The long non-coding RNA BC200 (BCYRN1) is critical for cancer cell survival and proliferation.
The long non-coding RNA BC200 (BCYRN1) is critical for cancer cell survival and proliferation.
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DOI：
10.1186/s12943-017-0679-7
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发表时间：
2017-06-26
期刊：
影响因子：
37.3
通讯作者：
McKenna SA
中科院分区：
文献类型：
--
作者：
Booy EP;McRae EK;Koul A;Lin F;McKenna SA
BC 200是一种长的非编码RNA，在大脑中以高水平表达，并在多种肿瘤类型中升高。BC 200在翻译调节中具有假设的作用;然而，迄今为止，BC 200在正常和疾病状态中的功能作用仍然很差。通过定量实时PCR在肿瘤细胞系、原代和非致瘤性培养的乳腺和肺细胞以及一组正常人组织中进行详细的BC 200表达分析，并通过北方印迹证实。进行亚细胞分级分离以评估BC 200分布，并使用锁核酸（LNA）GapmeR和常规siRNA两者建立BC 200的有效敲低。MTT法检测BC 200基因敲减和过表达后细胞活力，Annexin V/PI染色和流式细胞术检测细胞凋亡。细胞周期阻滞和同步化使用血清提取以及特异性抑制剂洛伐他汀、胸苷、RO 3306和诺考达唑进行。通过流式细胞术对细胞DNA含量的荧光分析监测同步化，在脑中BC 200表达显著上调，并且在睾丸、小肠和卵巢中也观察到表达升高。在培养的肿瘤细胞中的表达显着高于相应的正常组织，然而，在培养的原代细胞中的表达是类似的永生化和癌细胞系。BC 200敲低导致通过生长停滞和诱导细胞凋亡的活力的急剧丧失，这可以部分地通过野生型BC 200的过表达而不是siRNA抗性序列突变体来挽救。在细胞汇合或血清剥夺后观察到BC 200表达的大幅减少，以及药物诱导的细胞周期停滞在G1或G2期，但不是S期或M期。在从细胞周期停滞释放后，BC 200表达随着细胞进入S期而恢复，但在细胞周期的同步进展期间不遵循周期性表达模式。这种升高的表达对于增殖的癌细胞和非癌细胞的存活是至关重要的，但在衰老或细胞周期停滞时是不稳定的。BC 200表达在增殖培养细胞中升高，而与来源无关。在原代细胞中，表达在通过汇合、血清剥夺或化学抑制的细胞周期停滞时显著降低。BC 200敲低的致死性仅限于活跃增殖的细胞，使其成为广谱癌症的有希望的治疗靶点。本文的在线版本（doi：10.1186/s12943-017-0679-7）包含补充材料，可供授权用户使用。
BC200 is a long non-coding RNA expressed at high levels in the brain and elevated in a variety of tumour types. BC200 has a hypothesized role in translational regulation; however, to date the functional role of BC200 in both normal and diseased states remains poorly characterized. Detailed BC200 expression analyses were performed in tumor cell lines, primary and non-tumorigenic cultured breast and lung cells, and a panel of normal human tissues by quantitative real-time PCR and confirmed by northern blot. Subcellular fractionation was performed to assess BC200 distribution and efficient knock-down of BC200 was established using both locked nucleic acid (LNA) GapmeRs and conventional siRNAs. Cell viability following BC200 knockdown and overexpression was assessed by MTT assay and induction of apoptosis was monitored by Annexin V/PI staining and flow cytometry. Cell cycle arrest and synchronization were performed using serum withdrawal as well as the specific inhibitors Lovastatin, Thymidine, RO3306 and Nocodazole. Synchronization was monitored by fluorescent analysis of cellular DNA content by flow cytometry BC200 expression was substantially upregulated in brain and elevated expression was also observed in testes, small intestine and ovary. Expression in cultured tumour cells was dramatically higher than corresponding normal tissue; however, expression in cultured primary cells was similar to that in immortalized and cancer cell lines. BC200 knockdown resulted in a dramatic loss of viability through growth arrest and induction of apoptosis that could be partially rescued by overexpression of wild-type BC200 but not an siRNA-resistant sequence mutant. A substantial decrease in BC200 expression was observed upon cell confluence or serum deprivation, as well as drug induced cell cycle arrest in G1 or G2 but not S- or M-phases. Upon release from cell cycle arrest, BC200 expression was recovered as cells entered S-phase, but did not follow a periodic expression pattern during synchronized progression through the cell cycle. This elevated expression was critical for the survival of proliferating cancerous and non-cancerous cells, but is dispensable upon senescence or cell cycle arrest. BC200 expression is elevated in proliferating cultured cells regardless of origin. In primary cells, expression is dramatically reduced upon cell cycle arrest by confluence, serum deprivation or chemical inhibition. The lethality of BC200 knockdown is restricted to actively proliferating cells, making it a promising therapeutic target for a broad spectrum of cancers. The online version of this article (doi:10.1186/s12943-017-0679-7) contains supplementary material, which is available to authorized users.