Abstract：
We previously reported that long non-coding RNA (lncRNA) RPLP0P2 is involved in the progression of colorectal cancer (CRC); however, its molecular mechanisms in CRC remain unclear. In this study, we observed that RPLP0P2 was upregulated in CRC tissues and cell lines. Cell viability was measured using the MTT and colony formation assays. Migration and invasion capabilities were monitored by wound healing, transwell, and immunofluorescence assays. The results showed that RPLP0P2 downregulation inhibited cell viability, migration, and invasion capabilities of CRC cells, accompanied by decreased PCNA, N-cadherin, and Vimentin, and increased E-cadherin expression. Using the DIANA online database, miR-129-5p was identified as a downstream target of RPLP0P2. In fact, RPLP0P2 colocalized with miR-129-5p, acting as a miR-129-5p sponge. MiR-129-5p-inhibition almost abrogated the anti-tumor effects induced by RPLP0P2 inhibition in CRC cells. Zinc finger and BTB domain-containing 20 (ZBTB20) was identified as a potential downstream target of miR-129-5p in CRC cells. ZBTB20 overexpression prevented miR-129-5p mimic-mediated anti-tumor effects in CRC cells. A tumor xenograft assay was performed to monitor the role of RPLP0P2 in tumor growth. Of note, in tumor-bearing mice, RPLP0P2-silencing inhibited tumor growth, followed by increased miR-129-5p and decreased ZBTB20 expression. Our results suggest that lncRNA RPLP0P2 functions as an oncogene that promotes CRC cell proliferation and invasion via regulating the miR-129-5p/ZBTB20 axis, thus, it may serve as a candidate target for CRC interventional therapies.
摘要：
我们先前报道了长链非编码RNA(lncRNA)RPLP0P2参与结直肠癌(CRC)的进展;然而,其在CRC中的分子机制尚不清楚.在这项研究中,我们观察到RPLP0P2在CRC组织和细胞系中上调。使用MTT和集落形成测定法测量细胞活力。通过伤口愈合监测迁移和侵袭能力，transwell,和免疫荧光分析。结果表明，RPLP0P2下调抑制了细胞活力，迁移,以及CRC细胞的侵袭能力，伴随着PCNA的减少，N-钙黏着蛋白,还有Vimentin,E-cadherin表达增加。使用DIANA在线数据库，miR-129-5p被鉴定为RPLP0P2的下游靶标。事实上,RPLP0P2与miR-129-5p共定位，充当miR-129-5p海绵。MiR-129-5p抑制几乎消除了CRC细胞中RPLP0P2抑制诱导的抗肿瘤作用。锌指和含有BTB结构域的20(ZBTB20)被鉴定为CRC细胞中miR-129-5p的潜在下游靶标。ZBTB20过表达在CRC细胞中阻止miR-129-5p模拟物介导的抗肿瘤作用。进行肿瘤异种移植物测定以监测RPLP0P2在肿瘤生长中的作用。值得注意的是，在荷瘤小鼠中，RPLP0P2沉默抑制肿瘤生长，其次是miR-129-5p增加和ZBTB20表达减少。我们的结果表明，lncRNARPLP0P2作为癌基因，通过调节miR-129-5p/ZBTB20轴促进CRC细胞增殖和侵袭，因此，它可以作为CRC介入治疗的候选靶点.
