背景:肝细胞癌(HCC)是一种常见的致命癌症,预后不良。Obovatol(Ob),一种新颖的木兰木兰的叶和茎皮衍生的木脂素,对多种肿瘤均有抗肿瘤作用。然而,其对肝癌的作用及机制有待进一步探讨。
方法:Huh7和Hep3B细胞,以及BALB/c裸鼠用于确定Ob对生长的功能和机制,通过细胞计数试剂盒-8,transwell,酶联免疫吸附测定(ELISA)和蛋白质印迹实验。
结果:Ob降低了Huh7和Hep3B细胞的细胞活力,IC50值为57.41µM和62.86µM,分别。Ob下降了入侵能力,N-cadherin的蛋白表达和IL-10和TGF-β的浓度,而增加了Hep3B和Huh7细胞中E-cadherin的表达以及IFN-γ和IL-2的含量。机械上,Ob降低了p-JAK/JAK的蛋白质水平,p-STAT3/STAT3和PD-L1,随着JAK/STAT3轴激活剂RO8191的治疗而部分恢复。Ob对细胞活力的影响,入侵能力,N-cadherin和E-cadherin的蛋白质水平,以及IL-10,TGF-β,在RO8191的处理下,Hep3B和Huh7细胞中的IFN-γ和IL-2被逆转。在体内,Ob减少肿瘤体积和重量,N-钙黏着蛋白的水平,PD-L1,p-JAK/JAK,和p-STAT3/STAT3,E-钙粘蛋白和IFN-γ的表达升高。
结论:Ob下调JAK/STST3/PD-L1途径以减弱生长,肝癌的侵袭和免疫逃逸。
BACKGROUND: Hepatocellular carcinoma (HCC) is a common cancer that is fatal and has a dismal prognosis. Obovatol (Ob), a novel lignan derived from the leaf and stem bark of Magnolia obovata Thunb, has exhibited anti-tumor effect on diverse tumors. However, its effect and mechanisms on HCC remain to be further explored.
METHODS: Huh7 and Hep3B cells, as well as BALB/c nude mice were used to determine the function and mechanisms of Ob on growth, invasion and immune escape by cell counting kit-8, transwell, enzyme-linked immunosorbent assay (ELISA) and western blot experiments.
RESULTS: Ob reduced the cell viability of Huh7 and Hep3B cells, with a IC50 value of 57.41 µM and 62.86 µM, respectively. Ob declined the invasion ability, the protein expression of N-cadherin and the concentrations of IL-10 and TGF-β, whereas increased the E-cadherin expression and the contents of IFN-γ and IL-2 in Hep3B and Huh7 cells. Mechanically, Ob decreased the protein level of p-JAK/JAK, p-STAT3/STAT3 and PD-L1, which was partly restored with the treatment of RO8191, an activator of JAK/STAT3 axis. The effect of Ob on the cell viability, the invasion ability, the protein level of N-cadherin and E-cadherin, and the concentrations of IL-10, TGF-β, IFN-γ and IL-2 in both Hep3B and Huh7 cells was reversed with the management of RO8191. In vivo, Ob reduced tumor volume and weight, the level of N-cadherin, PD-L1, p-JAK/JAK, and p-STAT3/STAT3, with an elevated expression of E-cadherin and IFN-γ.
CONCLUSIONS: Ob downregulated the JAK/STST3/PD-L1 pathway to attenuate the growth, invasion and immune escape of HCC.