敲减USP39基因抑制人膀胱癌细胞增殖、促进细胞凋亡  

Knockdown of USP39 gene inhibits the proliferation of human bladder cancer cells and promotes cell apoptosis

作　　者：李晶[1] 屈卫星[1] 程永毅[1] LI Jing;QU Weixing;CHENG Yongyi(Department of Urology,Shaanxi Provincial People's Hospital,Shaanxi Xi'an 710068,China)

机构地区：[1]陕西省人民医院泌尿外科,陕西西安710068

出　　处：《现代肿瘤医学》2022年第5期791-796,共6页Journal of Modern Oncology

摘　　要：目的:探讨慢病毒介导的RNA干扰敲减泛素特异性肽酶39(ubiquitin specific peptidase 39,USP39)对人膀胱癌细胞生长和细胞凋亡的影响,为膀胱癌的治疗提供新思路。方法:RT-qPCR确定5637和T24细胞系中USP39 mRNA的表达;设计合成针对USP39的特异性shRNA,利用脂质体转染膀胱癌细胞后,RT-qPCR和Western blotting检测shRNA抑制效率;通过Celigo细胞计数测定、MTT法和集落形成能力检测细胞的增殖能力和集落形成能力;流式细胞术检测细胞凋亡;Western blotting检测凋亡标记物C-PARP1、p53、p-p53和C-Caspase-3的表达。结果:通过RT-qPCR和Western blotting分析确定,shRNA敲减后细胞中USP39的mRNA和蛋白质表达水平显著降低。通过MTT分析、集落形成分析和流式细胞术分析证实,USP39的敲减显著降低了膀胱癌细胞的增殖和集落形成能力,并诱导了细胞凋亡。此外,Western blotting分析表明,USP39基因敲减可能通过上调膀胱癌5637细胞中p53、p-p53、C-PARP1和C-Caspase-3的表达诱导细胞凋亡。结论:敲减USP39可通过激活Caspase级联反应和上调p53的表达来抑制膀胱癌细胞的增殖。这些数据可为开发USP39作为抗膀胱癌的潜在分子靶标提供实验依据。USP39可能是针对膀胱癌靶向治疗的新型生物标志物,需要进一步研究。Objective:To investigate the effect of lentivirus-mediated RNA interference knockdown ubiquitin specific peptidase 39(USP39)on the growth and apoptosis of human bladder cancer cells,and to provide new ideas for bladder cancer treatment.Methods:RT-qPCR determined the expression of USP39 mRNA in 5637 and T24 cell lines.Specific shRNA for USP39 was designed and synthesized.Bladder cancer cells were transfected with liposomes.RT-qPCR and Western blotting was used to detect the shRNA inhibition efficiency.Celigo cell counting assay,MTT method and colony-forming ability were used to detect the proliferation and colony-forming ability of cells.Flow cytometry was used to detect apoptosis.Western blotting was used to detect the expression of apoptosis markers C-PARP1,p53,p-p53 and C-Caspase-3.Results:It was determined by RT-qPCR and Western blotting that the expression levels of USP39 mRNA and protein in USP39 knockdown cells were significantly reduced.Through MTT analysis,colony formation analysis and flow cytometry analysis,it was confirmed that the knockdown of USP39 significantly reduced the proliferation and colony formation ability of bladder cancer cells,and induced apoptosis.In addition,Western blotting analysis showed that USP39 gene knockdown may induce apoptosis by up-regulating the expression of p53,p-p53,C-PARP1 and C-Caspase-3 in bladder cancer 5637 cells.Conclusion:Knockdown of USP39 can inhibit the proliferation of bladder cancer cells by activating the Caspase cascade and up-regulating the expression of p53.These data can provide experimental basis for the development of USP39 as a potential molecular target against bladder cancer.USP39 may be a new biomarker for targeted therapy of bladder cancer,and further research is needed.

关 键 词：泛素特异性肽酶39 膀胱癌 增殖 凋亡 RNA干扰 

分 类 号：R737.14[医药卫生—肿瘤]