鳖甲煎丸通过RhoAROCK信号通路抑制肝癌细胞血管形成拟.PDFVIP

J South Med Univ, 2018, 38(8): 997-1001 doi 10.3969/j.issn. 1673-4254.2018.08.16 ·997 · 鳖甲煎丸通过鳖甲煎丸通过RhoA/ROCKRhoA/ROCK 信号通路抑制肝癌细胞血管形成拟信号通路抑制肝癌细胞血管形成拟 态的生态的生成成 安海燕，林俊豪，孙海涛，许梨梨，苏嘉琪，何春雨，曾嘉敏，梁佩湘，贺松其 南方医科大学中医药学院，广东 广州 510515 摘要：目的观察鳖甲煎丸对肝癌细胞血管生成拟态（VM）形成的作用及其对RhoA/ROCK 通路信号分子和VE-cadherin、PI3K 表达的影响，探讨鳖甲煎丸抑制肝细胞肝癌（HCC）转移侵袭的分子机制。方法 将40 只雄性SD大鼠随机分为4 组，分别以鳖甲 煎丸高（H）、中（M）、低剂量（L）及生理盐水（N ）灌胃，4 d 后采血，制备药物血清。肝癌HepG2 细胞体外Matrigel 三维培养，药物 血清及RhoA/ROCK 抑制剂Y-27632（P）干预24 h 后，应用图像采集和分析系统检测各组VM 形成情况，采用Western blotting 技 术检测各组细胞RhoA 和ROCK 1 的表达水平，ELISA 法检测各组细胞培养上清中VE-cadherin、PI3K 的含量。结果 鳖甲煎丸 可显著抑制HepG2 细胞VM 的生成，且VM 管径显著高于阴性对照组（P0.01），Y-27632 完全抑制了HepG2 细胞VM 的生成 （P0.01）；同时，鳖甲煎丸和Y-27632 都能够抑制HepG2 细胞中RhoA、ROCK 1 的表达（P0.05），降低细胞培养上清中VE- cadherin、PI3K 的表达水平（P0.05）。结论 鳖甲煎丸可抑制肝癌细胞VM 的形成，其作用机制可能与其能抑制三维培养的 HepG2 细胞中RhoA/ROCK 通路信号分子及VE-cadherin、PI3K 的表达有关。 关键词：血管生成拟态；肝癌细胞；鳖甲煎丸；Rho/ROCK ；PI3K Biejiajian Pills inhibits hepatoma carcinoma cell vasculogenic mimicry by suppressing RhoA/ROCK signaling pathway AN Haiyan, LIN Junhao, SUN Haitao, X U Lili, SU Jiaqi, HE Chunyu, ZENG Jiamin, LIANG Peixiang, HE Songqi College of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China Abstract: Objective To observe effects of Biejiajian Pills on hepatocarcinoma (HCC) cell vasculogenic mimicry (VM) and explore the molecular mechanism by which Biejiajian Pills inhibits HCC metastasis and invasion. Methods Forty male SD rats were randomly divided into 4 groups for gastric lavage of normal saline or high, moderate or low doses of Biejiajian Pills (twice daily) for 4 consecutive days. The sera were collected from the rats for treatment of cultured human HCC HepG2 cells. VM formation in the cells was detected using an image acquisition and analysis system 24 h after incubation of the cells with the sera and with the RhoA/ROCK inhibitor