文章摘要
黄 杰,胡 华,蒋 林.长链非编码RNA CBR3⁃AS1在骨肉瘤中的表达及意义[J].南京医科大学学报,2020,(1):56~61
长链非编码RNA CBR3⁃AS1在骨肉瘤中的表达及意义
Expression and significance of LncRNA CBR3⁃AS1 in osteosarcoma
投稿时间:2019-05-08  
DOI:10.7655/NYDXBNS20200111
中文关键词: 长链非编码RNA CBR3⁃AS1  表达  骨肉瘤  预后
英文关键词: LncRNA CBR3⁃AS1  expression  osteosarcoma  prognosis
基金项目:
作者单位
黄 杰 华中科技大学同济医学院附属武汉中心医院骨科湖北 武汉 430014 
胡 华 华中科技大学同济医学院附属武汉中心医院骨科湖北 武汉 430014 
蒋 林 华中科技大学同济医学院附属武汉中心医院骨科湖北 武汉 430014 
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中文摘要:
      目的:探讨长链非编码RNA CBR3?AS1(LncRNA CBR3?AS1)在骨肉瘤患者中的表达及意义。方法:回顾性收集2012年1月—2014年1月本院收治的66例骨肉瘤患者组织标本和临床资料,qRT?PCR法检测组织中LncRNA CBR3?AS1表达情况,并将患者分为高表达组和低表达组,卡方检验分析LncRNA CBR3?AS1表达与临床参数的关系,Kaplan?Meier 法比较LncRNA CBR3?AS1的表达与总生存率的关系。将U?2 OS细胞系分成LncRNA CBR3?AS1沉默表达组(si?CBR3?AS1组)和阴性对照组(si?NC组),经LipofectamineTM 3000分别转染LncRNA CBR3?AS1 siRNA和si?NC序列,MTT和集落形成试验测定细胞增殖,细胞迁移和侵袭试验测定细胞迁移和侵袭能力,流式细胞术测定细胞凋亡。qRT?PCR和Western blot法检测羧基还原酶3(CBR3)的表达。结果:LncRNA CBR3?AS1在骨肉瘤组织和细胞系中高表达(P < 0.001)。LncRNA CBR3?AS1高表达与Enneking分期(P < 0.001)、远处转移(P=0.004)和组织学分级(P=0.036)显著相关;LncRNA CBR3?AS1高表达与骨肉瘤患者总生存率呈负相关(P < 0.01),是骨肉瘤患者的不良预后因素(HR=1.558,95%CI:1.041~2.641,P=0.013)。si?CBR3?AS1组细胞增殖﹑集落形成能力、细胞迁移和侵袭能力显著低于si?NC组,si?CBR3?AS1组细胞凋亡率高于si?NC组。LncRNA CBR3?AS1和CBR3 mRNA表达呈正相关(r=0.44,P=0.036),si?CBR3?AS1组CBR3 mRNA和蛋白表达量显著低于si?NC组(P < 0.001)。结论:LncRNA CBR3?AS1在骨肉瘤中高表达,LncRNA CBR3?AS1下调表达抑制细胞增殖、迁移及侵袭,促进凋亡,是影响骨肉瘤预后的独立危险因素。
英文摘要:
      Objective:To investigate the expression and significance of long non?coding RNA CBR3?AS1(LncRNA CBR3?AS1)in osteosarcoma tissue and cell lines. Methods:The pathological tissue samples and clinical data of 66 patients with osteosarcoma diagnosed and treated in our hospital from January 2012 to January 2014 were retrospectively collected. The expression of LncRNA CBR3?AS1 in pathological tissues was detected by qRT?PCR. The patients were divided into the high expression group and the low expression group. The correlation of the expression of LncRNA CBR3?AS1 and clinical parameters was analyzed by Chi?square test. Kaplan?Meier survival analysis was used to compare the survival rate of osteosarcoma patients with high or low expression of LncRNA CBR3?AS1. U?2 OS cell lines were divided into the LncRNA CBR3?AS1 silent expression group(si?CBR3?AS1 group)and the negative control group(si?NC group). LncRNA CBR3?AS1 siRNA and si?NC sequences were transfected by LipofectamineTM 3000,respectively. Cell proliferation,cell migration and invasion were measured by MTT and colony formation assay. Apoptotic capacity was measured by cytometry. Expression of carboxyl reductase 3(CBR3)were detected by qRT?PCR and Western blot. Results:LncRNA CBR3?AS1 were highly expressed in osteosarcoma tissues and cell lines(P < 0.001). LncRNA CBR3?AS1 high expression were significantly correlated with Enneking staging(P < 0.001),distant metastasis(P=0.004)and histological grade(P=0.036). LncRNA CBR3?AS1 high expression was negatively correlated with overall survival rate in osteosarcoma patients(P < 0.01), it was an independent prognostic factor for osteosarcoma patients(HR=1.558,95%CI:1.041?2.641,P=0.013). Cell proliferation and colony formation ability,as well as cell migration and invasion ability in the si?CBR3?AS1 group were significantly lower than those in the si?NC group,and cell apoptosis rate in the si?CBR3?AS1 group was higher than that in the si?NC group. The expression of LncRNA CBR3?AS1 and CBR3 was positively correlated(r=0.44,P=0.036). The expression of CBR3 protein in the si?CBR3?AS1 group was significantly lower than that in the si?NC group(P < 0.001). Conclusion:Downregulated expression of LncRNA CBR3?AS1 inhibits cell proliferation,migration and invasion,and promotes apoptosis,which is an independent risk factor affecting the prognosis of osteosarcoma.
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