巴戟天对人源结肠癌细胞HCT-116移植瘤的抑制作用及机制初步探讨

李灿涛; 卢颖裕; 陈勇儿; 梁健; 侯少贞; 陈鑑强

doi:10.13386/j.issn1002-0306.2021050055

巴戟天对人源结肠癌细胞HCT-116移植瘤的抑制作用及机制初步探讨

Study on the Therapeutic Effect of Morinda officinalis on HCT-116 Xenograft Tumor Model and the Underlying Mechanism

摘要

摘要: 目的：检测巴戟天对人源结肠癌细胞HCT-116移植瘤的抑制作用及初步探讨其作用机制。方法：于雄性裸鼠腋下注射人源结肠癌HCT-116细胞，建立移植瘤模型，随后采用巴戟天乙醇提取物（Ethanol extract of Morinda officinalis, EEMO）（12 mg/kg/d）和巴戟天生药（Crude drug of Morinda officinalis, CDMO）（200 mg/kg/d）为实验组进行灌胃给药，以5-氟尿嘧啶（5-Fluorouracil, 5-FU）为阳性药（30 mg/kg，每3 d给药一次，腹腔注射），以水（一次/d）为对照组，共四组（EEMO组、CDMO组、5-FU组和对照组），持续给药23 d。通过检测各干预组动物的肿瘤重量、肿瘤体积、肿瘤组织病理变化判断巴戟天的抑癌作用；通过检测肿瘤组织血管生成因子（VEGF）、缺氧诱导因子-1α（HIF-1α）和环氧化酶-2（COX-2）的表达水平以及巨噬细胞的极化趋势初步探讨其抑癌途径。结果：EEMO和CDMO能不同程度抑制小鼠移植瘤体积的增长（P<0.05）、降低移植瘤的重量（P<0.01）和降低血清前列腺素E2（PGE2）的水平（P<0.05）。蛋白免疫印迹和免疫组化的检测结果显示EEMO和CDMO可显著下调肿瘤组织VEGF、HIF-1α和COX-2的表达水平（P<0.01和P<0.05）。免疫荧光检测结果显示，EEMO和CDMO极显著增加iNOS的表达水平（P<0.01），并且极显著提高iNOS/CD206的比例（P<0.01）。结论：EEMO和CDMO对小鼠体内的移植瘤发生有抑制作用，可能与其下调VEGF、HIF-1α和COX-2/PGE2信号通路的表达、增强巨噬细胞向M1型极化有关。

Abstract: Objective: To explore the inhibitive effect and underlying mechanism of Morinda officinalis (MO) on human colorectal cancer cell HCT-116 xenograft tumor model. Methods: Xenograft tumor model was established through transplanting human colorectal cancer cell into the armpits of male nude mice. Four groups were divided for appropriate administration, including ethanol extract of Morinda officinalis (EEMO) group (12 mg/kg/d), crude drug of Morinda officinalis (CDMO) group(200 mg/kg/d), 5-fluorouracil (30 mg/kg, once per three days) group, control group (water, once a day). The administration of drugs was lasted for 23 days. Tumor weight, tumor volume and pathological changes were detected. Protein expression of Hypoxia inducible factor-1α (HIF-1α), Cyclooxygenase-2/Prostaglandin E2 (COX-2/PGE2) pathway and macrophage polarization were determined after the experiment. Results: Both EEMO and CDMO could significantly inhibited the weight and growth of HCT-116 xenograft tumor (P<0.01), lowered the content of PGE2 (P<0.05), reduced the expression of HIF-1α, COX-2 and VEGF in tumor (P<0.01 and P<0.05). Additionally, EEMO and CDMO could remarkably increase the ratio of M1-like macrophage phenotype in tumor microenvironment (P<0.01). Conclusion: MO exerted anti-tumor effect partly via inhibiting expression of HIF-1α, COX-2/PGE2 pathway and polarizing tumor-associated macrophages toward M1-like macrophage phenotype.