EGCG 抑制 Nicotine 诱导肺腺癌 H1299细胞JAK2/STAT3 mRNA 的表达研究

摘要

In order to explore the inhibitory effect of EGCG on nicotine which induced lung cancer cells proliferation, this study selected the optimal concentrations of EGCG and nicotine on lung adenocarcinoma cancer cell H1299 by MTT experiment, and detected mRNA relative expression levels of Bax, Bcl-2, Jak2 and Stat3 genes in H1299 cells using qRT-PCR technique. Experiment results showed that the IC50 of EGCG on H1299 lung adenocarcinoma cells is about 32 μmol·L-1 (treated 48 h) and 15 μmol·L-1 (treated 24 h), nicotine obviously promotes the proliferation of H1299 lung adenocarcinoma cell at 1 μmol·L-1 concentration treatment. Pretreatment of H1299 cells with 15μmol·L-1 EGCG (24 h) can significantly down-regulate the promotion of proliferation induced by 1 μmol·L-1 Nicotine (P<0.05). Using 1 μmol·L-1 nicotine to treat H1299 cells has showed that nicotine significantly reduce the mRNA expression level of Bax gene, increase the mRNA expression levels of Bcl-2, Jak2 and Stat3 genes in the JAK2/STAT3 signal pathway. Pretreatment of H1299 cells with 15 μmol·L-1 EGCG (24 h) can be observed that EGCG significantly retroregulated the mRNA expression levels of Bax, Bcl-2, Jak2 and Stat3 genes in the JAK2/STAT3 signal pathway, and got the results with significant differences (P<0.05). In conclusion, EGCG plays important role in suppressing the survival rates of H1299 cells and mRNA expression level of JAK2/STAT3 signaling pathway induced by nicotine.%为探索 EGCG 对 Nicotine 诱导肺癌细胞增殖的抑制作用，本研究通过 MTT 实验筛选出 EGCG、Nicotine对肺腺癌细胞 H1299的最佳作用浓度，利用实时荧光定量 PCR 技术检测 EGCG、Nicotine 对 H1299细胞中Bax、Bcl-2、Jak2和 Stat3基因 mRNA 相对表达量的变化。实验结果表明，EGCG 对 H1299细胞的半抑制浓度IC50值约为32μmol·L-1（24 h）、15μmol·L-1（48 h）；1μmol·L-1的 Nicotine 对 H1299细胞促增殖作用明显；以15μmol·L-1的 EGCG 预处理 H1299细胞24 h 可显著下调 1μmol·L-1 Nicotine 的促增殖作用（P<0.05）。1μmol·L-1的 Nicotine 处理 H1299细胞可明显降低 JAK2/STAT3信号通路中 Bax 基因 mRNA 的表达，增加 Bcl-2、Jak2、Stat3基因的 mRNA 表达；15μmol·L-1 EGCG 预处理 H1299细胞可反向调控 Nicotine 诱导所致JAK2/STAT3信号通路中 Jak2、Stat3和 Bax、Bcl-2基因表达量的变化，结果具有显著性差异（P<0.05）。由此可知，EGCG 对 Nicotine 诱导的肺腺癌 H1299细胞增殖及 JAK2/STAT3信号通路中促增殖基因 mRNA 的表达起抑制作用。