TY - JOUR
T1 - LncRNA CDKN2B-AS1/miR-141/cyclin D network regulates tumor progression and metastasis of renal cell carcinoma
AU - Dasgupta, Pritha
AU - Kulkarni, Priyanka
AU - Majid, Shahana
AU - Hashimoto, Yutaka
AU - Shiina, Marisa
AU - Shahryari, Varahram
AU - Bhat, Nadeem S.
AU - Tabatabai, Laura
AU - Yamamura, Soichiro
AU - Saini, Sharanjot
AU - Tanaka, Yuichiro
AU - Dahiya, Rajvir
N1 - Funding Information:
compared to controls were also observed. Further, we studied the therapeutic potential of miR-141 in a mouse xenograft model. A significant decrease in tumor growth was observed by intratumoral delivery of miR-141 mimic compared to control over the course of experiment. Average tumor volume in the control group was 1548.4 mm3 compared to 504.7 mm3 in mice that received miR-141 mimic (Fig. 4e). In addition, miR-141 over-expression significantly induced apoptosis with a concomitant decrease in the viable population in both RCC cell lines compared to control (Fig. 5a). This pro-apoptotic role was supported by the induction of cleaved caspase-3, cleaved poly-ADP-ribose polymerase (PARP), an increase in BAX, and a decrease in BCl2 at protein levels (Fig. 5b). A significant decrease in migration (Fig. 5c) and invasion (Fig. 5d) was also observed in both RCC cell lines with miR-141 overexpression. We also examined EMT markers as change in migration and invasion are directly associated with EMT. Our results showed an increase in epithelial markers α-E-catenin and claudin with concomitant decrease in mesenchymal markers fibronectin and vimentin at both protein (Fig. 5e) and mRNA (Fig. S5) levels. Taken together, overexpression of miR-141 phenocopies the functional effects of CDKN2BAS1 inhibition in vitro and tumor growth suppression effects in vivo.
Publisher Copyright:
© 2020, The Author(s).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/8/1
Y1 - 2020/8/1
N2 - 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 pro-cancerous 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.
AB - 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 pro-cancerous 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.
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U2 - 10.1038/s41419-020-02877-0
DO - 10.1038/s41419-020-02877-0
M3 - Article
C2 - 32814766
AN - SCOPUS:85089582543
VL - 11
JO - Cell Death and Disease
JF - Cell Death and Disease
SN - 2041-4889
IS - 8
M1 - 660
ER -