摘要
目的:探讨IL-37对肾细胞癌细胞生物学行为的影响和潜在机制。方法:RT-qPCR检测肾细胞癌组织中IL-37mRNA和miR-106b-5p表达。Western blot检测肾细胞癌组织中10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)蛋白表达。将肾细胞癌细胞786-0分为对照组、IL-37(10、50、100 ng/ml)组、anti-miR-NC组、anti-miR-106b-5p组、IL-37+miR-NC组、IL-37+miR-106b-5p组。采用MTT法、平板克隆实验、划痕愈合实验、流式细胞术检测786-0细胞增殖、迁移和凋亡能力。荧光素酶实验检测miR-106b-5p与PTEN的靶向关系。结果:肾细胞癌组织中miR-106b-5p表达量显著升高(P<0.05),IL-37 mRNA和PTEN蛋白表达量显著降低(P<0.05)。与对照组比较,IL-37(10、50、100 ng/ml)组细胞活力、集落形成数、迁移距离、miR-106b-5p表达显著降低(P<0.05),凋亡率、PTEN蛋白表达显著升高(P<0.05)。与anti-miR-NC组比较,anti-miR-106b-5p组细胞活力、集落形成数、迁移距离显著降低(P<0.05),凋亡率、PTEN蛋白表达显著升高(P<0.05)。与IL-37+miR-NC组比较,IL-37+miR-106b-5p组细胞活力、集落形成数、迁移距离显著升高(P<0.05),凋亡率、PTEN蛋白表达显著降低(P<0.05)。PTEN是miR-106b-5p的靶基因。结论:外源性IL-37可抑制肾细胞癌细胞的增殖和迁移,诱导细胞凋亡,其抗肿瘤机制可能是通过抑制miR-106b-5p/PTEN途径发挥作用。
Objective:To investigate the effect of IL-37 on the biological behavior of renal cell carcinoma cells and further to explore its potential mechanism.Methods:The expressions of IL-37 mRNA and miR-106b-5p in renal cell carcinoma tissues were measured using RT-qPCR.The expression of phosphatase and tensin homolog deleted on chromosome 10(PTEN)protein in renal cell carcinoma tissues were detected by Western blot.Renal cell carcinoma 786-0 cells were divided into control group,IL-37(10,50,100 ng/ml)groups,anti-miR-NC group,anti-miR-106b-5p group,IL-37+miR-NC group and IL-37+miR-106b-5p group.MTT method,plate cloning assay,scratch healing assay and flow cytometry were applied to detect the viability,colony formation migration and apoptosis of 786-0 cell respectively.Luciferase experiment was performed to confirm the targeting relationship between miR-106b-5p and PTEN.Results:The expression of miR-106b-5p in renal cell carcinoma tissue was significantly increased(P<0.05),while the expressions of IL-37 mRNA and PTEN protein were significantly decreased(P<0.05).Compared with the control group,the cell viability,colony formation number,migration distance,miR-106b-5p expression in IL-37(10,50,100 ng/ml)groups were significantly decreased(P<0.05),apoptosis rate and PTEN protein expression were significantly increased(P<0.05).Compared with the anti-miRNC group,the cell viability,colony formation number and migration distance in anti-miR-106b-5p group were significantly reduced(P<0.05),and apoptosis rate and PTEN protein expression were significantly increased(P<0.05).Compared with IL-37+miR-NC group,the cell viability,colony formation number,and migration distance in IL-37+miR-106b-5p group were significantly increased(P<0.05),and apoptosis rate and PTEN protein expression were significantly decreased(P<0.05).PTEN is the target gene of miR-106b-5p.Conclusion:Exogenous IL-37 can inhibit cell proliferation and migration,and induce cell apoptosis in renal cell carcinoma,whose anti-tumor mechanism may be achieved by inhibiting mi
作者
顾鹏
陶维雄
彭伟
魏世平
GU Peng;TAO Weixiong;PENG Wei;WEI Shiping(Department of Urology,Changjiang Shipping General Hospital,Wuhan 430000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2023年第8期1694-1699,共6页
Chinese Journal of Immunology
