摘要
目的探讨PGM5反义RNA 1(PGM5-AS1)对食管癌细胞侵袭、迁移和Wnt信号通路的影响。方法采用实时荧光定量PCR(QPCR)检测人食管上皮细胞HET-1A和食管癌细胞(TE-1、KYSE150、KYSE450和EC-109)的PGM5-AS1水平;体外向EC-109细胞转染PGM5-AS1过表达载体(过表达组)或空载体pcDNA3.1(空载组),并设未转染的细胞为对照组。采用QPCR、MTT比色法、划痕实验和Transwell小室实验检测PGM5-AS1水平、增殖活力、划痕愈合率和穿膜细胞数,Western blotting检测Wnt1和β-连环蛋白(β-catenin)水平。结果食管癌细胞的PGM5-AS1水平低于HET-1A细胞(P<0.05);过表达组EC-109细胞的PGM5-AS1水平为19.772±1.980,高于对照组的1.067±0.059和空载组的0.923±0.087(P<0.05);过表达组转染48、72 h后的细胞增殖活力低于其余两组(P<0.05);过表达组的划痕愈合率和穿膜细胞数分别为(24.854±2.290)%和(60.539±26.577)个,低于对照组的(59.381±4.790)%和(165.421±22.082)个及空载组的(61.266±3.325)%和(157.758±22.462)个,差异有统计学意义(P<0.05);与对照组和空载组相比,过表达组的Wnt1和β-catenin水平降低(P<0.05)。对照组和空载组上述指标的差异无统计学意义(P>0.05)。结论食管癌细胞中PGM5-AS1表达降低并发挥抑癌基因的作用,上调PGM5-AS1水平可抑制增殖、迁移和侵袭能力,可能通过调控Wnt/β-catenin信号通路来发挥作用,有望成为食管癌治疗的潜在靶点。
Objective To investigate the effect of PGM5 antisense RNA 1(PGM5-AS1)on invasion,migration and Wnt signaling pathway of esophageal cancer cells.Methods Real time quantitative PCR(QPCR)was used to detect the PGM5-AS1 level in human esophageal epithelial cells HET-1A and esophageal cancer cells(TE-1,KYSE150,KYSE450 and EC-109).EC-109 cells were transfected with PGM5-AS1 overexpression vector(overexpression group)or empty vector pcDNA3.1(empty group)in vitro and cells without transfection were used as control group.QPCR,MTT colorimetry,scratch test and transwell chamber test were used to measure PGM5-AS1 level,proliferation activity,scratch healing rate and number of transmembrane cells.Western blotting was used to detect levels of Wnt1 andβ-catenin.Results The PGM5-AS1 level of esophageal cancer cells was lower than that of HET-1A cells(P<0.05).PGM5-AS1 level of overexpression group was 19.772±1.980,higher than 1.067±0.059 of control group and 0.923±0.087 of empty group(P<0.05).The proliferation activity of overexpression group at 48 and 72 h after transfection was lower than that of the other two groups(P<0.05).The scratch healing rate and number of transmembrane cells in overexpression group were(24.854±2.290)%and 60.539±26.577,lower than(59.381±4.790)%and 165.421±22.082 of control group and(61.266±3.325)%and 157.758±22.462 of empty group(P<0.05).Compared with control group and empty group,levels of Wnt1 andβ-catenin in overexpression group decreased(P<0.05).There was no significant difference in the above indexes between control group and empty group(P>0.05).Conclusion The decrease of PGM5-AS1 in esophageal cancer cells is accompanied by the role of tumor suppressor gene.Up-regulating PGM5-AS1 can inhibit the proliferation,migration and invasion.It may play a role by regulating the activity of Wnt/β-catenin signal pathway,which is expected to be a potential target of esophageal cancer treatment.
作者
梁云微
连相尧
党春艳
胡潺潺
朱翠敏
李爱科
李青山
LIANG Yunwei;LIAN Xiangyao;DANG Chunyan;HU Chanchan;ZHU Cuimin;LI Aike;LI Qingshan(Department of Oncology,the Affiliated Hospital of Chengde Medical College,Chengde 067000,China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2020年第1期7-12,共6页
Chinese Clinical Oncology
基金
承德市科技局科技支撑项目(20158002)
