目的:研究铜螯合剂(四硫钼酸铵,TM)联合乐伐替尼治疗肝细胞癌。
方法:将55只C57小鼠右大腿后皮下注射Hepa1-6肝癌细胞悬液。7天后,皮下肿瘤形成,将小鼠随机分为五组:TM(G1),Lenvatinib(G2),TM+Lenvatinib(G3),控制(G4),和葡萄糖酸铜(II)(G5)。在预定时间点测量血清和肿瘤中的铜浓度。经过14天的治疗,分析肿瘤重量和体积,组织学观察,免疫组化法检测肿瘤组织中血管内皮生长因子(VEGF)的表达和微血管密度(MVD)。
结果:正常小鼠血清中的铜浓度中位数分别为401.70、469.40和665.35µg/L,在小鼠植入后7天,在对照组中,分别。植入后7天G4小鼠瘤内铜浓度高于小鼠(P<0.05)。G5组的血清铜浓度高于所有其他组(P<0.05;(G1,G2和G3)。G4,P<0.05;G1vs.G2,P=0.013;G2vs.G3,P=0.018;G1vs.G3,P=0.903。G1、G2、G3和G5瘤内铜浓度分别为608.40、980.00、539.31和2938.90µg/L。G1、G2、G3、G4和G5中的平均肿瘤重量分别为0.55、0.44、0.08、1.37和3.11。G5vs.其他团体,P<0.05;(G1、G2和G3)vs.G4,P<0.05;G1vs.G3,P<0.05;G2vs.G3,P<0.05;G1vs.G2,P>0.05。此外,VEGF在G1、G2和G3中的表达水平显著低于G4和G5(P<0.05)。在五组中观察到MVD的类似趋势,但在G1和G2没有检测到显著差异。
结论:研究显示肿瘤负荷与铜之间存在显著正相关。铜促进肿瘤进展,但铜螯合抑制肿瘤生长.TM与lenvatinib的组合减少了肿瘤血管生成并提高了抗肿瘤治疗的效果。这些发现是联合疗法临床应用的基础。
The study aims to investigate the suppressing tumor-promoting effects via multi-anti-angiogenesis activity of the copper chelator (ammonium tetrathiomolybdate, TM) combined with lenvatinib for hepatocellular carcinoma.
A total of 55 C57 mice were injected subcutaneously with Hepa1-6 hepatoma cell suspensions into the right posterior thigh. After 7 days, the subcutaneous tumors were formed, and the mice were randomly divided into five groups: TM (G1), Lenvatinib (G2), TM+Lenvatinib (G3), Control (G4), and Copper (II) Gluconate (G5). The copper concentrations in serum and tumors were measured at the predetermined time points. After 14 days of treatments, tumor weight and volumes were analyzed, histology was observed, and the expressions of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in tumor tissues were measured by immunohistochemistry.
The median concentration of copper in serum was 401.70, 469.40, and 665.35 μg/L in normal mice, in mice 7 days after implantation, and in the control group, respectively. The intratumoral copper concentrations were higher in G4 mice than in mice 7 days after implantation (P < 0.05). The serum concentration of copper was higher in G5 than all the other groups (P < 0.05; (G1, G2, and G3) vs. G4, P < 0.05; G1 vs. G2, P = 0.013; G2 vs. G3, P = 0.018; G1 vs. G3, P = 0.903. The intratumoral copper concentrations were 608.40, 980.00, 539.31, and 2938.90 μg/L in G1, G2, G3, and G5, respectively. The average tumor weight was 0.55, 0.44, 0.08, 1.37, and 3.11 in G1, G2, G3, G4, and G5, respectively. G5 vs. other groups, P < 0.05; (G1, G2, and G3) vs. G4, P < 0.05; G1 vs. G3, P < 0.05; G2 vs. G3, P < 0.05; G1 vs. G2, P > 0.05. Furthermore, the expression levels of VEGF were significantly lower in G1, G2, and G3 than in G4 and G5 (P < 0.05). A similar trend was observed for MVD in the five groups, but no significant difference was detected in G1 and G2.
The study showed a significant positive correlation between tumor load and copper. Copper promotes tumor progression, but copper chelating suppresses tumor growth. The combination of TM with lenvatinib reduces tumor angiogenesis and improves the effect of antitumor treatment. These findings underlie the clinical application of combination therapy.