Department of Gastrointestinal Surgery, Jiaozhou Central Hospital, Qingdao City, 266300, Shandong Province, China.
Department of General Surgery, Qingdao Municipal Hospital (Headquarters), No. 99 Yunxi Henan Road, Jiaozhou District, Shandong Province, 266000, Qingdao City, China.
In Vitro Cell Dev Biol Anim. 2022 Jun;58(6):462-470. doi: 10.1007/s11626-022-00686-7. Epub 2022 Jun 30.
The function of long non-coding RNA (lncRNA) RPL34-AS1 and microRNA (miR-3656) has been studied in several types of cancer, but their role in colorectal cancer (CRC) is unclear. We predicted that they could interact with each other; this study was carried out to explore their interaction in CRC. The expression of RPL34-AS1 and miR-3656 in CRC tissues and their paired non-tumor tissues from 62 CRC patients was determined by RT-qPCR. The direct interaction between RPL34-AS1 (both WT and mutant) and miR-3656 was determined by RNA-RNA pull-down assay. The interaction between them was studied with overexpression assay. Their role in cell proliferation was analyzed with BrdU assay. The role of RPL34-AS1 in regulating the expression of ACAP2 was explored by RT-qPCR and Western blot analysis. In this study, increased expression levels of miR-3656 and decreased expression levels of RPL34-AS1 were observed in CRC tissues. MiR-3656 directly interacted with RPL34-AS1, but not the RPL34-AS1 mutant with disrupted binding sites. RPL34-AS1 and miR-3565 did not affect the expression of each other. RPL34-AS1 suppressed the role of miR-3565 in enhancing cell proliferation, while RPL34-AS1 mutant did not affect cell behaviors and the role of miR-3565 in cell proliferation. RPL34-AS1 positively regulated the expression of ACAP2 at both mRNA and protein levels. Therefore, RPL34-AS1 is downregulated in CRC and may sponge miR-3656 to suppress cell proliferation in CRC.
长链非编码 RNA（lncRNA）RPL34-AS1 和 microRNA（miR-3656）的功能已在几种类型的癌症中得到研究，但它们在结直肠癌（CRC）中的作用尚不清楚。我们预测它们可以相互作用；本研究旨在探讨它们在 CRC 中的相互作用。通过 RT-qPCR 测定了 62 例 CRC 患者的 CRC 组织及其配对非肿瘤组织中 RPL34-AS1 和 miR-3656 的表达。通过 RNA-RNA 下拉测定确定 RPL34-AS1（WT 和突变体）和 miR-3656 之间的直接相互作用。通过过表达测定研究它们之间的相互作用。通过 BrdU 测定分析它们在细胞增殖中的作用。通过 RT-qPCR 和 Western blot 分析探索 RPL34-AS1 在调节 ACAP2 表达中的作用。在这项研究中，观察到 CRC 组织中 miR-3656 的表达水平升高，而 RPL34-AS1 的表达水平降低。miR-3656 与 RPL34-AS1 直接相互作用，但与结合位点破坏的 RPL34-AS1 突变体不相互作用。RPL34-AS1 和 miR-3565 不影响彼此的表达。RPL34-AS1 抑制了 miR-3565 增强细胞增殖的作用，而 RPL34-AS1 突变体不影响细胞行为和 miR-3565 在细胞增殖中的作用。RPL34-AS1 正向调节 ACAP2 在 mRNA 和蛋白水平的表达。因此，RPL34-AS1 在 CRC 中下调，可能通过海绵吸附 miR-3656 抑制 CRC 中的细胞增殖。
