The Effects Of Tenascin-C On Colorectal Cancer Stem-like Cells

Background:Tenascin-C(TNC)is an oligomeric glycoproteins with unique six-arm structure in the extracellular matrix.It has been shown to be widely expressed in numerous cancers and contribute to carcinogenesis and tumor progression.But the roles of TNC expression in colorectal cancer(CRC)cells remain unclear.Objective:To investigate the expression of TNC in CRC and its clinicopathological significance and further investigate the effects of TNC on CRC stem-like cells,and thus to be used as a molecular target for the development of treatments for CRC.Methods:We analyzed mRNA expression levels of TNC in gene expression profiling data sets from the publicly available Oncomine database.We also investigated the expression of TNC in 3 cases of colorectal human fetus tissues,100 cases of CRC.9 cases of adenoma and 35 cases of normal colorectal mucosa tissues.Thus further statistical analysis was used To demonstrate the correlation of TNC expression and clinicopathological parameters.In addition,the Kaplan-meier survival analysis was used to demonstrate the expression of TNC in evaluating the prognosis of patients with CRC.The immunofluorescent(IF)and western blotting were used to detect the expression of TNC and cancer stemness-related proteins in different CRC cell lines(HT29,HCT116)and tissues.The TNC gene was silenced by siRNA transfection and the transfection effect was verified by Western blotting.The migration and invasion assay was used to detect the effects of TNC gene silencing on migration and invasion ability of HT29 and HCT116 cells.Results:1.The validation data confirmed that mRNA expression levels of TNC were up-regulated in CRC tissues compared with their corresponding normal tissues(P<0.001).2.TNC was found to be slightly expressed in the colon of the human fetus,and the expression levels of TNC in CRC tissues was higher than adenoma and normal colorectal mucosa tissues.TNC in cancer cells was associated with tumor recurrence(P=0.034).3.The overall survival(OS)and disease free survival(DFS)rates in the TNC positive group were significantly lower than those of TNC negative group(OS:P=0.007;DFS:P=0.004).4.TNC not only positively correlated with LSD1 protein expression(P=0.007).but also was co-expressed in serial sections of CRC tissues and CRC cells(HT29.HCT116).5.The TNC knockdown group exhibited the dramatically decreased of LSD 1 expression compared to that in the blank control and control groups in the HT29 and HCT116 cell lines.Furthermore?cell invasion and migration were significantly decreased in the TNC siRNA group compared with the control groups(P<0.001)in both HT29 and HCT116 cells.Conclusion:1.High expression of TNC may be a effective target for the prognostic evaluation of CRC patients.2.TNC expressed in the CRC cell may constitute a potential CRC marker.3.TNC may plav an important role in the invasion and migration of CRC.