The Mechanism Research Of Quercetin Inhibiting The Progression Of Hepatocellular Carcinoma By Targeting Human Antigen R

Objective Investigating quercetin in reducing the stability of non-coding RNA LINC01123 and inhibiting the proliferation and metastasis of hepatocellular carcinoma cells(HCC)by targeting human antigen R(HuR),further studying the anti-tumor mechanism of quercetin and provide a new idea for the treatment of HCC with traditional Chinese medicine.Methodsl.We proved that quercetin could inhibit the proliferation of HCC cells and induce the apoptosis of HCC cells by performing CCK-8 assay,EdU assay,transwell assay,plate clone formation assay and flow cytometry on HCC cells treated with different concentrations of quercetin.2.We proved that quercetin could inhibit the migration and invasion of HCC cells by performing wound healing assay and transwell assay on HCC cells treated with different concentrations of quercetin.3.We observed whether quercetin could inhibit the progression of tumor in vivo.4.We studied the relationship between prognosis and the expression of LINC01123 in hepatocellular carcinoma by bioinformatic analysis and clinical data analysis.5.We investigate the role of LINC01123 in promoting tumor growth,migration and invasion in vivo and in vitro by performing CCK-8 assay,EdU assay,transwell assay and nude mouse transplantation tumor experiment on HCC cells that overexpress or knock down LINC01123.6.We identified the target of LINC01123 by bioinformatic analysis,RNA immunoprecipitation(RIP)assay and dual-luciferase reporter assay.7.We identified the target of miR-34a-5p by bioinformatic analysis and dual-luciferase reporter assay.8.We proved that LINC01123 could promote the progression of HCC through miR-34a-5p/TUFT1 axis by TUFT1 rescue experiment.9.We explored the correlation between the expression level of HuR and LINC01123 by bioinformatic analysis and the RIP experiment was used to prove whether they could be combined directly.10.We explored whether HUR can promote the expression of LINC01123 by increasing its stability by detect the stability and expression of LINC01123 in HCC cells that overexpress or knock down HuR.11.We explored whether HUR could be the target of quercetin through bioinformatic analysis.12.We explored whether quercetin down-regulates the expression of LINC01123 by targeting HuR.Results1.Quercetin can inhibit the proliferation of Hep3B,Huh7 and MHCC97h cells in a concentration-gradient dependent manner.2.Quercetin can inhibit the metastasis of Hep3B,Huh7 and MHCC97h cells in a concentration-gradient dependent manner.3.Quercetin inhibits tumor growth in vivo.4.LINC01123 is highly expressed in HCC tissues and is positively correlated with poor prognosis of patients.5.The knock-down of LINC01123 significantly inhibited the proliferation,invasion and migration of Huh7 cells and the overexpression of LINC01123 significantly promoted the proliferation,metastasis and invasion of HepG2 and Hep3B cells.Meanwhile,the silencing of LINC01123 can inhibit tumor proliferation in vivo.6.It was found that LINC01123 was positively correlated with the expression level of TUFT1 by bioinformatic analysis.RIP and dual-luciferase reporter assay proved that LINC01123 was directly bound to miR-34-5p.7.It was found that LINC01123 was negatively correlated with the expression level of miR-34a-5p by bioinformatic analysis.Dual-luciferase reporter assay proved that miR-34-5p was directly bound to TUFT1.8.Overexpression of miR-34-5p can reverse the effects of LINC01123 on promoting the proliferation,migration and invasion of HCC.Overexpression of TUFT1 can reverse the effects of miR-34-5p on promoting the proliferation,migration and invasion of HCC.9.HuR was positively correlated with LINC01123 in HCC tissues.RIP proved that HuR could directly bound to LINC01123.10.The expression of LINC01123 can be regulated in the same direction by overexpression or knock-down of HuR expression and the knock-down of HuR can reduce the stability of LINC01123.11.It was proved that HuR was a potential target of quercetin through bioinformatic analysis and molecular docking.12.The overexpression of HuR can reverse the inhibition of proliferation and metastasis caused by different concentrations of quercetin in Hep3B,Huh7 and MHCC97h cells.Conclusion In this study,we found that LINC01123 was highly expressed in HCC tissue and positively correlated with poor prognosis of HCC patients.It can promote the proliferation and metastasis of HCC cells through miR-34a-5p/TUFT1 axis.HuR can increase the stability of LINC01123 and promote its expression by directly binding to LINC01123.However,quercetin can reduce the stability of LINC01123 and inhibit the proliferation and metastasis of HCC cells by targeting HuR.This study first discovered the mechanism of quercetin in the treatment of HCC through lncRNA and provided a new idea for the treatment of HCC with traditional Chinese medicine.