四连接素通过腺苷酸活化蛋白激酶/血管内皮生长因子轴抑制前列腺癌的增殖、迁移  

C-type lectin domain family 3 member B inhibits prostate cancer proliferation and migration through the adenosine monophosphate-actived protein kinase/vascular endothelial growth factor axis

作　　者：李冠儒[1] 张二伟[1] 袁博[1] 王启[1] Li Guanru;Zhang Erwei;Yuan Bo;Wang Qi(Department of Urology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第一附属医院泌尿外科,郑州450000

出　　处：《中华实验外科杂志》2025年第1期79-82,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探索四连接素(CLEC3B)对前列腺癌增殖、迁移的影响及其作用机制。方法分析肿瘤基因组图谱(TCGA)数据库中497例前列腺癌和52例正常前列腺组织中基因CLEC3B的表达差异。将人源的前列腺癌细胞系22RV1和DU145分为对照组和实验组,分别转染空质粒和CLEC3B质粒,转染2 d后,采用平板克隆形成实验、细胞计数试剂盒(CCK-8)实验和5-乙炔基-2’脱氧尿嘧啶核苷(EdU)染色实验验证细胞的增殖能力;采用Transwell实验和划痕愈合实验验证细胞的迁移能力,采用蛋白质印迹法探索CLEC3B与AMP依赖的蛋白激酶(AMPK)/血管内皮生长因子(VEGF)轴的靶向关系。两实验组间比较采用独立样本t检验,多实验组间比较采用方差分析。结果实验组细胞EdU着色细胞比例低于对照组[22RV1:(50.333±4.726)%比(80.333±2.082)%,t=10.062,P<0.05;DU145:(30.333±3.512)%比(64.000±5.000)%,t=9.544,P<0.05]、培养96 h后吸光度值低于对照组(22RV1:1.144±0.183比1.724±0.113,F=46.827,P<0.05;DU145:1.473±0.140比2.170±0.179,F=61.480,P<0.05)、克隆形成数低于对照组(22RV1:114.333±17.898比279.000±29.816,t=8.202,P<0.05;DU145:85.333±12.097比168.333±11.015,t=8.787,P<0.05)、划痕愈合率低于对照组(22RV1:26.333±5.033比84.333±9.866,t=9.070,P<0.05;DU145:51.000±6.000比105.333±12.897,t=6.616,P<0.05)、迁移细胞数低于对照组[22RV1:(11.000±6.000)%比(26.667±4.509)%,t=3.615,P<0.05;DU145:(26.667±5.132)%比(70.667±3.512)%,t=12.256,P<0.05]。同时,CLEC3B能显著增加p-AMPK蛋白的表达(22RV1:0.515±0.076比0.168±0.069,t=5.922,P<0.05;DU145:0.376±0.081比0.080±0.018,t=6.156,P<0.05)、降低VEGF蛋白的表达(22RV1:0.135±0.029比0.524±0.058,t=10.353,P<0.05;DU145:0.103±0.008比0.434±0.034,t=16.283,P<0.05)。结论CLEC3B能够通过AMPK/VEGF轴抑制前列腺癌细胞的增殖和迁移。Objective To investigate the impact of C-type lectin domain family 3 member B(CLEC3B)on the proliferation and migration of prostate cancer cells and its underlying mechanisms.Methods Differential expression of the CLEC3B gene was analyzed in 497 prostate cancer samples and 52 normal prostate tissues from the the cancer genome atlas(TCGA)database.Human prostate cancer cell lines 22RV1 and DU145 were divided into control and experimental groups,transfected with empty plasmids and CLEC3B plasmids,respectively.After 2 days of transfection,the proliferative capacity of the cells was assessed using colony formation,cell counting kit-8(CCK-8),and 5-Ethynyl-2′-deoxyuridine(EdU)staining assays.Transwell and scratch wound healing assays were used to evaluate cell migration capacity.Western blotting was employed to explore the relationship between CLEC3B and the adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)/vascular endothelial growth factor(VEGF)axis.Statistical analysis was conducted using independent sample t-tests for comparisons between two groups and ANOVA for multiple group comparisons.Results The proportion of EdU-positive cells in the experimental group was significantly lower than that in the control group[22RV1:(50.333±4.726)%vs.(80.333±2.082)%,t=10.062,P<0.05;DU145:(30.333±3.512)%vs.(64.000±5.000)%,t=9.544,P<0.05].After 96 h of culture,the absorbance(A)density in the experimental group was lower than in the control group(22RV1:1.144±0.183 vs.1.724±0.113,F=46.827,P<0.05;DU145:1.473±0.140 vs.2.170±0.179,F=61.480,P<0.05).Colony formation number was also reduced(22RV1:114.333±17.898 vs.279.000±29.816,t=8.202,P<0.05;DU145:85.333±12.097 vs.168.333±11.015,t=8.787,P<0.05).Scratch wound healing rates were significantly lower in the experimental group(22RV1:26.333±5.033 vs.84.333±9.866,t=9.070,P<0.05;DU145:51.000±6.000 vs.105.333±12.897,t=6.616,P<0.05),and the number of migrating cells was also significantly lower[22RV1:(11.000±6.000)%vs.(26.667±4.509)%,t=3.615,P<0.05;DU145:(26.667±

关 键 词：前列腺癌 四连接素 腺苷酸活化蛋白激酶 血管内皮生长因子 

分 类 号：R737.25[医药卫生—肿瘤]