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. 2021 Jan 6;14:1–13. doi: 10.2147/OTT.S280912

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© 2021 He et al.

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Long noncoding RNA DCST1 antisense RNA 1 (CCDC144NL-AS1) acts as a molecular sponge for miR-490-3p in osteosarcoma (OS) cells. (A and B) lncRNA subcellular localization predictors lncATLAS and lncLocator were used to predict CCDC144NL-AS1 localization. (C) Subcellular fractionation assay showed that CCDC144NL-AS1 was mostly located in the cytoplasm of HOS and Saos-2 cells. (D) StarBase 3.0 and miRcode were used to identify the putative miRNAs targeting CCDC144NL-AS1. (E) Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to measure the mRNA expression in CCDC144NL-AS1-depleted HOS and Saos-2 cells. (F) RT-qPCR analysis was conducted to measure miR-490-3p expression in 57 pairs of OS tissues and adjacent normal tissues. (G) Pearson’s correlation coefficient showed the relationship between miR-490-3p and CCDC144NL-AS1 in the 57 OS tissues. (H) The wild-type and mutant miR-490-3p binding sequences within CCDC144NL-AS are shown. (I) HOS and Saos-2 cells were transfected with miR-490-3p mimic or miR-NC, and RT-qPCR was performed to assess the transfection efficiency. (J) Luciferase reporter assay demonstrated the binding of miR-490-3p to CCDC144NL-AS1 in OS cells. (K) RNA immunoprecipitation (RIP) assay was performed to determine the interaction between miR-490-3p and CCDC144NL-AS1 in OS cells. **P < 0.01.