MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma.
MiR-20a-5p suppresses tumor proliferation by targeting autophagy-related gene 7 in neuroblastoma.
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MiR-20a-5p 通过靶向神经母细胞瘤中自噬相关基因 7 抑制肿瘤增殖
DOI：
10.1186/s12935-017-0499-2
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发表时间：
2018
影响因子：
5.8
通讯作者：
Ni X
中科院分区：
文献类型：
--
作者：
Yu Y;Zhang J;Jin Y;Yang Y;Shi J;Chen F;Han S;Chu P;Lu J;Wang H;Guo Y;Ni X
神经母细胞瘤（neuroblastoma， NB）是儿童最常见的恶性肿瘤，起源于颅外交感神经系统。这种疾病背后的分子机制很复杂，尚未完全了解。方法采用实时荧光定量PCR （quantitative real-time PCR, qRT-PCR）技术，定量检测miR-20a-5p及其靶基因ATG7在临床NB组织中的表达。通过体外研究（Real-Time细胞动力学分析仪、集落形成实验、caspase-Glo 3/7实验和western blotting）研究miR-20a-5p和ATG7在SH-SY5Y细胞中的生物学功能。荧光素酶报告基因检测验证miR-20a-5p与ATG7之间的生物学关系。结果我们发现，miR-20a-5p的表达随着NB的临床分期而显著下调，而其靶自噬相关基因7 （ATG7）的表达则随着NB的进展而升高。相关分析显示，miR-20a-5p与ATG7呈负相关趋势。在SH-SY5Y细胞中，强迫表达miR-20a-5p抑制ATG7的表达、自噬启动和细胞增殖，同时促进细胞凋亡，提示miR-20a-5p与ATG7之间可能存在关联。进一步的生物信息学靶标预测结合蛋白表达和荧光素酶报告基因检测验证了miR-20a-5p通过直接结合ATG7的3′-UTR抑制ATG7，证实了miR-20a-5p参与NB中ATG7的调控。结论miR-20a-5p通过负调控ATG7抑制SH-SY5Y细胞自噬，从而抑制细胞增殖，促进细胞凋亡。因此，明确自噬在NB中的特定作用及其调控机制对于开发针对NB的自噬靶向治疗方法至关重要。miR-20a-5p和ATG7都将是未来NB治疗的潜在治疗靶点。
BackgroundNeuroblastoma (NB) is the most common malignant tumor originating from the extracranial sympathetic nervous system in children. The molecular mechanisms underlying this disease are complex, and not completely understood.MethodsQuantitative real-time PCR (qRT-PCR) was applied to quantify the expression of miR-20a-5p and its target gene ATG7 in clinical NB tissues. The biological function of miR-20a-5p and ATG7 in SH-SY5Y cells was investigated through in vitro studies (Real-Time cell kinetic analyzer, colony formation assay, caspase-Glo 3/7 assay and western blotting). The luciferase reporter assay was conducted to verify the biological relationship between miR-20a-5p and ATG7.ResultsHere we found that miR-20a-5p expression was significantly downregulated whereas its target autophagy-related gene 7 (ATG7) was increased along with clinical staging of NB progression. Correlation analysis showed that miR-20a-5p had a negative correlation trend with ATG7. In SH-SY5Y cells, forced expression of miR-20a-5p suppressed ATG7 expression, autophagy initiation and cellular proliferation while promoted apoptosis, suggesting a potential association between miR-20a-5p and ATG7. Further bioinformatic target prediction combined with protein expression and luciferase reporter assay verified that miR-20a-5p inhibited ATG7 by directly binding to its 3′-UTR, confirming the involvement of miR-20a-5p in the regulation of ATG7 in NB.ConclusionsThese results clarified that miR-20a-5p inhibited cell proliferation and promoted apoptosis through negative regulation of ATG7 and thus autophagy suppression in SH-SY5Y cells. Therefore, defining the context-specific roles of autophagy in NB and regulatory mechanisms involved will be critical for developing autophagy-targeted therapeutics against NB. Both miR-20a-5p and ATG7 would be potential therapeutic targets for future NB treatment.
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DOI：
10.1158/1078-0432.ccr-11-3321
发表时间：
2012-07-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Li Z;Yan S;Attayan N;Ramalingam S;Thiele CJ
通讯作者：
Thiele CJ
DOI：
10.3322/caac.21244
发表时间：
2014-09
期刊：
CA: a cancer journal for clinicians
影响因子：
--
作者：
Berindan-Neagoe I;Monroig Pdel C;Pasculli B;Calin GA
通讯作者：
Calin GA
影响因子：
51.1
作者：
Oberg, Kjell;Modlin, Irvin M.;De Herder, Wouter;Pavel, Marianne;Klimstra, David;Frilling, Andrea;Metz, David C.;Heaney, Anthony;Kwekkeboom, Dik;Strosberg, Jonathan;Meyer, Timothy;Moss, Steven F.;Washington, Kay;Wolin, Edward;Liu, Eric;Goldenring, James
通讯作者：
Goldenring, James
癌症中的microRNA生物发生途径。
DOI：
10.1038/nrc3932
发表时间：
2015-06
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
Lin S;Gregory RI
通讯作者：
Gregory RI
自噬与神经母细胞瘤的化学抗性有关。
影响因子：
3.8
作者：
Belounis A;Nyalendo C;Le Gall R;Imbriglio TV;Mahma M;Teira P;Beaunoyer M;Cournoyer S;Haddad E;Vassal G;Sartelet H
通讯作者：
Sartelet H