SNORD89 promotes stemness phenotype of ovarian cancer cells by regulating Notch1-c-Myc pathway
SNORD89 promotes stemness phenotype of ovarian cancer cells by regulating Notch1-c-Myc pathway
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SNORD89通过调节Notch1-c-Myc通路促进卵巢癌细胞的干细胞表型
DOI：
10.1186/s12967-019-2005-1
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发表时间：
2019-08-08
影响因子：
7.4
通讯作者：
Wei, Minjie
中科院分区：
文献类型：
--
作者：
Zhu, Wenjing;Niu, Jumin;Wei, Minjie
研究背景卵巢癌是妇科恶性肿瘤的主要死亡原因。肿瘤干细胞参与肿瘤的发生、发展和耐药。小核仁RNA（Small nucleolar RNA，SnoRNA）是一类小分子非编码RNA，与肿瘤细胞的干细胞性和肿瘤发生密切相关。方法本研究通过对TCGA数据库中379例卵巢癌患者的资料进行分析，筛选出与卵巢癌患者预后相关的SNORNA，并分析SNORNA在OVCAR-3（OV）sphere-forming（OS）细胞和OV细胞中表达的差异。过表达或敲除SNORD 89后，分别通过qRT-PCR或流式细胞术分析来测量OV、CAOV-3（CA）和OS细胞中Nanog、CD 44和CD 133的表达。采用CCK-8法、平板克隆形成法和软琼脂集落形成法检测细胞增殖和自我更新能力的变化。结果SNORD 89高表达提示卵巢癌患者预后不良，且与患者年龄、治疗效果有关。SNORD 89在卵巢癌干细胞中高表达。SNORD 89基因的过表达可使OV和CA细胞的干细胞标志物、S期细胞周期、细胞增殖、侵袭和迁移能力增加。相反，在OS细胞中SNORD 89沉默后，这些现象被逆转。进一步研究发现，SNORD 89可以上调c-Myc和Notch 1在mRNA和蛋白水平的表达。SNORD 89通过调控Notch 1-c-Myc通路促进细胞干细胞化，从而恶化卵巢癌患者的预后，是卵巢肿瘤发生的一个癌基因。因此，SNORD 89可能成为卵巢癌的新的预后生物标志物和治疗靶点。
BackgroundOvarian cancer is the leading cause of death in gynecological cancer. Cancer stem cells (CSCs) contribute to the occurrence, progression and resistance. Small nucleolar RNAs (SnoRNAs), a class of small molecule non-coding RNA, involve in the cancer cell stemness and tumorigenesis.MethodsIn this study, we screened out SNORNAs related to ovarian patient’s prognosis by analyzing the data of 379 cases of ovarian cancer patients in the TCGA database, and analyzed the difference of SNORNAs expression between OVCAR-3 (OV) sphere-forming (OS) cells and OV cells. After overexpression or knockdown SNORD89, the expression of Nanog, CD44, and CD133 was measured by qRT-PCR or flow cytometry analysis in OV, CAOV-3 (CA) and OS cells, respectively. CCK-8 assays, plate clone formation assay and soft agar colony formation assay were carried out to evaluate the changes of cell proliferation and self-renewal ability. Scratch migration assay and trans-well invasion analysis were used for assessing the changes of migration and invasion ability.ResultsHigh expression of SNORD89 indicates the poor prognosis of ovarian cancer patients and was associated with patients’ age, therapy outcome. SNORD89 highly expressed in ovarian cancer stem cells. The overexpression of SNORD89 resulted in the increased stemness markers, S phase cell cycle, cell proliferation, invasion and migration ability in OV and CA cells. Conversely, these phenomena were reversed after SNORD89 silencing in OS cells. Further, we found that SNORD89 could upregulate c-Myc and Notch1 expression in mRNA and protein levels. SNORD89 deteriorates the prognosis of ovarian cancer patients by regulating Notch1-c-Myc pathway to promote cell stemness and acts as an oncogene in ovarian tumorigenesis. Consequently, SNORD89 can be a novel prognostic biomarker and therapeutic target for ovarian cancer.