Methods: c9orf140, EZH2, beta-catenin, STAT5 and pSTAT5 expressions were measured by Western Blot and immunohistochemical staining. To establish stable cells, CRC cells were infected with c9orf140 shRNA, c9orf140 overexpression or control lentivirus. Stable cell lines with different protein or shRNA expressions were introduced into nude mice respectively via tail veins injection. The photos were taken by the NightOWL II LB 983 in vivo Imaging system. Chromatin Immunoprecipitation (ChIP) and Luciferase reporter gene assay were carried
out according to the protocol of manufactures. Results: there is almost no expression of C9orf140 in a normal human colon epithelial find more cell line, CRL-1790, but the expression of C9orf140 is significantly increased in all CRC cell line especially in those highly invasive CRC cells. Immunofluorence analysis showed that C9orf140 is mainly detected in the cytoplasm of CRC HCT116 and HT29 cells. Immunohistochemical analysis of 150 patients’ tissue sections showed that the staining of C9orf140 was found
at higher expression levels in CRC samples than normal samples. Furthermore, the expression of C9orf140 was gradually increased from well differentiated to poor differentiated in CRC tissues. Knockdown of C9orf140 dramatically increased E-cadherin expression and decreased N-cadherin expression. Knockdown of C9orf140 significantly reduced the CRC cell invasion. More lung metastasis and shorter overall survival were detected after overexpression of C9orf140 compared with control in vivo. Knockdown of C9orf140 dramatically increased overall survival and decreased metastasis of lung in vivo. Western Blot RAD001 molecular weight data showed that C9orf140 expressions may be regulated by EZH2, STAT5 and β-catenin. Moreover, Immunoprecipitation, ChIP and Luciferase reporter gene assay 3-oxoacyl-(acyl-carrier-protein) reductase showed that EZH2, STAT5 and β-catenin may colocalized together and directly bind to the promoter of C9orf140 gene to upregulate this gene expression in CRC cells. Conclusion: we firstly revealed the role of C9orf140 in the metastatic progression of CRC in vitro and in vivo.
C9orf140-induced CRC cell migration and invasion may depend on promote the progression of EMT. Furthermore, we firstly identified that c9orf140 expression may be directly regulated by EZH2, STAT5 and β-catenin. Our data may provide potential targets to prevent and/or treat aggressive CRC. Key Word(s): 1. C9orf140; 2. CRC invasion; 3. STAT5; Presenting Author: SARAVANAN. ARJUNAN Additional Authors: NAGESHWAR REDDY, MANU TANDAN, RUPA BANERJEE Corresponding Author: SARAVANAN. ARJUNAN Affiliations: None Objective: Inadequate bowel preparation leads to incomplete examination, cancellation, missed lesion and increased complication./Objective of the study to assess the relationship between frequency of bowel opening and quality of bowel preparation. Methods: Single center prospective observational study.