LncRNA CDKN2B-AS1/miR-141/cyclin D network regulates tumor progression and metastasis of renal cell carcinoma

Abstract

The molecular heterogeneity of renal cell carcinoma (RCC) complicates the therapeutic interventions for advanced metastatic disease and thus its management remains a significant challenge. This study investigates the role of the lncRNA CDKN2B-AS1 and miR-141-3p interactions in the progression and metastasis of kidney cancer. Human renal cancer cell lines (ACHN and Caki1), normal RPTEC cells, tissue cohorts, and a series of in vitro assays and in vivo mouse model were used for this study. An overexpression of CDKN2B-AS1 was observed in RCC compared to normal samples in TCGA and our in-house SFVAMC tissue cohorts. Reciprocally, we observed reduced expression of miR-141 in RCC compared to normal in the same cohorts. CDKN2B-AS1 shares regulatory miR-141 binding sites with CCND1 and CCND2 genes. Direct interactions of CDKN2B-AS1/miR-141/Cyclin D1-D2 were confirmed by RNA immunoprecipitation and luciferase reporter assays indicating that CDKN2B-AS1/miR-141/Cyclin D1-D2 acts as a ceRNA network in RCC. Functionally, attenuation of CDKN2B-AS1 and/or overexpression of miR-141 inhibited proliferation, clonogenicity, migration/invasion, induced apoptosis in vitro and suppressed tumor growth in xenograft mouse model. Further, overexpression of CDKN2B-AS1 is positively correlated with poor overall survival of RCC patients. Expression of miR-141 also robustly discriminated malignant from non-malignant tissues and its inhibition in normal RPTEC cells induced procancerous characteristics. CDKN2B-AS1 attenuation or miR-141 overexpression decreased CCND1/CCND2 expression, resulting in reduced RAC1/pPXN that are involved in migration, invasion and epithelial-mesenchymal transition. This study, for the first time, deciphered the role of CDKN2B-AS1/miR-141/Cyclin D axis in RCC and highlights this network as a promising therapeutic target for the regulation of EMT driven metastasis in RCC.

LncRNA CDKN2B-AS1 stabilized by IGF2BP3 drives the malignancy of renal clear cell carcinoma through epigenetically activating NUF2 transcription

“…In addition, we further verified the effects of CDKN2B-AS1 on KIRC carcinogenesis in vivo and confirmed its prognostic capability by multivariate analysis. Hence, our findings, together with those of other studies 22 , 23 , indicate that CDKN2B-AS1 drives the malignancy of KIRC in vitro and in vivo, and has the potential to serve as an independent prognostic factor for worse KIRC progression.…”

“…However, lncRNAs located in the nucleus can participate in the gene regulation at the chromatin modification and gene transcription levels 36 . Although the latest study 23 showed that CDKN2B-AS1 can function as a ceRNA to regulate cyclin-D1/D2 expression, resulting in RCC aggressiveness, by using cell cytoplasm/nuclear fractionation assays, we found that CDKN2B-AS1 is mainly localized in the nucleus, as in previous reports on other tumors 30 , 37 , 38 , and indicating that CDKN2B-AS1 might regulate NUF2 expression via a transcriptional mechanism. Since CDKN2B-AS1 was reported to cooperate with chromatin-modifying enzymes CBX7/PRC1 or SUZ12/PRC2, it affects histone H3K27me3 enrichment in the promoter region, thus activating epigenetic activation or gene silencing 37 , 39 .…”

“…Accumulating evidence has demonstrated that CDKN2B-AS1 is upregulated in a variety of human cancers, such as bladder, gastric, and breast cancers, and serves as an oncogene that plays important roles in tumor progression 19 . Although two other studies have investigated the role of CDKN2B-AS1 in RCC and found as well that it is overexpressed 22 , 23 , they were limited to verify the effects of CDKN2B-AS1 in vitro experiments, and did not discuss the reasons for the upregulation of it. In the present study, we demonstrated that CDKN2B-AS1 was elevated and correlated with poor clinical outcomes in KIRC, and promoted cell proliferation, migration, and invasion in vitro.…”

“…Moreover, CDKN2B-AS1 interacts with signaling pathways in cancers, such as p38 MAPK, PI3K/AKT, mTOR, ATM-E2F1, and TGF-β 19 . Therefore, it is not surprising that CDKN2B-AS1 can promote RCC carcinogenesis via the β-catenin pathway or function as a ceRNA for miR-141 to regulate cyclin-D1/D2 expression 22 , 23 . We used multiple open-source large datasets to identify the potential target genes regulated by CDKN2B-AS1, and also proved in vivo and in vitro that CDKN2B-AS1 affects KIRC tumor growth and metastasis, at least partly through NUF2, which can also serve as an oncogene to promot tumor progression 32 , 33 .…”

lncRNA FDNCR promotes apoptosis of granulosa cells by targeting the miR-543-3p/DCN/TGF-β signaling pathway in Hu sheep

“… 6 Recently, numerous lncRNAs have been identified in humans and other organisms that play diverse roles in regulating various phenomenon, such as cell proliferation and apoptosis, 7 , 8 cell development, 9 cell differentiation, 10 and development of certain diseases. 11 , 12 Specifically, lncRNAs are involved in regulating spermatogenesis, 13 , 14 steroidogenesis, 15 embryo implantation 16 and development, 17 follicular development, 18 oocyte maturation, 19 GC apoptosis and proliferation, 20 , 21 and reproductive diseases, 22 , 23 suggesting their importance in reproduction. 24 However, only a few lncRNAs have been identified in domestic animals.…”