors through suppressing NF-κB signaling pathway and activating caspase-3.