鸡屎藤苷酸对肝癌HepG2细胞炎症因子水平、生长、凋亡、侵袭和血管生成的影响

Effect of paederosidic acid on inflammatory factor levels, growth, apoptosis, invasion and angiogenesis of hepatocellular carcinoma HepG2 cells

关键词:  鸡屎藤苷酸  肝癌  HepG2细胞  侵袭  血管生成  VEGF

Key words:  paederosidic acid  liver cancer  HepG2 cells  invasion  angiogenesis  VEGF

摘要:

[目的] 探讨鸡屎藤苷酸对肝癌细胞系炎症因子水平、生长、凋亡、侵袭和血管生成的影响。[方法] CCK8法检测肝癌细胞系的活力；流式检测细胞凋亡；ELISA检测相关炎症因子的含量；Western blot检测B细胞淋巴2（Bcl2）、Bcl2相关X（Bax）、含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase3）、白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、E-钙黏蛋白（E-cad）、波形蛋白（Vimentin）、纤维粘连蛋白（FN）、血管内皮生长因子（VEGF）；Transwell检测细胞侵袭；内皮细胞成管实验检测肿瘤小管的形成。[结果] 当鸡屎藤苷酸溶液的浓度小于20 μg/mL时，对肝癌细胞系活力的抑制作用不具有明显差异；48 h当浓度大于40 μg/mL时，对HepG2抑制作用明显（P<0.05），且呈剂量依赖性抑制；选择HepG2细胞处理48h，鸡屎藤苷酸分成3个剂量组（0、20、40 μg/mL），厄洛替尼（0.86 ng/mL）为阳性对照。与空白对照组相比，40 μg/mL组的细胞凋亡率、抗炎因子水平（IL-10）、Bax/Bcl-2、Cleaved Caspase-3/Caspase-3和E-cad蛋白表达显著升高（P<0.05）；促炎因子水平（IFN-γ、IL-6）、侵袭细胞数量及相关蛋白表达（FN）、小管形成数量及血管内皮生长因子表达显著降低（P<0.05）。[结论] 鸡屎藤苷酸通过抗炎、抑制肝癌细胞HepG2生长、侵袭、肿瘤血管形成并促进其凋亡，缓解肝癌的发生发展。

Abstract:

[Objective] To investigate the effect of paederoside acid on the levels of inflammatory factors, growth, apoptosis, invasion and angiogenesis of hepatocarcinoma cell lines.[Methods] The viability of liver cancer cell lines were tested with CCK-8 method; Flow cytometry was applied to detect cell apoptosis; ELISA was used to detect the content of related inflammatory factors; Western blot was applied to detect Bax, Bcl-2, Caspase-3, IL-6, IL-10, E-cad, Vimentin, FN, VEGF; Cell invasion was detected with Transwell; Endothelial cell tube formation test was applied to test the formation of tumor tubules.[Results] When the concentration of paederoside acid was less than 20 μg/mL, there was no significant difference in the inhibitory effect on the viability of liver cancer cell lines; When the concentration was greater than 40 μg/mL at 48 h, the inhibitory effect on HepG2 was obvious (P<0.05), and the effect showed a dose-dependent inhibition; HepG2 cells were selected for 48h treatment in later experiments, and the experiment was divided into three dose groups (0, 20, 40 μg/mL), and erlotinib (0.86 ng/mL) was used as the positive control. Compared with the blank control group, the apoptosis rate, the level of anti-inflammatory factors (IL-10), and the expression of Bax/Bcl-2, Cleaved Caspase-3/Caspase-3 and E-cad protein in the 40 μg/mL group were significantly increased (P<0.05);The level of pro-inflammatory factors (IFN-γ, IL-6), the number of invasive cells and the expression of related proteins(Vimentin, FN), the number of tubules formed and the expression of vascular endothelial growth factor were significantly reduced(P<0.05).[Conclusion] Paederosidic acid regulates the level of inflammatory factors, inhibits the growth, invasion, tumor angiogenesis and promotes apoptosis of liver cancer cells HepG2, and alleviates the occurrence and development of liver cancer.