MiR-3065-3p Promotes Stemness And Metastasis By Targeting CRLF1 In Colorectal Cancer

Colorectal cancer is one of the most common malignancies in the world,and the morbidity and mortality increased year by year.It has been reported that cancer stem cells(CSCs)serve as the primary drivers of tumorigenesis and tumor progression.There is an urgent need to explore novel molecules that regulate CSCs or their signatures.Increasing evidence has shown that miRNAs are involved in tumorigenesis and progression.Here,we aim to explore the regulatory effect and mechanism of miR-3065-3p on the stemness of colorectal cancer cells.The expression of miR-3065-3p in colorectal cancer and the association of miR-3065-3p expression with prognosis were analyzed using TCGA dataset or clinical cases,and we found that miR-3065-3p was overexpressed in colorectal cancer and its high expression was associated with poor prognosis of patients with colorectal cancer.Colorectal cancer cell lines were constructed to investigate the effects of miR-3065-3p on stemness and metastasis in vitro and in vivo,including the wound healing ability,Transwell migration ability,the expression of stemness transcription factors(NANOG,OCT4,SOX2),ALDH activity,sphere formation ability,xenograft tumor size and number of liver metastases in nude mice,it was found that miR-3065-3p promoted the stemness and metastasis of colorectal cancer.Further,we demonstrated that miR-3065-3p targets CRLF1 through combining database screening with dual-luciferase reporting experiments.The correlation between CRLF1 and miR-3065-3p expression was analyzed by using colorectal cancer clinical tissue samples,it turned out to be that CRLF1 expression was negatively correlated with miR-3065-3p in colorectal cancer tissues.We constructed colorectal cancer cell lines to study the effect of CRLF1 on colorectal cancer cells and found that CRLF1 inhibited the stemness and metastasis of colorectal cancer cells in vitro and in vivo.Finally,we found that CRLF1 mediated the effects of miR-3065-3p on promoting stemness of colorectal cancer cells via functional recovery experiments,including wound healing assay,Transwell migration assay,Aldefluor assay and the expression of stemness transcription factors.Based on the results of the research,we found that miR-3065-3p promoted the stemness and metastasis of colorectal cancer by targeting CRLF1.miR-3065-3p might serve as a promising prognostic marker as well as a therapeutic target for colorectal cancer.