Mechanisum Study About LncRNA AK096729 Contributes To CRC Progress By Stimulating MTOR Signaling Pathway

BACKGROUND AND AIMS: The colorectal cancer is affected by both heredity and environment.Long non-coding RNAs(lnc RNAs)play a role in carcinogenesis.But the specific mechanism has not been studied yet.In this study,we identified a novel lnc RNA,AK096729,by integrated analysis in colorectal cancer tissues compared to adjacent tissues.We have studied its mechanism of how to promote the growth of colorectal cancer cells.Furthermore,we have looked for its origin to explain the phenomenon why AK096729 was highly expressed in tumor tissues.In recent years,the correlation and mechanism between host gut microbiota and colorectal cancer have become the newest research hot dots.Enterotoxigenic Bacteroides fragilis(ETBF),the pathogenic bacteria in human gut,is considered to be carcinogenic.In this study,we will discuss the correlation between lnc RNA AK096729 and ETBF.Probably,we could provide a new target for the prevention and treatment of colorectal cancer.METHODS: We tested 5 pairs of colorectal tumor tissues and adjacent tissues by lnc RNA chip.The result showed that AK096729 expression was obviously increased from paracancerous tissue to colorectal cancer tissue.We collected 96 pairs of CRC tissues to identify AK096729 expression and its association with tumor size,TNM stages,and prognosis of patients through Real-time PCR assay and SPSS19.0 software.HCT116 and DLD-1 cells were respectively transfected with AK096729 plasmids and si RNA in order to analyze the biological functions of AK096729 involved of cell proliferation,cell cycle and cell apoptosis.We studied the signal pathway of AK096729 by Western blot analysis.Furthermore,we analyzed the correlation between AK096729 and ETBF in CRC tissues.We then detected the m RNA level of AK096729 after ETBF infection in HCT116 and DLD-1 cells by real-time PCR.We also tested the signal pathway after ETBF infection in CRC cells by Western blot.RESULT: High levels of AK096729 expression were associated with tumor size and poor prognosis in colorectal cancer.Further,AK096729 affected colorectal cancer cell proliferation?cell cycle?apoptosis in multiple models.The result showed that AK096729 promoted colorectal cancer cell proliferation,cell cycle,and inhibit apoptosis.Mechanistically,AK096729 activated m TOR signal pathway in colorectal cancer cells.The target genes Cyclin D1 were high-lighted.The pathway was highly suppressed by m TOR supressor Rapamycin.We also found that the ETBF gathered more densely in colorectal tissue than adjacent normal tissue.Also,they were closely related to the expression level of AK096729 in CRC tissue.Furthermore,AK096729 expression in HCT116 and DLD-1 cells was prominently evaluated after infection by ETBF.Simultaneously,the m TOR signal pathway was also activated by ETBF infection in CRC cells.CONCLUSION: Lnc RNA AK096729 is mechanistically,functionally and clinically oncogenic in colorectal cancer.It can promote cell proliferation,shorten cell cycles and inhibit cell apoptosis.Highly expression of AK096729 in CRC tissue will indicate poorer prognosis in CRC patients.ETBF can reglulate the expression of AK096729 and activate the m TOR signal pathway to accelerate CRC occurrence and development.Targeting AK096729 and eradication of ETBF may be meaningful for treating patients with colorectal cancer.