Circular RNA ZNF800 (hsa_circ_0082096) regulates cancer stem cell properties and tumor growth in colorectal cancer

被引：9

作者：

Rengganaten, Vimalan ^{[1
,2
,3
]}

Huang, Chiu-Jung ^{[4
]}

Wang, Mong-Lien ^{[3
,5
,6
,7
]}

Chien, Yueh ^{[6
]}

Tsai, Ping-Hsing ^{[6
]}

Lan, Yuan-Tzu ^{[5
,8
]}

Ong, Hooi Tin ^{[9
,10
]}

Chiou, Shih-Hwa ^{[3
,5
,6
]}

Choo, Kong Bung ^{[1
,6
,10
]}

机构：

[1] Univ Tunku Abdul Rahman, Ctr Stem Cell Res, Kajang 43000, Selangor, Malaysia

[2] Univ Tunku Abdul Rahman, M Kandiah Fac Med & Hlth Sci, Postgrad Program, Kajang 43000, Malaysia

[3] Natl Yang Ming Chiao Tung Univ, Inst Pharmacol, Taipei 11221, Taiwan

[4] Chinese Culture Univ, Dept Anim Sci & Grad Inst Biotechnol, Taipei 11221, Taiwan

[5] Natl Yang Ming Chiao Tung Univ, Sch Med, Taipei 11221, Taiwan

[6] Taipei Vet Gen Hosp, Dept Med Res, Taipei 11221, Taiwan

[7] Natl Yang Ming Chiao Tung Univ, Inst Food Safety & Hlth Risk Assessment, Taipei 11221, Taiwan

[8] Taipei Vet Gen Hosp, Dept Surg, Div Colon & Rectal Surg, Taipei 11221, Taiwan

[9] Univ Tunku Abdul Rahman, Ctr Canc Res, Kajang 43000, Selangor, Malaysia

[10] Univ Tunku Abdul Rahman, M Kandiah Fac Med & Hlth Sci, Dept Preclin Sci, Kajang 43000, Selangor, Malaysia

来源：

BMC CANCER | 2023年 / 23卷 / 01期

关键词：

circZNF800-miRNA-mRNA regulation; Cancer stem cells; Colorectal cancer; Tumorigenesis; CRISPR Cas13d knockdown; METASTASIS; EXPRESSION; BLADDER; ORIGIN; NANOG;

D O I：

10.1186/s12885-023-11571-1

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

BackgroundCancer stem cells form a rare cell population in tumors that contributes to metastasis, recurrence and chemoresistance in cancer patients. Circular RNAs (circRNAs) are post-transcriptional regulators of gene expression that sponge targeted microRNA (miRNAs) to affect a multitude of downstream cellular processes. We previously showed in an expression profiling study that circZNF800 (hsa_circ_0082096) was up-regulated in cancer stem cell-enriched spheroids derived from colorectal cancer (CRC) cell lines.MethodsSpheroids were generated in suspension spheroidal culture. The ZNF800 mRNA, pluripotency stem cell markers and circZNF800 levels were determined by quantitative RT-PCR. CircZNF800-miRNA interactions were shown in RNA pulldown assays and the miRNA levels determined by stem-loop qRT-PCR. The effects of circZNF800 on cell proliferation were tested by EdU staining followed by flowcytometry. Expression of stem cell markers CD44/CD133, Lgr5 and SOX9 was demonstrated in immunofluorescence microscopy. To manipulate the cellular levels of circZNF800, circZNF800 over-expression was achieved via transfection of in vitro synthesized and circularized circZNF800, and knockdown attained using a CRISPR-Cas13d-circZNF800 vector system. Xenografted nude mice were used to demonstrate effects of circZNF800 over-expression and knockdown on tumor growth in vivo.ResultsCircZNF800 was shown to be over-expressed in late-stage tumor tissues of CRC patients. Data showed that circZNF800 impeded expression of miR-140-3p, miR-382-5p and miR-579-3p while promoted the mRNA levels of ALK/ACVR1C, FZD3 and WNT5A targeted by the miRNAs, as supported by alignments of seed sequences between the circZNF800-miRNA, and miRNA-mRNA paired interactions. Analysis in CRC cells and biopsied tissues showed that circZNF800 positively regulated the expression of intestinal stem cell, pluripotency and cancer stem cell markers, and promoted CRC cell proliferation, spheroid and colony formation in vitro, all of which are cancer stem cell properties. In xenografted mice, circZNF800 over-expression promoted tumor growth, while circZNF800 knockdown via administration of CRISPR Cas13d-circZNF800 viral particles at the CRC tumor sites impeded tumor growth.ConclusionsCircZNF800 is an oncogenic factor that regulate cancer stem cell properties to lead colorectal tumorigenesis, and may be used as a predictive marker for tumor progression and the CRISPR Cas13d-circZNF800 knockdown strategy for therapeutic intervention of colorectal cancer.

引用

页数：15