南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (2): 210-216.doi: 10.12122/j.issn.1673-4254.2024.02.02

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吡非尼酮联合PD-L1抑制剂抑制小鼠异位膀胱肿瘤的生长

陈守峰，张舒超，樊伟林，孙 巍，刘贝贝，刘建民，郭园园   

  1. 蚌埠医学院第一附属医院泌尿外科，安徽 蚌埠 233040
  • 发布日期:2024-03-13

Efficacy of combined treatment with pirfenidone and PD-L1 inhibitor in mice bearing ectopic bladder cancer xenograft

CHEN Shoufeng, ZHANG Shuchao, FAN Weilin, SUN Wei, LIU Beibei, LIU Jianmin, GUO Yuanyuan   

  1. Department of Urology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233040, China
  • Published:2024-03-13

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摘要： 目的 通过小鼠肿瘤模型观察吡非尼酮（PFD）联合程序性死亡受体-配体1（PD-L1）抑制剂对膀胱癌的疗效及对免疫微环境的调控作用。方法 构建C57BL/6小鼠异位膀胱肿瘤模型共40只，根据不同处理随机分为4组（10只/组）：对照组、PD-L1抑制剂组、PFD组、联合治疗组。对照组：建立肿瘤模型正常饮食；PD-L1抑制剂组：建模后腹腔每3 d按12.5 mg/kg注射PD-L1抑制剂；PFD组：建模后每天按500 mg/kg口服PFD；联合治疗组：建模后PFD及PD-L1抑制剂按上述剂量联合应用。对比各组小鼠生存率和肿瘤生长速度，药物干预21 d后留取肿瘤组织及小鼠血清，免疫组化检测肿瘤组织中CD3、CD8、CD45、E-cadherin及 N-cadherin的表达；免疫荧光观察骨髓来源抑制细胞（MDSCs）表达；生化分析小鼠血液中谷丙转氨酶（ALT）、谷草转氨酶（AST）、血尿素氮（BUN）、肌酐（CRE）及乳酸脱氢酶（LDH-L）的水平。结果 与对照组相比，PD-L1抑制剂组和PFD组小鼠肿瘤相对生长速率及21 d肿瘤体积均减小（P<0.05），联合治疗组小鼠更加显著（P<0.05）。免疫组化及免疫荧光结果显示，PD-L1抑制剂组与PFD组小鼠肿瘤组织E-cadherin表达增加，N-cadherin表达降低（P<0.05）。CD3+T细胞、CD8+T细胞、CD45+T细胞数量较对照组增加，而Ly-6G+CD11b+MDSCs细胞减少，联合组变化更加明显（P<0.05）。生化分析结果显示，各组小鼠血清ALT、AST、BUN、CRE及LDH-L水平无明显差异（P>0.05）。 结论 吡非尼酮联合PD-L1抑制剂可显著抑制膀胱癌的进展，其效应可能是通过调节肿瘤微环境，抑制肿瘤细胞的上皮间质转化来实现的。

关键词: 吡非尼酮；PD-L1；膀胱癌；免疫微环境；髓系抑制细胞

Abstract: Objective To assess the efficacy of pirfenidone combined with PD-L1 inhibitor for treatment of bladder cancer in a mouse model and its effect on tumor immune microenvironment modulation. Methods Forty C57BL/6 mouse models bearing ectopic human bladder cancer xenografts were randomized into control group, PD-L1 inhibitor group, pirfenidone group and combined treatment group (n=10). After successful modeling, PD-L1 inhibitor treatment was administered via intraperitoneal injection at 12.5 mg/kg every 3 days, and oral pirfenidone (500 mg/kg) was given on a daily basis. The survival rate of the mice and tumor growth rate were compared among the 4 groups. The expressions of CD3, CD8, CD45, E-cadherin and N-cadherin in the tumor tissues were detected with immunohistochemistry after the 21-day treatment, and bone marrow-derived suppressor cells (MDSCs) were observed with immunofluorescence staining; serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN), creatinine (CRE) and lactate dehydrogenase (LDH-L) were analyzed using an automated biochemical analyzer. Results Treatment with PD- L1 inhibitor and pirfenidone alone both significantly decreased tumor growth rate and tumor volume at 21 days (P<0.05), but the combined treatment produced an obviously stronger inhibitory effect (P<0.05). PD- L1 inhibitor and pirfenidone alone significantly increased E- cadherin expression and decreased N-cadherin expression in the tumor tissue (P<0.05). The two treatments both significantly increased the percentage of CD3+, CD8 and CD45+ T cells and decreased the percentage of Ly-6G+CD11b+MDSCs in the tumor tissue, and these changes were more obvious in the combined treatment group (P<0.05). No significant differences were found in serum ALT, AST, BUN, CRE or LDH- L levels among the 4 groups (P>0.05). Conclusion Combined treatment with pirfenidone and PD-L1 inhibitor significantly inhibits the progression of bladder cancer in mice possibly by regulating tumor immune microenvironment and inhibiting epithelial-mesenchymal transition of the tumor cells.

Key words: pirfenidone; PD-L1; bladder cancer; immune microenvironment; bone marrow-derived suppressor cells