Zhou Jing, Li Yaocheng, Shi Xuejing, Hao Shulan, Zhang Fupeng, Guo Zhi, Gao Yu, Guo Hao, Liu Likun
Department of Oncology, Shanxi Province Academy of Traditional Chinese Medicine, Shanxi Province Hospital of Traditional Chinese Medicine, Taiyuan, Shanxi 030012, China.
Department of Anesthesiology, Shanxi provincial people's Hospital, Taiyuan, Shanxi 030000, China.
J Cancer. 2021 Apr 2;12(11):3257-3264. doi: 10.7150/jca.55929. eCollection 2021.
Tumor blood vessels exhibit morphological and functional aberrancies. Its maturity and functionality are closely associated with colon cancer progression and therapeutic efficacy. The direct evidence proving whether oridonin (ORI) has vascular normalization promoting effect from which combination therapies will benefit is still lacking. We established a subcutaneous xenograft model of human colon cancer. The animals were divided into the Control and ORI-treated groups. Immunohistochemical analysis and TUNEL staining was applied to evaluate the proliferation, apoptosis and angiogenesis. Western blot analysis was employed to characterize the angiogenesis-related factors and JAK2/STAT3 signaling. Then, vascular normalization and macrophage reprogramming were assessed by immunofluorescence analysis. The results showed that ORI obviously reduced tumor growth, diminished the numbers of Ki67 cells and CD31 microvessel density, while increased the numbers of TUNEL cells. The expression levels of VEGF and bFGF proteins were dramatically down-regulated while the angiostatin and endostatin levels were increased in the ORI-treated group. Moreover, ORI therapy remarkably promoted the pericyte coverage of tumor vessels from days 5 to 10, with the highest pericyte coverage rate occurred at day 7. In the time window of vascular normalization, hypoxia of the tumor microenvironment was improved by ORI, the expression of HIF-1a was downregulated. Moreover, CD206 macrophage cells were diminished in the ORI-treated group. These anticancer effects of ORI maybe partly mediated by suppressing JAK2/STAT3 signaling pathway. These results highlight the potential effect of ORI on anti-angiogenesis and inducing vessel normalization roles of ORI, and probably provide optimum time point for the ORI therapy in conjunction with the chemoradiotherapy or immunotherapy.
肿瘤血管呈现出形态和功能异常。其成熟度和功能与结肠癌进展及治疗效果密切相关。目前仍缺乏直接证据证明冬凌草甲素（ORI）是否具有促进血管正常化的作用，而联合治疗可能会从中受益。我们建立了人结肠癌皮下异种移植模型。将动物分为对照组和ORI治疗组。采用免疫组织化学分析和TUNEL染色来评估增殖、凋亡和血管生成。运用蛋白质印迹分析来表征血管生成相关因子和JAK2/STAT3信号传导。然后，通过免疫荧光分析评估血管正常化和巨噬细胞重编程。结果显示，ORI明显抑制肿瘤生长，减少Ki67细胞数量和CD31微血管密度，同时增加TUNEL细胞数量。在ORI治疗组中，VEGF和bFGF蛋白的表达水平显著下调，而血管抑素和内皮抑素水平升高。此外，从第5天到第10天，ORI治疗显著促进了肿瘤血管的周细胞覆盖，在第7天周细胞覆盖率最高。在血管正常化的时间窗口内，ORI改善了肿瘤微环境的缺氧状态，下调了HIF-1a的表达。此外，在ORI治疗组中，CD206巨噬细胞减少。ORI的这些抗癌作用可能部分是通过抑制JAK2/STAT3信号通路介导的。这些结果突出了ORI在抗血管生成和诱导血管正常化方面的潜在作用，并可能为ORI与放化疗或免疫治疗联合应用提供最佳时间点。
