發(fā)布時(shí)間：2018-09-08 11:54

【摘要】：目的:探討miRNA-20a在膀胱癌組織中的表達(dá)及其機(jī)制。方法:收集2014年1月至2015年1月昆明醫(yī)科大學(xué)第二附屬醫(yī)院96例患者的組織標(biāo)本,運(yùn)用實(shí)時(shí)定量聚合酶鏈反應(yīng)(q RT-PCR)方法檢測(cè)miRNA-20a在膀胱癌組織和癌旁組織中的表達(dá);生物信息學(xué)方法預(yù)測(cè)miRNA-20a的靶基因并采用雙熒光素酶報(bào)告基因?qū)嶒?yàn)進(jìn)行驗(yàn)證;q RT-PCR、Western blot以及細(xì)胞免疫熒光分別檢測(cè)人膀胱癌細(xì)胞系T24和J82細(xì)胞轉(zhuǎn)染miRNA-20a模擬物或陰性對(duì)照后對(duì)靶基因mRNA和蛋白表達(dá)的影響;CCK-8、Transwell侵襲小室和劃痕實(shí)驗(yàn)檢測(cè)過(guò)表達(dá)miRNA-20a后T24細(xì)胞體外增殖、遷移和侵襲能力的改變。結(jié)果:miRNA-20a在膀胱癌組織中高表達(dá),且與腫瘤的病理分級(jí)、臨床分期、轉(zhuǎn)移以及復(fù)發(fā)密切相關(guān)(P0.001);雙熒光素酶報(bào)告基因?qū)嶒?yàn)證實(shí)miRNA-20a與人源性長(zhǎng)壽保障基因2(Homo sapiens longevity assurance homologue 2,LASS2)的3'-UTR直接靶向結(jié)合;轉(zhuǎn)染miRNA-20a模擬物可顯著下調(diào)膀胱癌細(xì)胞中LASS2 m RNA和蛋白的表達(dá)(P0.01),增強(qiáng)膀胱癌細(xì)胞的增殖、侵襲和遷移能力(P0.01)。結(jié)論:miRNA-20a在膀胱癌組織中高表達(dá),且miRNA-20a可通過(guò)靶向抑制LASS2促進(jìn)膀胱癌細(xì)胞的增殖、侵襲和遷移,與膀胱癌的發(fā)生發(fā)展相關(guān)。
[Abstract]:Objective: to investigate the expression and mechanism of miRNA-20a in bladder cancer. Methods: from January 2014 to January 2015, 96 tissue specimens from the second affiliated Hospital of Kunming Medical University were collected and the expression of miRNA-20a in bladder cancer and adjacent tissues was detected by real-time quantitative polymerase chain reaction (Q RT-PCR). Bioinformatics method was used to predict the target gene of miRNA-20a, and double luciferase reporter gene assay was used to verify the expression of miRNA-20a mimics or negative control in human bladder cancer cell lines T24 and J82, respectively, by using double luciferase reporter gene assay and immunofluorescence assay. The expression of target gene mRNA and protein was affected by CCK-8 transwell invasion chamber and scratch assay was used to detect the proliferation of T24 cells after overexpression of miRNA-20a in vitro. Changes in migration and invasiveness. Results the expression of 10 miRNA-20a was highly expressed in bladder cancer tissues, and was correlated with the pathological grade and clinical stage of the tumor. There was a close correlation between metastasis and recurrence (P0.001), double luciferase reporter gene experiment confirmed that miRNA-20a was directly targeted to the 3'-UTR of human longevity guarantee gene 2 (Homo sapiens longevity assurance homologue 2 / LASS2. Transfection of miRNA-20a mimics could significantly down-regulate the expression of LASS2 m RNA and protein in bladder cancer cells (P0.01), and enhance the proliferation, invasion and migration of bladder cancer cells (P0.01). ConclusionMmiRNA-20a is highly expressed in bladder cancer tissues, and miRNA-20a can promote the proliferation, invasion and migration of bladder cancer cells by targeting the inhibition of LASS2, which is related to the occurrence and development of bladder cancer.
【作者單位】： 昆明醫(yī)科大學(xué)第二附屬醫(yī)院泌尿外科云南省泌尿外科研究所;云南省楚雄州人民醫(yī)院新區(qū)腎臟內(nèi)科;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(編號(hào):81460384) 云南省科技計(jì)劃項(xiàng)目(編號(hào):2015FB196) 云南省教育廳科學(xué)研究基金項(xiàng)目(編號(hào):2014Z072)資助~~
【分類(lèi)號(hào)】：R737.14

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