Zhang Hongbo, Yan Mengyu, Zhang Jiandong, Sun Peiwang, Wang Rui, Guo Yuanyuan
Department of Urology, First Affiliated Hospital of Bengbu Medical University, Bengbu 233040, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 Jul 20;45(7):1513-1518. doi: 10.12122/j.issn.1673-4254.2025.07.18.
To investigate the inhibitory effect of pirfenidone (PFD) on growth of bladder cancer xenograft and its regulatory effect on Treg cells in tumor-bearing mice.
Thirty-two C57BL/6 mice bearing ectopic bladder tumors were randomized into control and PFD groups (=16). In PFD group, PFD was administered orally at the daily dose of 500 mg/kg, and tumor growth and survival of the mice were monitored. After treatment for 21 days, the tumors and vital organs were harvested for analysis. Immunohistochemistry was used to assess CD3, CD4, CD8, and FOXP3 expressions in the tumors. Flow cytometry and RT-qPCR were used to analyze the percentage of CD4⁺CD25⁺FOXP3⁺ Treg cells and IL-2, IL-10, and IL-35 expressions in the tumors and spleens; organ damage of the mice was examined with HE staining.
Compared with the control group, the PFD-treated mice exhibited significantly lower tumor growth rate with smaller tumor volumes at day 21, along with improved survival at day 28. Immunohistochemistry revealed no significant differences in the infiltration of CD3⁺ and CD8⁺ cells between the two groups, but the percentages of CD4⁺ and FOXP3⁺ cells were significantly lower in the tumors of PFD-treated mice. Flow cytometric analysis confirmed a decrease in CD4⁺CD25⁺FOXP3⁺ Treg cells in the tumors from PFD-treated mice, which also had reduced expression levels of IL-2, IL-10 and IL-35 mRNAs in the tumors. No significant differences were found in Treg cell populations or cytokine expressions in the spleen tissues between the two groups. HE staining showed obvious organ damage in neither of the groups.
PFD inhibits bladder cancer growth and enhances survival of tumor-bearing mice possibly by suppressing Treg cells in the tumor microenvironment.
探讨吡非尼酮（PFD）对膀胱癌异种移植物生长的抑制作用及其对荷瘤小鼠体内调节性T细胞（Treg细胞）的调控作用。
将32只异位膀胱肿瘤荷瘤C57BL/6小鼠随机分为对照组和PFD组（每组16只）。PFD组小鼠按500 mg/kg的日剂量口服PFD，监测小鼠肿瘤生长及生存情况。治疗21天后，采集肿瘤及重要脏器进行分析。采用免疫组织化学法评估肿瘤组织中CD3、CD4、CD8及叉头框蛋白3（FOXP3）的表达。采用流式细胞术和逆转录-定量聚合酶链反应（RT-qPCR）分析肿瘤组织和脾脏中CD4⁺CD25⁺FOXP3⁺ Treg细胞百分比以及白细胞介素-2（IL-2）、白细胞介素-10（IL-10）和白细胞介素-35（IL-35）的表达；通过苏木精-伊红（HE）染色检查小鼠的器官损伤情况。
与对照组相比，PFD治疗组小鼠在第21天时肿瘤生长速率显著降低，肿瘤体积更小，且在第28天时生存率提高。免疫组织化学显示，两组间CD3⁺和CD8⁺细胞浸润无显著差异，但PFD治疗组小鼠肿瘤组织中CD4⁺和FOXP3⁺细胞百分比显著降低。流式细胞术分析证实，PFD治疗组小鼠肿瘤组织中CD4⁺CD25⁺FOXP3⁺ Treg细胞减少，肿瘤组织中IL-2、IL-10和IL-35 mRNA表达水平也降低。两组脾脏组织中Treg细胞群体或细胞因子表达无显著差异。HE染色显示两组均无明显器官损伤。
PFD可能通过抑制肿瘤微环境中的Treg细胞来抑制膀胱癌生长并提高荷瘤小鼠的生存率。
