Background: It has been proven that chlorogenic acids can produce anticancer effects by regulating cell cycle, inducing apoptosis, inhibiting cell growth, Notch signaling pathways are closely related to many human tumors. The aim of this study is to study the mechanism of chlorogenic acid on apoptosis of non-small lung cancer through Notch1 pathway in animal level, and hope to provide theory basis on clinical treatment and research aimed at targeting Notch1 signaling in non-small cell carcinoma (NSCLC).
Methods: MTT assay was used to evaluate the A549 cell proliferation under the treatment of chlorogenic acid. The effect of chlorogenic acid on apoptotic and cell cycle were detected by flow cytometry. The animal model of A549 cell transplanted in nude was established, tumer size and weight were detected. The mRNA level of Notch1 signal pathway related facter were detected by RT-PCR; the expression of Notch1 signal pathway related facter in tumor tissue was detected by western blot.
Results: Chlorogenic acid inhibited the A549 cell proliferation. incresed cell apoptotic and cell percentagein G2/M (P<0.05), and in a dose-dependent manner. In animal model, tumer size and weight were lower than control group, the difference was statistically significant (P<0.05). The relative expression of mRNA of Notch1, VEGF, Delta4, HES1 and HEY1 were decreaced (P<0.05) in tumor tissue which treated with chlorogenic. The expression of Notch1 were decreaced, PTEN, p-PTEN, p-AKT were increced significantly in tumor tissue which treated with chlorogenic (P<0.05).
Conclusions: Chlorogenic acid can regulate theapoptosis of non-small lung cancer through Notch pathway in animal level, which may be associated with the down-regulating the expression of VEGF and Delta4. Notch pathway may cross talk with PI3K/AKT pathway through PTEN in NSCLC.
背景与目的 绿原酸类物质可通过调节细胞周期、诱导凋亡、抑制细胞生长等途径产生抗癌作用，Notch信号通路与人类许多肿瘤都存在密切的关系，本研究旨在探讨绿原酸通过Notch1信号通路控制非小细胞肺癌细胞凋亡的作用机制，为临床应用以及Notch1靶向药物的研究提供依据。方法 MTT法检测不同浓度的绿原酸对非小细胞肺癌细胞系A549细胞形态和细胞增殖的影响；流式细胞仪检测绿原酸对A549细胞的凋亡和细胞周期的影响；建立A549细胞的裸鼠荷瘤模型；测量肿瘤大小和重量；实时荧光定量PCR法检测Notch信号通路相关因子的mRNA表达水平；免疫印迹法检测Notch信号通路相关因子的蛋白表达水平。结果 绿原酸抑制A549细胞增殖，诱导A549细胞凋亡，增加细胞G2期/M期细胞百分比增加（P<0.05），并且呈现剂量依赖趋势。在A549细胞的裸鼠荷瘤模型中，实验组肿瘤大小和体积明显小于对照组，差异具有统计学意义（P<0.01）。试验组Notch1、VEGF、Delta4、HES1、HEY1 mRNA表达量较对照组明显减少（P<0.05）。实验组Notch1蛋白明显减少，PTEN、p-PTEN、p-AKT明显增加（P<0.05）。结论 在动物水平，绿原酸可能通过Notch1信号通路调控非小细胞肺癌的凋亡，可能是通过减少VEGF的表达，下调Delta 4水平，从而抑制Notch1信号通路的活化。Notch1信号通路可能通过PTEN与PI3K/AKT通路存在交叉调控作用。.