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ROCK1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(P<0.01),Y-27632完全抑制了HepG2细胞VM的生成(P<0.01);同时,鳖甲煎丸和Y-27632都能够抑制HepG2细胞中RhoA、ROCK1的表达(P<0.05),降低细胞培养上清中VEcadherin、PI3K的表达水平(P<0.05)。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Hep G2细胞中RhoA/ROCK通路信号分子及VE-cadherin、PI3K的表达有关。
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 Y-27632（P）. The expression levels of RhoA and ROCK1 in the cells were detected using Western blotting, and the contents of VE-cadherin and PI3 K in the culture supernatant were determined using ELISA.Results Treatment with the sera from Biejiajian Pills-treated rats significantly inhibited formation of VM in HepG2 cells, and the diameters of VM formed were significantly greater than those in the positive control group（P〈0.01）. Y-27632 completely inhibited the formation of VM in HepG2 cells（P〈0.01）. Treatments with Biejiajian Pills and Y-27632 both inhibited the expression of RhoA and ROCK1（P〈0.05） and significantly lowered the contents of VE-cadherin and PI3 K in the culture supernatant（P〈0.05）. Conclusion Biejiajian Pills can inhibit the formation of VM in HCC cells in vitro possibly by inhibiting the RhoA/ROCK pathways and the expressions of VE-cadherin and PI3 K.
作者
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
AN Haiyan;LIN Junhao;SUN Haitao;XU Lili;SU Jiaqi;HE Chunyu;ZENG Jiamin;LIANG Peixiang;HE Songqi(College of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2018年第8期997-1001,共5页
Journal of Southern Medical University
基金
国家自然科学基金(81603427)
广东省自然科学基金(2016A030313596)~~
