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A majority of early colorectal cancers ( CRC s) with submucosal invasion undergo surgical operation, despite a very low incidence of lymph node metastasis. Our study aimed to identify micro RNA s (mi RNA s) specifically responsible for lymph node metastasis in submucosal  CRC s. Micro RNA  microarray analysis revealed that miR‐100 and miR‐125b expression levels were significantly lower in  CRC  tissues with lymph node metastases than in those without metastases. These results were validated by quantitative real‐time  PCR  in a larger set of clinical samples. The transfection of a miR‐100 or miR‐125b inhibitor into colon cancer  HCT 116 cells significantly increased cell invasion, migration, and  MMP  activity. Conversely, overexpression of miR‐100 or miR‐125b mimics significantly attenuated all these activities but did not affect cell growth. To identify target m RNA s, we undertook a gene expression array analysis of miR‐100‐silenced  HCT 116 cells as well as negative control cells. The Ingenuity Pathway Analysis, TargetScan software analyses, and subsequent verification of m RNA  expression by real‐time  PCR  identified mammalian target of rapamycin (m TOR ) and insulin‐like growth factor 1 receptor ( IGF 1R) as direct, and Fas and X‐linked inhibitor‐of‐apoptosis protein ( XIAP ) as indirect candidate targets for miR‐100 involved in lymph node metastasis. Knockdown of each gene by si RNA  significantly reduced the invasiveness of  HCT 116 cells. These data clearly show that downregulation of miR‐100 and miR‐125b is closely associated with lymph node metastasis in submucosal  CRC  through enhancement of invasion, motility, and  MMP  activity. In particular, miR‐100 may promote metastasis by upregulating m TOR ,  IGF 1R, Fas, and  XIAP  as targets. Thus, miR‐100 and miR‐125b may be novel biomarkers for lymph node metastasis of early  CRC s with submucosal invasion.

Downregulation of micro RNA ‐100/micro RNA ‐125b is associated with lymph node metastasis in early colorectal cancer with submucosal invasion