Abstract

1. OATP1B1 is an important drug transporter with a complex regulatory mechanism. In this study, we wanted to investigate how LncRNA HOTAIR regulates the expression of OATP1B1 through its action on miR-206/miR-613 in HepG2 cells.

2. The expression level of LncRNA HOTAIR, miR-206/miR-613, and OATP1B1 mRNA was detected by RT-qPCR, and the OATP1B1 protein level was detected by Western blot. The competitive endogenous RNA mechanism was validated by bioinformatics analysis and a dual-luciferase reporter gene assay.

3. Our results showed that over- or under-expression of LncRNA HOTAIR correspondingly significantly increased or decreased the protein level of OATP1B1 in HepG2 cells, while no significant change in OATP1B1 mRNA level was observed. In addition, the stimulatory or inhibitory effect of LncRNA HOTAIR on OATP1B1 protein expression was correspondingly reversed by miR-206/miR-613 mimic or miR-206/miR-613 inhibitor. Finally, the reporter gene assay revealed that LncRNA HOTAIR can sponge miR-206/miR-613, which breaks the binding site of miR-206/miR-613 and OATP1B1 mRNA 3’-UTR, eliminating the stimulatory effect of LncRNA HOTAIR on OATP1B1 protein.

4. Thus, we conclude that LncRNA HOTAIR can affect the expression of OATP1B1 in HepG2 cells by sponging miR-206/miR-613, which, in turn, prevents the binding of miR-206/miR-613 and OATP1B1 mRNA 3’-UTR.

摘要

1. OATP1B1是重要的药物转运蛋白，具有复杂的调节机制。在这项研究中，我们想研究LncRNA HOTAIR如何通过其对HepG2细胞中miR-206 / miR-613的作用来调节OATP1B1的表达。

2. RT-qPCR检测LncRNA HOTAIR，miR-206 / miR-613和OATP1B1 mRNA的表达水平，Western blot检测OATP1B1蛋白水平。竞争性内源性RNA机制已通过生物信息学分析和双萤光素酶报告基因分析验证。

3. 我们的结果表明，LncRNA HOTAIR的过表达或过表达会相应地显着增加或降低HepG2细胞中OATP1B1的蛋白水平，而未观察到OATP1B1 mRNA的显着变化。另外，miR-206 / miR-613模仿物或miR-206 / miR-613抑制剂相应地逆转了LncRNA HOTAIR对OATP1B1蛋白表达的刺激或抑制作用。最后，报告基因分析表明，LncRNA HOTAIR可以使miR-206 / miR-613海绵化，从而破坏miR-206 / miR-613与OATP1B1 mRNA 3'-UTR的结合位点，消除了LncRNA HOTAIR对OATP1B1的刺激作用。蛋白。

4. 因此，我们得出结论，LncRNA HOTAIR可以通过使miR-206 / miR-613海绵化而影响HepG2细胞中OATP1B1的表达，从而阻止miR-206 / miR-613和OATP1B1 mRNA 3'-UTR的结合。