Ginsenoside Rh2 Suppresses Colon Cancer Growth By Targeting MiR-150-3p/SRCIN1/Wnt Axis

Background:Colon cancer(CC)remains one of the most frequent carcinomas of digestive system and the third most common cancer resulting in death worldwide.Micro RNA(miRNA)is small endogenous non-coding RNA,which can regulate the expression of target genes at transcription or translation level to exert complex network regulatory effects.The regulatory mechanism of miR-150-3p in colon cancer remains unclear.Ginsenoside Rh2(Rh2)extracted from ginseng has antitumor activity.Recent studies have shown that Rh2 may repress the colon cancer growth in vitro,but the underlying mechanism is still unclear.Objective:This study aims to explore the regulatory mechanism of miR-150-3p/SRCIN1/Wnt axis and the underlying mechanism of ginsenoside Rh2 in colon cancer,which may provide potential novel target for colon cancer treatment.Methods:Through the comprehensive strategy including data mining and q RT-PCR,we not only identified the relative level of miR-150-3p in colon cancer tissue and cell,but also investigated the correlation relationship between miR-150-3p and biological characteristics of colon cancer.Then the regulatory effects of miR-150-3p on proliferation and invasion of colon cancer in vitro were evaluated by CCK-8,Ed U,transwell test and wound healing assay.The expression level of the proteins involved in cell cycle and apoptosis in both colon cancer tissue and cells were assessed by Western blotting and immunohistochemistry.Furthermore,we constructed human colorectal carcinoma nude mouse model by tumor cell subcutaneous injection into right posterior flank.In animal experiment,we not only demonstrated the influence of miR-150-3p on the proliferation,migration and invasion of colon cancer in vivo,but also clarified the influence of miR-150-3p on the expression of proteins involved in cell cycle and apoptosis.Furthermore,bioinformatics,dual luciferase reporter test q RT-PCR,Western Blot and IHC were implemented to determine the mechanism of miRNA against m RNA in target gene.The relationship between Rh2 and miR-150-3p was further verified in SW620 and HCT-116 cells.Results:miR-150-3p was downregulated in colon cancer tissue and cell lines(SW480,Ca Co-2,HCT-8,SW620 and HCT-116).Functional assays indicated that the upregulation of miR-150-3p suppresses the invasion,migration and proliferation of tumor both in vivo and in vitro,promotes the expression of cell cycle-related protein(including G1/S phase checkpoint protein Cyclin D1,CDK6 and Bcl-2),decreases expression of apoptosis-related protein(including Bax and cleaved caspase 3),and vice versa.In addition,we found that SRCIN1 was upregulated in colon cancer and associated with poor prognosis.SRCIN1 was the direct target of miR-150-3p.Moreover,miR-150-3p mimic decreased Topflash/Fopflash-dependent luciferase activity,resulting in the inhibition of Wnt/β-catenin pathway.Rh2 could dose-dependently suppress the growth of colon cancer via increasing miR-150-3p expression.Rh2 alleviated the acceleration effect on Wnt/β-catenin pathway activity,cell proliferation/migration,and colony formation caused by miR-150-3p inhibition.ConclusionThe expression of miR-150-3p is down-regulated in colon cancer,and the up-regulation of miR-150-3p can inhibit the proliferation,migration and invasion of colon cancer in vivo and in vitro.The expression of SRCIN1 is up-regulated in colon cancer,and SRCIN1 is the target of miR-150-3p and negatively correlated with it.SRCIN1/miR-150-3p can inhibit the growth of colon cancer by inhibiting Wnt/ β-catenin signaling pathway.Ginsenoside Rh2 suppress colon cancer growth through the miR-150-3p /SRCIN1/Wnt axis.