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Abstract
MicroRNAs are noncoding RNA molecules that posttranscriptionally regulate gene expression. The aim of this study was to determine the role of microRNA-21 in cholangiocarcinomas and its relationship to cholangiocarcinoma RBE cell capacity for invasion and metastasis. MicroRNA-21 expression was investigated in 41 cases of cholangiocarcinoma samples by in situ hybridization and real-time PCR. Influence on cholangiocarcinoma cell line invasion and metastasis was analyzed with microRNA-21 transfected cells. In addition, regulation of reversion-inducing-cysteine-rich protein with kazal motifs (RECK) by microRNA-21 was elucidated to identify mechanisms. In situ hybridization and real-time quantitative PCR results for patients with lymph node metastasis or perineural invasion showed significantly high expression of microRNA-21 (P<0.05). There was a dramatic decrease in cholangiocarcinoma cell line invasion and metastasis ability after microRNA-21 knockdown (P<0.05). However, overexpression significantly increased invasion and metastasis (P<0.05). Real-time PCR and Western-blot analysis showed that microRNA-21 could potentially inhibit RECK expression in RBE cells. Survival analysis showed that patients with higher expression levels of microRNA-21 more often had a poor prognosis (P<0.05). MicroRNA-21 may play an important role in cholangiocarcinoma invasion and metastasis, suggesting that MicroRNA-21 should be further evaluated as a biomarker for predicting cholangiocarcinoma prognosis.
Highlights
Cholangiocarcinoma (CCA) is a highly malignant tumor derived from bile duct epithelial cells (Sawanyawisuth, 2009)
RBE cells were suitable for the invasion and metastasis assay, because they showed good invasion to the Matrigel membranes.The cell invasion and metastasis assay showed that knockdown of microRNA-21 resulted in decreased RBE cell invasion and metastasis rate compared with the control cells (P
We found that RBE cells transfected with microRNA-21-IN had dramatically increased RECK levels by 10 fold, compared with the control cells (P
Summary
Cholangiocarcinoma (CCA) is a highly malignant tumor derived from bile duct epithelial cells (Sawanyawisuth, 2009). The role for MicroRNAs in the establishment and progression of cholangiocarcinoma has become evident,and some miRNAs have been identified as oncogenes or tumor suppressor genes in cholangiocarcinoma (Li et al, 2012; Zhang et al, 2012). RECK has been known to contain the microRNA-21 binding site, which acts as a tumor suppressor gene regulating various aspects associated with cancer cell invasion, metastasis (Liu et al, 2010; Han et al, 2011). Methods: MicroRNA-21 expression was investigated in 41 cases of cholangiocarcinoma samples by in situ hybridization and real-time PCR. Results: In situ hybridization and real-time quantitative PCR results for patients with lymph node metastasis or perineural invasion showed significantly high expression of microRNA-21 (P
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