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01.12.2019 | Research | Ausgabe 1/2019 Open Access

Journal of Translational Medicine 1/2019

SNORD89 promotes stemness phenotype of ovarian cancer cells by regulating Notch1-c-Myc pathway

Zeitschrift:
Journal of Translational Medicine > Ausgabe 1/2019
Autoren:
Wenjing Zhu, Jumin Niu, Miao He, Liwen Zhang, Xuemei Lv, Fangxiao Liu, Longyang Jiang, Jing Zhang, Zhaojin Yu, Lin Zhao, Jia Bi, Yuanyuan Yan, Qian Wei, Hong Huo, Yue Fan, Yuzong Chen, Jian Ding, Minjie Wei
Wichtige Hinweise

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1186/​s12967-019-2005-1) contains supplementary material, which is available to authorized users.

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Abstract

Background

Ovarian 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.

Methods

In 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.

Results

High 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.
Zusatzmaterial
Additional file 5: Figure S4. Effects of SNORD89 interference on biological behaviors in CAOV-3 cells. a The SNORD89 expression in CA cells transfected with over expression (OE) of SNORD89 plasmid or negative control (NC) plasmid at 24, 48, and 72 h by qRT-PCR. The SNORD89 expression in CA cells transfected with NC plasmids was set as 1. b The mRNA expression of CD133, CD44 and Nanog were detected in CA cells transfected with SNORD89 OE or NC plasmids at 24 h by qRT-PCR. The mRNA expression of these genes in CA cells transfected with NC plasmids was set as 1. c The cell proliferation was measured in CA cells of SNORD89 overexpression by Cell Counting Kit-8 (CCK-8) assays at 24 h, 48 h, 72 h and 96 h transfection. d The cell proliferation ability was measured in CA cells of SNORD89 overexpression by plate clone formation assay. e The cell self-renewal ability was measured in CA cells of SNORD89 overexpression by soft agar colony formation assay. f The effect of SNORD89 overexpression on the migration ability of ovarian cancer cells by scratch migration assay in CA cells 24 h, 48 h and 72 h after transfection with SNORD89 OE plasmids. g The effect of SNORD89 overexpression on the migration ability of ovarian cancer cells by cell invasion analysis in CA cells 48 h after transfection with SNORD89 OE plasmids. h The mRNA expression of c-Myc and Notch1 was detected in CA cells transfected with SNORD89 OE or NC plasmids at 24 h by qRT-PCR. The mRNA expression of the two genes in CA cells transfected with NC plasmids was set as 1. i The representative western blot photos showed the increased expression of c-Myc and Notch1 in CA cells transfected with SNORD89 OE.
Literatur
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