环状 WHSC1 通过海绵吸附 miR-145 和 miR-1182 调控 MUC1 和 hTERT 促进卵巢癌进展。

CircWHSC1 promotes ovarian cancer progression by regulating MUC1 and hTERT through sponging miR-145 and miR-1182.

机构信息

Department of Gynecologic Oncology Research Office, The Third Affiliated Hospital of Guangzhou Medical University, No.63 Duobao Road, Liwan District, Guangzhou, 510150, Guangdong, China.

Department of Obstetrics and Gynecology, Key Laboratory for Major Obstetric Diseases of Guangdong Province, and Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institute in Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510150, China.

出版信息

J Exp Clin Cancer Res. 2019 Oct 30;38(1):437. doi: 10.1186/s13046-019-1437-z.

DOI:10.1186/s13046-019-1437-z

PMID:31666098

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6822369/

Abstract

BACKGROUND

Circular RNAs are key regulators in human cancers, however, there is a lack of studies on circRNAs' specific functions in ovarian cancer.

METHODS

Our study used qRT-PCR to detect the differentially expressed circRNAs between normal ovaries and ovarian cancer tissues. Cell function experiments were performed to verify the role of overexpression and silence of circWHSC1, including MTT assay, cell apoptosis assay, wound healing and Matrigel-coated Transwell assay. In vivo tumorigenesis model was constructed by subcutaneous injection in nude mice. Bioinformatics analysis predicted the possible binding sites of circWHSC1 with miRNAs, and confirmed with dual-luciferase reporter assay and RNA pull-down assay. The exosomes were extracted with ultracentrifugation. HE staining was also used to detect morphology of nude mice peritoneum.

RESULTS

We found that circWHSC1 was up-regulated in ovarian cancer tissues, and circWHSC1 expression was higher in moderate & poor differentiation ovarian cancer tissues than in well differentiation ovarian cancer tissues. Overexpression of circWHSC1 increased cell proliferation, migration and invasion, and inhibited cell apoptosis. Silence of circWHSC1 exerted the opposite effects. Additionally, circWHSC1 could sponge miR-145 and miR-1182 and up-regulate the expression of downstream targets MUC1 and hTERT. Exosomal circWHSC1 can be transferred to peritoneal mesothelial cells and promotes peritoneal dissemination.

CONCLUSIONS

Our study demonstrates the highly expressed circWHSC1 in ovarian cancer promotes tumorigenesis by sponging miR-145 and miR-1182, and its exosome forms induce tumor metastasis through acting on peritoneal mesothelium.

摘要

背景

环状 RNA 是人类癌症中的关键调节剂，然而，关于环状 RNA 在卵巢癌中的特定功能的研究还很缺乏。

方法

我们的研究使用 qRT-PCR 检测正常卵巢组织和卵巢癌组织中差异表达的环状 RNA。通过 MTT 检测、细胞凋亡检测、划痕愈合和 Matrigel 包被 Transwell 检测等细胞功能实验，验证 circWHSC1 过表达和沉默的作用。通过裸鼠皮下注射构建体内肿瘤发生模型。生物信息学分析预测 circWHSC1 与 miRNAs 可能的结合位点，并通过双荧光素酶报告基因检测和 RNA 下拉实验进行验证。采用超速离心法提取外泌体。HE 染色也用于检测裸鼠腹膜的形态。

结果

我们发现 circWHSC1 在卵巢癌组织中上调，中-低分化卵巢癌组织中 circWHSC1 的表达高于高分化卵巢癌组织。circWHSC1 的过表达增加了细胞增殖、迁移和侵袭，并抑制了细胞凋亡。circWHSC1 的沉默则产生相反的效果。此外，circWHSC1 可以海绵吸附 miR-145 和 miR-1182，并上调下游靶标 MUC1 和 hTERT 的表达。外泌体 circWHSC1 可转移至腹膜间皮细胞，并促进腹膜扩散。