UNASSIGNED: CircKIF5B location was investigated by RNA fluorescence in situ hybridization (RNA-FISH). For circRNA determination, RNase R treatment and Real-Time Quantitative RT-PCR (qRT-PCR) were performed. Transwell chamber assays examined the chemotactic migration and invasion of liver cancer cells.
UNASSIGNED: This study identified the circRNA circKIF5B originating from exons 1, 2, and 3 of the KIF5B gene. Importantly, we found that circKIF5B circRNA, rather than KIF5B linear mRNA, was notably upregulated in liver cancer cell lines and tissues. Moreover, we found that silencing circKIF5B markedly reduced the proliferation, invasion, and metastasis of liver cancer cells by sponging the miR-192 family, thus decreasing the expression of X-linked inhibitor of apoptosis (XIAP).
UNASSIGNED: Our data demonstrate that circKIF5B can regulate XIAP expression by sponging miR-192 and miR-215 competing for the ceRNA mechanism, indicating that circKIF5B may act as an essential upstream regulator and providing mechanistic evidence to support the view that circKIF5B/miR-192s/XIAP is a promising therapeutic target for treating liver cancer.
■通过RNA荧光原位杂交(RNA-FISH)研究CircKIF5B位置。对于circRNA测定,进行RNaseR处理和实时定量RT-PCR(qRT-PCR)。Transwell小室测定法检查了肝癌细胞的趋化迁移和侵袭。
■这项研究鉴定了源自KIF5B基因外显子1、2和3的circRNAcirkKIF5B。重要的是,我们发现circKIF5BcircRNA,而不是KIF5B线性mRNA,在肝癌细胞系和组织中显著上调。此外,我们发现沉默circKIF5B显著减少了增殖,入侵,和转移的肝癌细胞通过海绵miR-192家族,从而降低X连锁凋亡抑制剂(XIAP)的表达。
■我们的数据表明,circKIF5B可以通过攻击miR-192和miR-215竞争ceRNA机制来调节XIAP表达,这表明circKIF5B可能充当重要的上游调节因子,并提供了机械证据来支持circKIF5B/miR-192s/XIAP是治疗肝癌的有希望的治疗靶点的观点.