miR-548b-3p regulates the expression of decoy receptor 3 and influences the malignant biological traits of hepatoma cells / 国际外科学杂志

ABSTRACT

Objective:To investigate whether miR-548b-3p can regulate the malignant biological traits of hepatocellular carcinoma cells by down-regulating the expression of decoy receptor 3( DcR3). Methods:The miR-548b-3p levels in 5 common hepatoma cell lines Hep3b, HepG2, Huh7.5.1, PLC and 97H were detected by real-time fluorescence quantitative PCR. The experimental hepatoma cell lines were purchased from Shanghai Cancer Institute. The subcutaneous tumor formation experiment in nude mice included two parts: miR-548b-3p over expression group and down expression group. miR-548b-3p over expression experiment included Huh7.5.1- miR-548b-3p cell group and Huh7.5.1-NC cell group. The experimental group with low expression of miR-548b-3p included PLC- miR-548b-3p-sponge group and PLC-NC cell group, with 5 nude mice in each group. The nude mice used in the experiment were male Balb/c strain, body weight 20-25 g, 4-6 weeks old. Starbase software was used to predict whether there were binding sites between miR-548b-3p and DcR3. Dual luciferase reporting assay to verify whether DcR3 is the target gene of miR-548b-3p. Rescue experiments were conducted to verify whether the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Statistical analysis was performed using Graphpad Prism 9. Measurement data with normal distribution were expressed as mean±standard deviation( ± s), and t-test was adopted for comparison between general measurement data groups. Results:The relative expression level of 2 -ΔΔCt was used as the parameter for real-time fluorescence quantitative PCR comparison. Hep3b expression level was 1, HepG2 expression level was 0.902, Huh7.5.1 expression level was 0.712, PLC expression level was 1.293, and 97H expression level was 0.818. The final results showed that, The highest expression of miR-548b-3p was in PLC cells, and the lowest expression was in Huh7.5.1 cells. The subcutaneous tumor formation experiment of nude mice showed that after 4 weeks, the tumor volume of Huh7.5.1- miR-548b-3p was(444.77±142.34) mm 3, and that of Huh7.5.1-NC was(918.80±139.21) mm 3. The tumor volume of PLC- miR-548b-3p-sponge was(407.49±58.50) mm 3, and that of PLC-NC was(218.62±47.55) mm 3. The binding sites of miR-548b-3p and DcR3 were predicted by Starbase software. Dual luciferase assay showed that DcR3 was the target gene of miR-548b-3p. Rescue experiments verified that the effects of miR-548b-3p on the proliferation, invasion and migration of liver cancer cells were realized through DcR3. Conclusion:miR-548b-3p can regulate the proliferation, invasion, migration and other malignant biological traits of liver cancer cells, and it is achieved by down-regulating the expression level of DcR3.