Preliminary Study On The Expression And Function Of Long Non-coding RNA-AL135905.1 In Colorectal Cancer

ObjectiveTo investigate the expression of long non-coding RNA(lnc RNA)AL135905.1 in colorectal cancer(CRC)tissues and its relationship with clinicopathological features To investigate the effect of AL135905.1 on the biological function of colorectal cancer cells and to predict the potential target genes and signaling pathways that AL135905.1 may be involved in.Methods1.We screened out lnc RNA-AL135905.1 which had not been reported based on its expression through RNA-Sequencing.2.Real time fluorescence quantitative PCR(RT-q PCR)was used to validated the expression pattern of AL135905.1 in CRC tissues and cells.Chi-square test was applied to evaluate the relationship between the expression of AL135905.1 and clinicopathological features of CRC patients.3.A recombinant plasmid pcDNA3.1-AL135905.1 was constructed and transfected into CRC cell lines,Caco2 and SW480 to overexpress the expression level of AL135905.1.CCK8 assay and clone formation assay were used to evaluate the cell proliferation.Transwell chamber assay was used to investigate the cell migration and invasion.Flow cytometry was employed to detect the cell cycle and apoptosis.4.Bioinformatics analysis were applied to predict the potential target genes of AL135905.1 and explore their potential underlying mechanisms in CRC progression.Results1.The result of RT-q PCR showed that the expression of AL135905.1 was significantly down-regulated in CRC tumor tissues and cell lines.Aberrant expression of AL135905.1 was significantly relevant to TNM stage(P = 0.021),N stage(P = 0.028),perineural(P = 0.021)and venous invasion(P = 0.019)and instead,was not correlated with gender,age,differentiation,tumor size,distant metastasis,serosal invasion,KRAS mutation and BRAF-V600 E mutation(P > 0.05).2.The expression of AL135905.1 in FHC,the normal colorectal epithelial cells,was significantly higher than those CRC cell lines(P < 0.05).Overexpression of AL135905.1 inhibited cell proliferation,colony formation,migration and invasion(P < 0.05),and promoted apoptosis(P < 0.05).3.Bioinformatics analysis revealed that AL135905.1 may serve as an oncogene in CRC through the mi R-6838-5p/PHLPP2/PI3K/AKT axis.ConclusionAL135905.1 is down-regulated in CRC tumor tissues and cell lines,and AL135905.1 may play an oncogene-like role in CRC through the mi R-6838-5p/PHLPP2/PI3K/AKT axis.