长链非编码RNA B3GALT5-AS1抑制非小细胞肺癌A549细胞增殖、侵袭及体内致瘤性  

Long Non-coding RNA B3GALT5-AS1 Inhibits Proliferation,Invasion and Tumorigenicity of Non-small Cell Lung Cancer A549 Cells

作　　者：彭强 赵慧 姚秋英 PENG Qiang;ZHAO Hui;YAO Qiuying(Department of Pathology,Yiyang Central Hospital,Yiyang,Hunan,413099,China;Department of Oncology,Yiyang Central Hospital,Yiyang,Hunan,413099,China)

机构地区：[1]益阳市中心医院病理科,湖南省益阳市413099 [2]益阳市中心医院肿瘤科,湖南省益阳市413099

出　　处：《医学分子生物学杂志》2024年第5期425-431,共7页Journal of Medical Molecular Biology

基　　金：湖南省自然科学基金(No.2023JJ4019)。

摘　　要：目的探究过表达长链非编码RNA(long noncoding RNA,lncRNA)B3GALT5-AS1对非小细胞肺癌(non-small cell lung cancers,NSCLC)生长和侵袭能力的影响。方法qRT-PCR检测临床样本中B3GALT5-AS1的表达。A549细胞分对照组、pcDNA-scramble组、pcDNA-B3GALT5-AS1组,按分组分别转染pcDNA-scramble和pcDNA-B3GALT5 AS1载体。菌落形成实验评估A549细胞集落形成能力,qRT-PCR检测Ki67和Survivin的mRNA表达;流式细胞术分析细胞凋亡,Transwell检测细胞侵袭,蛋白质印迹检测P27、P53、VEGF和Vimentin蛋白表达。转染pcDNA-scramble和pcDNA-B3GALT5-AS1的A549细胞分别皮下注射裸鼠,检测瘤组织体积和重量;qRT-PCR检测B3GALT5-AS1的表达水平;免疫组化检测Ki67和VEGF的表达。结果肺癌组织中B3GALT5-AS1 mRNA相对表达量明显低于邻近正常组织(P<0.05)。与对照组比较,过表达B3GALT5-AS1 A549细胞集落形成能力降低(P<0.05),Survivin和Ki67表达量明显下调(P<0.05),细胞凋亡率明显升高(P<0.05),侵袭细胞数目明显减少(P<0.05),P27和P53蛋白表达显著上调(P<0.05),VEGF和Vimentin蛋白表达显著下调(P<0.05)。体内异种移植实验结果显示,与pcDNA-scramble组比较,过表达B3GALT5-AS1明显减小肿瘤体积和重量(P<0.05),瘤组织中Ki67和VEGF阳性细胞数目明显减少(P<0.05)。结论过表达B3GALT5-AS1明显抑制NSCLC细胞的增殖和侵袭并促进凋亡,而且体内阻滞瘤组织的生长,在NSCLC起到抗癌作用。Objective To explore the effect of overexpression of long non-coding RNA(lncRNA)B3GALT5-AS1 on the growth and invasion ability in non-small cell lung cancer(NSCLC).Methods qRT-PCR was employed to examine the expression level of B3GALT5-AS1 in clinical samples.A549 cells were divided into 3 groups:control group,pcDNA-scramble group,and pcDNA-B3GALT5-AS1 group,the pcDNA-scramble and pcDNA-B3GALT5 AS1 vectors were transfected into cells respectively.The colony formation experiment was employed to evaluate the colony-forming ability of A549 cells,qRT-PCR was used to detect the mRNA expression level of Ki67 and Survivin.Flow cytometry was used to analyse cell apoptosis,Transwell assay was used to test cell invasion,and Western blotting was used to detect the expression level of P27,P53,VEGF and Vimentin proteins.A549 cells transfected with pcDNA-scramble or pcDNA-B3GALT5-AS1 were injected subcutaneously into nude mice.The tumor volume and weight were measured,the expression level of B3GALT5-AS1 in tumor tissues was detected by qRT-PCR,and the expression of Ki67 and VEGF were determinded by immunohistochemistry.Results The relative mRNA expression level of B3GALT5-AS1 in the lung cancer tissues was significantly lower than that in the adjacent normal tissues when compared with that in the paracancerous tissues(P<0.05).The colony forming ability of cells in the pcDNA-B3GALT5-AS1 group was reduced(P<0.05),the expression of Ki67 and Survivin was obviously down-regulated(P<0.05),the apoptosis rate of cell was clearly increased(P<0.05),and the number of invasion cells was evidently reduced(P<0.05),the expression level of P27 and P53 proteins was up-regulated(P<0.05),and the expression of VEGF and Vimentin protein was significantly down-regulated(P<0.05).The results of in vivo xenotransplantation experiments showed that overexpression of B3GALT5-AS1 clearly reduced tumor volume and weight(P<0.05),and the numbers of Ki67 and VEGF positive cells in tumor tissues were obviously reduced(P<0.05)when compared with that in the pcDN

关 键 词：非小细胞肺癌 lncRNA B3GALT5-AS1 克隆形成 TRANSWELL 细胞凋亡 

分 类 号：R392.11[医药卫生—免疫学]