外泌体源性长链非编码RNA同源框基因A-反义转录本3通过吸附微小RNA-29b-3p调节前列腺癌去势抵抗的作用机制
The mechanism of cancer derived exosomal homeobox gene a-antisense transcript 3 regulating castration resistance of prostate cancer by sponging microRNA-29b-3p
蔡启亮 1杨先瑞 1姚智力 1侯泽楷 1王晨宇 1崔荣昊 1杨思维 1王准 1李刚 1权昌益 1牛远杰1
作者信息
- 1. 天津医科大学第二医院泌尿外科,天津 300211
- 折叠
摘要
目的 探讨外泌体来源长链非编码RNA(lncRNA)同源框基因A-反义转录本3(HOXA-AS3)对前列腺癌(PCa)去势抵抗的作用机制.方法 建立PC-3与LNCaP共培养体系,用双荧光素酶报告基因检测、荧光定量聚合酶链反应(PCR)和挽救实验等验证HOXA-AS3参与PCa去势抵抗和进展的机制.采用动物实验和人PCa组织切片,探究HOXA-AS3在由雄激素依赖性向非依赖性的转化过程中的作用.采用两样本t检验和单因素方差分析(One-Way ANOVA)进行统计分析.结果 ASO-NC转染组LNCaP细胞侵袭数量显著低于ASO-miR-29b-3p转染组[(116.00±5.10)个比(146.33±4.78)个,t=6.135,P＜0.05],NC 转染 PC-3 组显著高于 miR-29b-3p mimic 转染组[(394.33±11.95)个比(111.67±7.41)个,t=28.425,P＜0.05].LNCaP-HOXA-AS3-exo 中HOXA-AS3 显著高于 LNCaP-exo(1.46±0.08 比 1.01±0.02,t=7.634,P＜0.05),而 PC-3-HOXA-AS3-ASO-exo 中 HOXA-AS3 显著低于 PC-3-exo(0.53±0.06 比 1.00±0.09,t=5.943,P＜0.05).转染miR-29b-3p-mimic组的野生型HOXA-AS3、野生型STAT3-3和野生型Mcl-1组的细胞荧光素酶活性低于对照组(1.00±0.04 比 0.51±0.02、1.00±0.02 比 0.56±0.04、1.00±0.02 比 0.57±0.02,t=18.058、15.857、26.081,P＜0.01).同时转染 pcDNA3.1-STAT3 和 Mcl-1-promotor 载体组免疫荧光强度高于对照组(1.72±0.05 比 1.00±0.03,t=18.717,P＜0.05),并且 cytochrome c 和 Caspase-9 的表达低于对照组(0.46±0.01 比 0.81±0.01、0.28±0.01 比 0.59±0.01,t=92.492、97.212,P＜0.01).结论 HOXA-AS3 通过吸附 miR-29b-3p 调节 STAT3/Mcl-1/cytochrome c/Caspase-9 轴,调节PCa的去势抵抗.
Abstract
Objective To investigate the role of exosomal long no-coding RNA(lncRNA)ho-meobox gene a-antisense transcript 3(HOXA-AS3)in castration resistance and progression of prostate cancer(PCa).Methods Construct a co-culture system between PC-3 and LNCaP.The dual luciferase re-porter gene detection,PCR(polymerase chain reaction)and Rescue were conducted to verify the role of ex-osomes in castration resistance and progression of PCa.Animal experiments and human prostate cancer tis-sue sections were conducted to investigate the role of HOXA-AS3 in the transition from androgen dependent prostate cancer(ADPC)to castration-resistant prostate cancer(CRPC).Two sample t-test or One-Way ANOVA was used for statistically analysis.Results The number of invasive cells in LNCaP containing ASO-NC group was significantly less than that in LNCaP containing miR-29b-3p-ASO group(116.00±5.10 vs.146.33±4.78,t=6.135,P＜0.05).The number of invasive cells in PC-3 containing mimic-NC group was significantly higher than that in PC-3 containing miR-29b-3p group(394.33±11.95 vs.111.67±7.41,t=28.425,P＜0.05).The content of HOXA-AS3 in LNCaP-HOXA-AS3-exo group is significantly higher than that in LNCaP-exo group(1.46±0.08 vs.1.01±0.02,t=7.634,P＜0.05).The content of HOXA-AS3 in PC-3-HOXA-AS3-ASO-exo group is significantly higher than that in PC-3-exo group(0.53±0.06 vs.1.00±0.09,t=5.943,P＜0.05).The overexpressed miR-29b-3p significantly reduced the luciferase intensity of HOXA-AS3-3'UTR wide type,STAT3-3'UTR wide type and Mcl-1-3'UTR wide type(1.00±0.04 vs.0.51±0.02,1.00±0.02 vs.0.56±0.04,1.00±0.02 vs.0.57±0.02,t=18.058,15.857,26.081,P＜0.01).The relative luciferase intensity of the group containing with pcDNA3.1-STAT3 and Mcl-1-promotor was significantly higher than that of the controlled group(1.72±0.05 vs.1.00±0.03,t=18.717,P＜0.05),and the expression of cytochrome c and caspase-9 was significantly less than that of the controlled group(0.81±0.01 vs.0.46±0.01,0.59±0.01 vs.0.28±0.01,t=92.492,97.212,P＜0.01).Conclusion HOXA-AS3 contributes to the castration resistance of PCa via sponging miR-29b-3p and regulating STAT3/Mcl-1/cytochrome c/caspase-9 axis.
关键词
前列腺癌/外泌体/反义转录本/长链非编码RNAKey words
Prostate cancer/Exosome/Antisense Transcript/Long non-coding RNA引用本文复制引用
基金项目
天津市杰出青年科学基金(23JCJQJC00080)
天津市泌尿外科研究所人才资助计划(MYSRC202305)
天津市卫生健康高层次人才培养工程(TJSQNYXXR-D2-158)
天津市教委科研计划项目(2022ZD068)
天津市科技计划项目(21JCYBJC01470)
天津市科技计划项目(21JCQNJC01790)
天津市卫生健康科技项目(TJWJ202021QN033)
出版年
2024
NETL
NSTL
万方数据