长链非编码RNA葡萄糖磷酸变位酶样蛋白5反义1通过靶向调控微小RNA-484对乳腺癌细胞的影响  

Effect of long-chain non-coding RNA phosphoglucomutase-like protein 5 antisense 1 on breast cancer cells by targeting microRNA-484

作　　者：杨天敬 唐瑞骏 刘海洋 韩璐 李然 李彩霞 Yang Tianjing;Tang Ruijun;Liu Haiyang;Han Lu;Li Ran;Li Caixia(Department of Breast and Thyroid Surgery,Tianjin Fourth Central Hospital,Tianjin 300140,China;Department of Pathology,Guilin Traditional Chinese Medicine Hospital,Guilin 541002,China;Department of Science and Education Section,Tianjin Fourth Central Hospital,Tianjin 300140,China;Department of Pathology,Tianjin Fourth Central Hospital,Tianjin 300140,China)

机构地区：[1]南开大学附属第四中心医院,天津医科大学第四中心临床学院,天津市第四中心医院乳腺甲状腺外科,300140 [2]桂林市中医医院病理科,541002 [3]南开大学附属第四中心医院,天津医科大学第四中心临床学院,天津市第四中心医院科教科,300140 [4]南开大学附属第四中心医院,天津医科大学第四中心临床学院,第四中心医院病理科,300140

出　　处：《中华实验外科杂志》2020年第9期1708-1711,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨长链非编码RNA(lncRNA)葡萄糖磷酸变位酶样蛋白5反义1(PGM5-AS1)靶向微小RNA(miR)-484对乳腺癌细胞增殖、细胞周期分布和凋亡的影响及其临床意义。方法53例乳腺癌患者来源于2016年1月至2018年10月南开大学附属第四中心医院确诊患者。收集患者肿瘤组织和癌旁组织(距离癌灶>5 cm处正常组织)。实时定量反转录聚合酶链反应(RT-qPCR)检测乳腺癌组织、癌旁组织、人乳腺上皮细胞株MCF-10A、乳腺癌细胞(MCF-7、MDA-MB-231、SKBR-3)中PGM5-AS1和miR-484的表达水平。将MCF-7细胞分为正常对照(NC)、pcDNA、pcDNA-PGM5-AS1、抗miR-con、抗miR-484、pcDNA-PGM5-AS1+miR-con、pcDNA-PGM5-AS1+miR-484组。采用噻唑蓝(MTT)实验、流式细胞术检测细胞的增殖活力、细胞凋亡率以及细胞周期分布。两组比较采用t检验;多组比较采用单因素方差分析,进一步两两比较采用SNK-q检验。结果与癌旁组织比较,乳腺癌组织PGM5-AS1表达(0.61±0.26比1.07±0.14)降低,miR-484表达(3.15±0.84比1.04±0.17)升高(t=11.341、17.924,P<0.05),差异有统计学意义。与MCF-10A细胞比较,乳腺癌细胞PGM5-AS1表达(0.28±0.03比1.04±0.07、0.37±0.05比1.04±0.07、0.35±0.04比1.04±0.07)降低,miR-484表达(4.75±0.34比1.07±0.12、3.89±0.48比1.07±0.12、3.58±0.42比1.07±0.12)升高(F=459.394、167.657,P<0.05),差异有统计学意义。与pcDNA组比较,pcDNA-PGM5-AS1组MCF-7细胞存活率[(68.28±4.78)%比(98.63±6.57)%]降低,凋亡率[(13.44±1.28)%比(4.63±0.59)%]、G0~G1期细胞比例[(72.42±3.64)%比(56.80±2.47)%]升高(F=89.925、363.156、81.190,P<0.05),差异有统计学意义。与抗miR-con组比较,抗miR-484组MCF-7细胞存活率[(64.64±7.26)%比(102.49±6.84)%]降低,凋亡率[(19.54±2.88)%比(4.17±0.57)%]、G0~G1期细胞比例[(68.38±2.60)%比(55.16±2.78)%]升高(F=97.967、253.890、59.086,P<0.05),差异有统计学意义。PGM5-AS1靶向负调控miR-484表达。结论PGM5-AS1通过靶向miR-484可抑制乳腺癌�Objective To investigate the effects of long-chain non-coding RNA(lncRNA)phosphoglucomutase-like protein 5 antisense 1(PGM5-AS1)targeting miR-484 on breast cancer cell proliferation,cell cycle distribution,and apoptosis and its clinical significance.Methods Fifty-three breast cancer patients were diagnosed in the Fourth Central Hospital affiliated to Nankai University from January 2016 to October 2018.Tumor tissue and paracancer tissue(normal tissue were more than 5cm away from neoplastci foci)were collected.Real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)was used to detect PGM5-AS1 and miR-484 expression levels in breast cancer tissues,adjacent tissues,human breast epithelial cell line MCF-10A,and breast cancer cells(MCF-7,MDA-MB-231,SKBR-3).MCF-7 cells were divided into normal control(NC),pcDNA,pcDNA-PGM5-AS1,anti-miR-con,anti-miR-484,pcDNA-PGM5-AS1+miR-con,pcDNA-PGM5-AS1+miR-484 groups.The cell proliferation,cell cycle and cell apoptosis in different groups were detected by methyl thiazolyl tetrazolium(MTT)and flow cytometry assays.The targeted and regulated relationship between PGM5-AS1 and miR-484 was confirmed by luciferase reporter assay and RT-qPCR.T-test was used for comparison between the two groups.One-way analysis of variance was used for multi-group comparison,and SNK-q test was used for further pair-wise comparison.Results Compared with adjacent tissues,the expression of PGM5-AS1 in breast cancer tissues(0.61±0.26 vs.1.07±0.14)decreased,and the expression of miR-484(3.15±0.84 vs.1.04±0.17)increased(t=11.341,17.924,P<0.05),the difference was statistically significant.Compared with MCF-10A cells,the expression of PGM5-AS1 in breast cancer cells(0.28±0.03 vs.1.04±0.07,0.37±0.05 vs.1.04±0.07,0.35±0.04 vs.1.04±0.07)decreased,miR-484 expression(4.75±0.34 vs.1.07±0.12,3.89±0.48 vs.1.07±0.12,3.58±0.42 vs.1.07±0.12)increased(F=459.394,167.657,P<0.05),the difference was statistically significant.Compared with the pcDNA group,the survival rate of MCF-7 cells in th

关 键 词：葡萄糖磷酸变位酶样蛋白5反义1 微小RNA-484 乳腺癌 细胞增殖 凋亡 细胞周期 

分 类 号：R737.9[医药卫生—肿瘤]