长链非编码RNA(lncRNA)可以作为竞争性内源RNA(ceRNA)被切断,以通过海绵微小RNA(miRNA)调节靶基因或mRNA。本研讨经由过程ceRNA机制摸索LINC01554对肝癌细胞的感化。
■选择5个显著下调的lncRNAs进行进一步验证,然后通过生物信息学,鉴定了lncRNAs的相互作用的miRNA和mRNA。通过双荧光素酶报告基因检测检测LINC01554、miR-148b-3p和EIF4E3之间的关系。之后,用pCDH-LINC01554、miR-148b-3p抑制剂和miR-148b-3p模拟物转染HCCLM3细胞。细胞活力,凋亡,迁移和侵袭通过细胞计数试剂盒-8,流式细胞仪,和Transwell分析。采用实时定量PCR(RT-qPCR)和Westernblot检测相关基因和蛋白的表达。
■LINC01554在肝癌细胞系中显著下调,并在HCCLM3细胞的胞浆中表达。LINC01554过表达抑制增殖,迁移,和HCCLM3细胞的侵袭,促进其凋亡(P<0.05)。此外,LINC01554过表达也显著增加了BAX的水平,BCL2/BAX,P53,cleaved-Caspase3,TIMP3,E-cadherin和EIF4E3(P<0.05)。通过生物信息学和双荧光素酶报告基因检测,LINC01554、miR-148b-3p和EIF4E3被证明互相感化。此外,miR-148b-3p敲低对HCCLM3细胞的影响与LINC01554过表达相似,miR-148b-3p模拟物可以逆转细胞活力的变化,凋亡,迁移,和LINC01554过表达诱导的侵袭。
■LINC01554过表达可以通过miR-148b-3p/EIF4E3抑制HCCLM3细胞的生长和转移。
UNASSIGNED: Long non-coding RNAs (lncRNAs) can be severed as competing endogenous RNAs (ceRNAs) to regulate target genes or mRNAs via sponging microRNAs (miRNAs). This study explored the effect of LINC01554 on liver cancer cells through the ceRNA mechanism.
UNASSIGNED: Five significantly down-regulated lncRNAs were selected for further verification, and then through bioinformatics, interactive miRNAs and mRNAs of lncRNAs were identified. The relationship between LINC01554, miR-148b-3p and EIF4E3 was detected by the dual luciferase reporter gene assay. Afterwards, HCCLM3 cells were transfected with pCDH-LINC01554, miR-148b-3p inhibitor and miR-148b-3p mimics. Cell viability, apoptosis, migration and invasion were measured by Cell Counting Kit-8, flow cytometer, and Transwell assays. Real-time quantitative PCR (RT-qPCR) and Western blot were used to measure the expressions of related genes and proteins.
UNASSIGNED: LINC01554 was significantly down-regulated in the liver cancer cell lines, and was expressed in the cytoplasm of HCCLM3 cells. LINC01554 overexpression inhibited proliferation, migration, and invasion of HCCLM3 cells, and promote their apoptosis (P < 0.05). Besides, LINC01554 overexpression also significantly increased the levels of BAX, BCL2/BAX, P53, cleaved-Caspase3, TIMP3, E-cadherin and EIF4E3 (P < 0.05). Through bioinformatics and dual-luciferase reporter gene assay, LINC01554, miR-148b-3p and EIF4E3 were proved to interact with each other. Furthermore, the effects of miR-148b-3p knockdown on HCCLM3 cells were similar with those of LINC01554 overexpression, and miR-148b-3p mimics could reverse the changes of cell viability, apoptosis, migration, and invasion induced by LINC01554 overexpression.
UNASSIGNED: LINC01554 overexpression could suppress the growth and metastasis of HCCLM3 cells via miR-148b-3p/EIF4E3.