Dysregulated And Effects Of C14orf28 In Colorectal Cancer Cells

Objective:Colorectal cancer(CRC)is one of the common malignant tumor.Many reasons promote the process of colorectal cancer,including the abnormal of micrRNAs(miRNA).The protein level of C14orf28 is increased in the mental health disorders,and our lab discovered that C14orf28 can be regulated by miRNAs.The main purpose of this paper is to explore the C14orf28 function in colorectal cancer and the miRNAs can regulate the C14orf28.Thus,our research found the function of C14orf28 which may contribute to the molecular mechanism of colorectal cancer.Methods:RT-qPCR was used to examine C14orf28 expression level in 20 pairs of colorectal cancer tissues.MTT,colony formation assays and migration,invasion assays uncovered the C14orf28 effect on cell activity,proliferation,migration and invasion.Using apoptosis assays to detect C14orf28 influence on colorectal cancer cell apoptosis ability.Bioinformatics analyzed the miRNAs bind to the C14orf28 3 'UTR and combination of miRNAs known functions,we selected miR-519 d for further researches.To confirm C14orf28 can actually binded by miR-519 d,we constructed a C14orf28 3 'UTR EGFP report vector containing miR-519 d binding sites and a 3' UTR mut EGFP report vector containing the mutation of miR-519 d binding sites.EGFP report vector assays uncovered that C14orf28 can binded by miR-519 d directly;RT-qPCR and western blot were used to determine if miR-519 d directly binds C14orf28 in mRNA and protein levels.We also examined miR-519 d expression level in 20 pairs of colorectal cancer tissues using RT-qPCR,and correlation analysis was used to test the correlation of C14orf28 and miR-519 d.To further study the function of miR-519 d,MTT,colony formation assays,apoptosis assays,migration and invasion assays applied to explore the effect of miR-519 d in colorectal cancer cells.Finally,rescue experiments demonstrated that C14orf28 was targeted by miR-519 d to play roles.Results:Compared with the adjacent tissues,C14orf28 expression level was higher in colorectal cancer tissues.In THC-8307 and HCT116 cells,C14orf28 can promote the activity of cell growth,proliferation but inhibited apoptosis of colorectal cancer cells.Overexpression of C14orf28 can promote migration and invasion ability of CRC cells,and knock down C14orf28 get the opposite results.Using EGFP report vector experiments proved that miR-519 d can directly target C14orf28.RT-qPCR and western blot were used to determine miR-519 d directly targeted C14orf28 and downregulated the mRNA and protein level of C14orf28.Subsequently,the expression level of miR-519 d was downregulated in colorectal cancer tissues compared with the adjacent tissues and the correlation analysis between C14orf28 and miR-519 d showed that miR-519 d negatively regulated C14orf28.In THC-8307 and HCT116 cells,overexpression of miR-519 d plasmid can inhibit cell viability and proliferation ability and promote apoptosis of CRC cells.Overexpression of miR-519 d plasmid can inhibit colorectal cancer cells migration,invasion ability.Rescue experiments confirmed that C14orf28 was targeted by miR-519 d to restrain the malignant activity in colorectal cancer cells.Conclusion:our results indicated that C14orf28 was targeted by miR-519 d,miR-519d/C14orf28 axis can inhibit CRC cells oncogenic but promotes apoptosis,which may provide new insights into the molecular mechanism and potential bilmarkers for colorectal cancer.