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Synthetic tetracycline-controllable shRNA targeting long non-coding RNA HOXD-AS1 inhibits the progression of bladder cancer
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نویسنده
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li j. ,zhuang c. ,liu y. ,chen m. ,chen y. ,chen z. ,he a. ,lin j. ,zhan y. ,liu l. ,xu w. ,zhao g. ,guo y. ,wu h. ,cai z. ,huang w.
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منبع
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journal of experimental and clinical cancer research - 2016 - دوره : 35 - شماره : 1
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چکیده
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Background: long non-coding rnas (lncrnas) have been proved to act as key molecules in cancer development and progression. dysregulation of lncrnas is discovered in various tumor tissues and cancer cells where they can serve as oncogenes or tumor suppressors. long non-coding rna hoxd-as (hoxd cluster antisense rna 1) has recently been identified to be involved in the development of several cancers including neuroblastoma,adenocarcinomas and breast cancer. however,the role of hoxd-as1 in bladder cancer remains unknown. methods: the synthetic tetracycline-controllable shrna was used to modulate the level of hoxd-as1 by adding different concentrations of doxycycline (dox). rt-qpcr was used to detect the expression level of hoxd-as1. cell proliferation was determined by cck-8 assay and edu incorporation experiment when hoxd-as1 was knocked down. we used wound-healing assay for detecting the effect of hoxd-as1 on cell migration. eventually,cell apoptosis was determined by caspase 3 elisa assay and flow cytometry assay. results: in this study,we found that the expression level of hoxd-as1 was significantly increased in bladder cancer tissues and cells. furthermore,high expression of hoxd-as1 was significantly related to tumor size,histological grade and tnm stage. in vitro assays confirmed that knockdown of hoxd-as1 suppressed cell proliferation/migration and increased the rate of apoptotic cell in bladder cancer cells. at last,we used the important element of synthetic biology,tetracycline(tet)-controllable switch,to construct tet-controllable shrna vectors which can modulate the expression of hoxd-as1 in a dosage-dependent manner. conclusions: our research suggested that high expression of hoxd-as1 may be involved in the bladder cancer carcinogenesis through inhibiting the phenotypes and activating endogenous cancer-related molecular pathways. therefore,hoxd-as1 may act as an oncogene and provide a potential attractive therapeutic target for bladder cancer. in addition,the synthetic tetracycline-controllable shrna may provide a novel method for cancer research in vitro assays. © 2016 the author(s).
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کلیدواژه
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Bladder cancer; HOXD-AS1; lncRNA; Synthetic biology; Tetracycline-controllable shRNA
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آدرس
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key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041,china,guangdong and shenzhen key laboratory of male reproductive medicine and genetics,institute of urology,peking university shenzhen hospital,shenzhen pku-hkust medical center,shenzhen,518036, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,anhui medical university,hefei,anhui province,230000, China, shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,anhui medical university,hefei,anhui province,230000, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039, China, shanghai-most key laboratory of health and disease genomics,chinese national human genome centerat shanghai,shanghai,200000, China, department of urology,peking university first hospital,institute of urology,peking university,national urological cancer center,beijing,100034, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041,china,anhui medical university,hefei,anhui province,230000,china,department of urology,peking university first hospital,institute of urology,peking university,national urological cancer center,beijing,100034, China, key laboratory of medical reprogramming technology,shenzhen second people's hospital,clinical institute of shantou,university medical college,first affiliated hospital,shenzhen university,shenzhen,guangdong province,518039,china,shantou university medical college,shantou,guangdong province,515041,china,anhui medical university,hefei,anhui province,230000,china,department of urology,peking university first hospital,institute of urology,peking university,national urological cancer center,beijing,100034, China
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