微小RNA-506对胰腺癌肿瘤相关巨噬细胞及肿瘤微环境的影响  

The effect of miR-506 on the tumor associated macrophages and tumor microenvironment of pancreatic ductal adenocarcinoma

作　　者：孙龙昊[1] 张杨[1] 陈岩[1] 梁晓宇[1] Sun Longhao;Zhang Yang;Chen Yan;Liang Xiaoyu(Department of General Surgery,Tianjin Medial University General Hospital,Tianjin 300052,China)

机构地区：[1]天津医科大学总医院普通外科,300052

出　　处：《中华肝胆外科杂志》2021年第12期928-931,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金(81702410);天津市自然科学基金(17JCQNJC11100);天津医科大学自然科学基金(2016KYZQ02)。

摘　　要：目的探索微小RNA-506(miR-506)对胰腺癌肿瘤相关巨噬细胞及肿瘤微环境的影响。方法使用Panc02细胞建立原位成瘤模型。将C57BL/6荷瘤小鼠分为miR-ctrl C57组和miR-506 C57组,每组15只,成瘤后分别通过腹腔注射二油酰磷脂酰胆碱(DOPC)脂质体包被的miR-ctrl(miR-ctrl/DOPC)或miR-506(miR-506/DOPC),测量肿瘤重量和体积,以流式细胞术检测肿瘤浸润性免疫细胞亚群变化,并监测小鼠生存期。将荷瘤裸鼠分为miR-ctrl nude组和miR-506 nude组,每组5只,成瘤后分别通过腹腔注射miR-ctrl/DOPC或miR-506/DOPC,用以排除免疫系统影响。将C57BL/6荷瘤小鼠分为清除对照组和巨噬细胞清除组,每组10只,成瘤后两组分别通过腹腔注射磷酸盐缓冲液(PBS)或氯膦酸二钠脂质体(Clo),用以排除巨噬细胞影响,2周后各组再分为miR-ctrl PBS组、miR-506 PBS组、miR-ctrl Clo组和miR-506 Clo组。结果miR-506 C57组小鼠的肿瘤体积(1.04±0.23)×103 mm 3和肿瘤重量(0.51±0.10)g均小于miR-ctrl C57组[(1.92±0.34)×103 mm 3和(0.99±0.19)g],M2/M1比例低于miR-ctrl C57组[(1.19±0.46)比(2.85±0.40)],差异具有统计学意义(P<0.01)。生存曲线显示,miR-506 C57组小鼠总生存期好于miR-ctrl C57组小鼠,差异具有统计学意义(P<0.01)。BALB/c裸鼠成瘤模型中miR-506 nude组小鼠的肿瘤体积和肿瘤重量与miR-ctrl nude组比较,差异无统计学意义(P>0.05)。C57BL/6荷瘤小鼠模型免疫微环境中miR-506 PBS组较miR-ctrl PBS组肿瘤调节性T细胞的比例更低[(4.70±1.86)%比(12.00±2.24)%],细胞毒性T细胞的比例更高[(10.54±1.89)%比(4.54±1.25)%],凋亡细胞毒性T细胞的比例更低[(14.00±4.00)%比(26.80±4.27)%],细胞毒性T细胞产生干扰素-γ的浓度更高[(89.60±14.69)比(28.00±13.69)ng/g],差异均具有统计学意义(P<0.05);但经过Clo清除巨噬细胞后,miR-506 Clo组和miR-ctrl Clo组上述指标差异均无统计学意义(P>0.05)。结论MiR-506通过巨噬细胞依赖的方式抑制胰腺癌的进展并ObjectiveTo explore the effect of miR-506 on the tumor associated macrophage(TAM)and tumor microenvironment(TME)of pancreatic ductal adenocarcinoma(PDAC).MethodsPanc02 cells were used to establish in situ tumorigenesis mouse model.Tumor bearing C57BL/6 mice were divided into miR-ctrl C57 group and miR-506 C57 group,with 15 mice in each group.miR-ctrl or miR-506 coated with DOPC liposomes(miR-ctrl/DOPC or miR-506/DOPC)were injected intraperitoneally.The tumor weight and volume were measured,the changes of tumor infiltrating immune cells were detected by flow cytometry and the survival time of mice was monitored.To eliminate the influence of immune system,tumor bearing nude mice were divided into miR-ctrl nude group and mir-506 nude group,with 5 mice in each group.miR-ctrl/DOPC or miR-506/DOPC were injected intraperitoneally.To eliminate the influence of macrophages,tumor bearing C57BL/6 mice were randomly divided into control group and macrophage clearance group and injected intraperitoneally with PBS or clodronate liposome(Clo),with 10 mice in each group.Then each group was randomly divided into two groups(miR-ctrl PBS group and miR-506 PBS group,miR-ctrl Clo group and miR-506 Clo group),miR-ctrl PBS group and miR-ctrl Clo group were injected with miR-ctrl/DOPC via intraperitoneal injection,miR-506 PBS group and miR-506 Clo group were injected with miR-506/DOPC.ResultsIn C57BL/6 mice model,the tumor volume(1.04±0.23)×103 mm 3 and tumor weight(0.51±0.10)g of miR-506 C57 group was lower than those in miR-ctrl C57 group(1.92±0.34)×103 mm 3 and(0.99±0.19)g,the overall survival of miR-506 C57 group was longer than that of miR-ctrl C57 group.The ratio of M2/M1 in miR-506 C57 group(1.19±0.46)was lower than that in miR-ctrl C57 group(2.85±0.40).In BALB/C nude mice model,the tumor volume and tumor weight showed no significant difference between miR-506 nude group and miR-ctrl nude group.Compared with the miR-ctrl PBS group,the proportion of regulatory T cells decreased[(4.70±1.86)%vs.(12.00±2.24)%],the proportio

关 键 词：微RNAS 胰腺肿瘤 巨噬细胞 肿瘤微环境 

分 类 号：R735.9[医药卫生—肿瘤]