约瑟芬结构域含蛋白2对肾透明细胞癌的增殖和迁移能力的影响
Impact of Josephin domain-containing protein 2 on proliferation and migration of clear cell renal cell carcinoma
王豪 1朱卫 2谢冲 2付琪奇 1万梓宇 1余力 3李艳松 3胡祎舜 3刘敏豪 3汤晨昊 3彭袈豪 3廖义翔 3彭建平1
作者信息
- 1. 武汉大学中南医院泌尿外科,武汉 430071
- 2. 江陵县人民医院泌尿外科,江陵 434100
- 3. 荆州市中心医院长江大学附属荆州医院泌尿外科,荆州 434020
- 折叠
摘要
目的 探讨约瑟芬结构域含蛋白2(JOSD2)的表达对肾透明细胞癌增殖与迁移的影响.方法 在Timer、UALCAN,数据库中分析JOSD2在肾癌中的差异表达和预后(P＜0.05).在Caki-1细胞系中使用成簇的规律间隔短回文重复序列(CRISPR)/Cas9技术导入shRNA敲低JOSD2.设置对照组和稳定JOSD2敲低组(sh1、sh2),通过细胞计数试剂盒(CCK-8)增殖实验、平板克隆实验、Transwell实验分别观察对细胞增殖、迁移能力的影响.采用独立样本t检验进行组间比较.结果 生信数据库发现JOSD2在多种肿瘤中异常高表达,其中包括肾透明细胞癌[28.707(22.825～36.424)比 20.433(17.507～23.548),P＜0.05].在敲低 JOSD2 后,CCK-8 结果显示,JOSD2敲低后细胞增殖能力明显低于对照组(3.11±0.08比2.12±0.13,t=14.64,P＜0.05;3.10±0.08比2.18±0.20,t=10.26,P＜0.05).平板克隆形成实验结果显示,JOSD2敲低后细胞集落形成能力显著低于对照组[(365.0±34.6)个比(217.0±22.0)个,i=6.948 P＜0.05;(365.0± 34.6)个 比(186.0±38.2)个,t=7.216,P＜0.05].Transwell实验显示,JOSD2敲低后细胞迁移能力显著低于对照组[(1 234.5±85.1)个比(689.8±58.7)个,t=10.540,P＜0.05;(1 234.5± 85.1)个 比(635.0±112.3)个,t=6.967,P＜0.05].结论 JOSD2 在肾癌中表达上调,敲低 JOSD2能抑制肾癌细胞增殖和迁移.
Abstract
Objective To investigate the effect of Josephin domain-containing protein 2(JOSD2)expression on the proliferation and migration of renal clear cell carcinoma(RCCC).Methods Differential expression of JOSD2 in RCCC,RCCC stage and prognosis were analyzed in Timer,UALCAN databases(P＜0.005).The shRNA knockdown of JOSD2 was introduced in Caki-1 cell lines using regular interval short palindromic repeat clusters(CRISPR)/Cas9 technology.Groups were set up as Caki-1 control,and Caki-1 JOSD2 knockdown experimental groups(SHI,SH2).The effects of JOSD2 on cell proliferation and migration were assessed using the cell counting kit-8(CCK-8)assay,plate colony formation assay,and Transwell assay,respectively.A comparison between groups was conducted using an independent sample t-test.Results The bioinformatics database revealed a significant upregulation of JOSD2 in various canc-ers,including RCCC[28.707(22.825-36.424)vs.20.433(17.507-23.548),P＜0.05].After knock-down of JOSD2,the CCK-8 results demonstrated a significant reduction in cell proliferation ability(3.11± 0.08 vs.2.12±0.13,t=14.64,P＜0.05;3.1±0.08 vs.2.18±0.20,t=10.26,P＜0.05).Plate colony formation assay showed that the colony formation ability of JOSD2 knockdown cells was significantly decreased(365.0±34.6 vs.217.0±22.0,t=6.948 P＜0.05;365.0±34.6 vs.186.0±38.2,t=7.216,P＜0.05).Transwell assay showed that the cell migration ability was significantly decreased after JOSD2 knockdown[(1 234.5±85.1)vs.(689.8±58.7),t=10.540,P＜0.05;(1 234.5±85.1)vs.(635.0±112.3),t=6.967,P＜0.05].Conclusion The expression of JOSD2 is upregulated in RCCCs,and knockdown of JOSD2 can inhibit the proliferation and migration of RCCCs.
关键词
约瑟芬结构域含蛋白2/肾透明细胞癌Key words
Josephin domain-containing protein 2/Clear cell carcinoma of kidney引用本文复制引用
出版年
2024
NETL
NSTL
万方数据