摘要
目的膀胱癌是目前人类十大常见恶性肿瘤之一,近20年来膀胱癌的发病率逐渐增加,且进展期患者5年生存率明显降低(≤30%).本研究旨在探讨转移相关肺腺癌转录本1(metastasis associated lung adenocarcinoma transcript 1,MALAT1)在膀胱癌细胞中的表达水平及其作用机制,以及长链非编码MALAT1对膀胱癌细胞迁移和侵袭行为的影响.方法qPCR检测MALAT1和miR-205在膀胱癌组织和不同膀胱癌细胞株中表达,双荧光素酶实验验证MAL-AT1对miR-205调控作用,划痕愈合实验和Transwell侵袭实验检测MALAT1和miR-205对膀胱癌细胞迁移和侵袭能力影响,蛋白质印迹法检测抑制MALAT1表达后JAK/STAT2信号通路蛋白表达.结果与癌旁正常膀胱癌组织(0.15±0.04)相比,MALAT1在膀胱癌组织表达增高(1.78±0.19),差异有统计学意义,t=12.26,P<0.05;并且MALAT1表达与膀胱癌的分级有关联,χ^2=5.973,P=0.015.双荧光素酶实验证实,MALAT1可以调控miR-205表达.抑制MALAT1表达后可以抑制膀胱癌细胞的迁移和侵袭能力,24、48、72 h细胞的迁移率分别为(8.15±1.59)%、(21.65±2.35)%、(61.08±5.16)%,P<0.05,侵袭细胞数量为58.62±7.16,P<0.05,差异有统计学意义;同时抑制miR-205表达水平后,膀胱癌细胞的迁移和侵袭能力得到一定程度的恢复,24、48、72 h细胞迁移率分别为(10.21±1.42)%、(25.35±4.32)%和(69.15±5.72)%,侵袭细胞数量为(476.19±28.26);抑制MALAT1表达后,JAK/STAT2蛋白表达受到抑制,同时抑制miR-205后JAK/STAT2表达水平相应上调.结论MALAT1抑制miR-205表达后通过JAK/STAT2信号通路影响膀胱癌细胞的迁移和侵袭能力.
OBJECTIVE Bladder cancer is one of the ten most common cancers in human malignant tumors.The incidence of bladder cancer is gradually increasing in the past 20 years,and the 5-year survival rate of patients in advanced stage is significantly lower than 30%.This study aimed to investigate the expression level of MALAT1 in bladder cancer cells and its mechanism of action,to determine the role of MALAT1 in bladder cancer,and explore the effect of long-chain non-coding MALAT1 on the migration and invasion of bladder cancer cells and its possible mechanism.METHODS qPCR was used to detect the expression of MALAT1 and miR-205 in different bladder cancer cell lines.To analyze the association between MALAT1 and clinicopathological data in patients with bladder cancer,double luciferase reporter gene was used to detect the interaction between MALAT1 and miR-205.Scaling healing experiment and transwell invasion assay were used to detect the effect of MALAT1 and miR-205 on migration and invasion of bladder cancer cells.Western blotting was used to detect the protein expression of JAK/STAT2 signaling pathway after inhibiting MALAT1 expression.RESUITS Compared with adjacent normal bladder cancer tissues,MALAT1 was increased in bladder cancer tissues[(0.15±0.04)vs(1.78±0.19),t=12.26,P<0.05],and the expression of MALAT1 was positively correlated with the staging of bladder cancer.Double luciferase assay confirmed that MALAT1 could specifically bind to the 3′UTR of miR-205 and regulate the expression and activity of miR-205.Inhibiting the MALAT1 expression could inhibit the migration[24 h(8.15±1.59)%,48 h(21.65±2.35)%,72 h(61.08±5.16)%,P<0.05]and invasion[(58.62±7.16),P<0.05]of bladder cancer cells.While the inhibiting the miR-205 expression level,the migration[24 h(10.21±1.42)%,48 h(25.35±4.32)%,72 h(69.15±5.72)%,P<0.05]and invasion [(476.19±28.26),P<0.05]ability of bladder cancer cells were improved.After inhibiting the MALAT1 expression,the JAK/STAT2 signaling pathway was activated accordingly,and the expression of
作者
王哲
张敬
陈怀安
张潮
刘硕
苗文隆
WANG Zhe;ZHANG Jing;CHEN Huai-an;ZHANG Chao;LIU Shuo;MIAO Wen-long(Department of Urology,First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,P.R.China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2019年第18期1342-1347,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
河北省医学科学研究重点课题计划(20150472)
