Na Luo1,2, Shiqin Liu1,2, Xin Li1,2, Yu Hu1,2, Kejing Zhang1,2. 1. Department of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China. 2. Clinical Research Center for Breast Cancer Control and Prevention in Hunan Province, Changsha, Hunan, China.
Abstract
Background: This study investigated the potential molecular mechanism of circular RNA HIPK3 (circHIPK3) in breast cancer (BCa). Methods: BCa cells were transfected with miR-326 mimic, miR-326 inhibitor, circHIPK3, sicircHIPK3. The expressions of circHIPK3 and miR-326 in BCa tissues and BCa cell lines were determined by RT-qPCR. Cell viability, colony formation, migration, invasion, and apoptosis of the cells were detected by CCK-8 and colony formation, wound-healing, transwell and flow cytometric assays, respectively. The relationship between circHIPK3 and miR-326 was analyzed and confirmed by circInteractome, dual-luciferase reporter, RT-qPCR, Pearson's correlation assays. Western blot and RT-qPCR were performed to determine the expressions of apoptosis-related molecules (Bcl-2, Bax, and cleaved Caspase-3) and EMT-related molecules (E-cadherin, N-cadherin, and Vimentin) in the BCa cells and tumor tissues. The tumor growth in mice was examined in a xenograft tumor model in which Ki-67 expression was determined by immunohistochemistry (IHC). Results: In BCa, the expression of circHIPK3 was up-regulated and that of miR-326 was down-regulated. CircHIPK3 knockdown inhibited the cell proliferation, invasion, and migration. MiR-326 was the direct target of circHIPK3, and was inversely correlated with circHIPK3 expression. CircHIPK3 overexpression promoted proliferation, migration, invasion, apoptosis resistance, and tumor growth and up-regulated Ki-67 expression, at the same time, the expressions of Bcl-2, N-cadherin, Vimentin were up-regulated, and those of Bax, cleaved Caspase-3 and E-cadherin were inhibited. These above expressions were partially reversed by miR-326 overexpression. Conclusion: CircHIPK3 sponges miR-326 to promote BCa growth and metastasis. The current findings provide a novel therapeutic target for treating BCa.
Background: This study investigated the potential molecular mechanism of circular RNA HIPK3 (circHIPK3) in breast cancer (BCa). Methods: BCa cells were transfected with miR-326 mimic, miR-326 inhibitor, circHIPK3, sicircHIPK3. The expressions of circHIPK3 and miR-326 in BCa tissues and BCa cell lines were determined by RT-qPCR. Cell viability, colony formation, migration, invasion, and apoptosis of the cells were detected by CCK-8 and colony formation, wound-healing, transwell and flow cytometric assays, respectively. The relationship between circHIPK3 and miR-326 was analyzed and confirmed by circInteractome, dual-luciferase reporter, RT-qPCR, Pearson's correlation assays. Western blot and RT-qPCR were performed to determine the expressions of apoptosis-related molecules (Bcl-2, Bax, and cleaved Caspase-3) and EMT-related molecules (E-cadherin, N-cadherin, and Vimentin) in the BCa cells and tumor tissues. The tumor growth in mice was examined in a xenograft tumor model in which Ki-67 expression was determined by immunohistochemistry (IHC). Results: In BCa, the expression of circHIPK3 was up-regulated and that of miR-326 was down-regulated. CircHIPK3 knockdown inhibited the cell proliferation, invasion, and migration. MiR-326 was the direct target of circHIPK3, and was inversely correlated with circHIPK3 expression. CircHIPK3 overexpression promoted proliferation, migration, invasion, apoptosis resistance, and tumor growth and up-regulated Ki-67 expression, at the same time, the expressions of Bcl-2, N-cadherin, Vimentin were up-regulated, and those of Bax, cleaved Caspase-3 and E-cadherin were inhibited. These above expressions were partially reversed by miR-326 overexpression. Conclusion: CircHIPK3 sponges miR-326 to promote BCa growth and metastasis. The current findings provide a novel therapeutic target for treating BCa.
Entities:
Keywords:
Circular RNA circHIPK3; breast cancer; growth; metastasis; miR-326
Authors: Derong Xu; Xuexiao Ma; Chong Sun; Jialuo Han; Chuanli Zhou; Matthew T V Chan; William K K Wu Journal: Cell Prolif Date: 2021-10-08 Impact factor: 6.831
Authors: Fatima Domenica Elisa De Palma; Francesco Salvatore; Jonathan G Pol; Guido Kroemer; Maria Chiara Maiuri Journal: Biomedicines Date: 2022-03-21