Abstract
Extensive stromal interaction is one reason for the dismal outcome of biliary tract cancer (BTC) patients. Epithelial to mesenchymal transition (EMT) is involved in tumor invasion and metastasis and is partly regulated by microRNAs (miRs). This study explores the expression of anti-EMT miR200 family (miR141, −200a/b/c, −429) and miR205 as well as the EMT-related proteins E-cadherin and vimentin in a panel of BTC cell lines and clinical specimens by quantitative real-time polymerase chain reaction, Western blot and immunohistochemistry, respectively. MicroRNA expression was correlated to (i) the expression patterns of E-cadherin and vimentin; (ii) clinicopathological characteristics; and (iii) survival data. MicroRNA-200 family and miR205 were expressed in all BTC cells and clinical specimens. E-cadherin and vimentin showed a mutually exclusive expression pattern in both, in vitro and in vivo. Expression of miR200 family members positively correlated with E-cadherin and negatively with vimentin expression in BTC cells and specimens. High expression of miR200 family members (but not miR205) and E-cadherin was associated with longer survival, while low miR200 family and high vimentin expression was a predictor of unfavorable survival. Overall, the current study demonstrates the relevance of the miR200 family in EMT of BTC tumors and suggests these miRs as predictors for positive outcome.
Highlights
MicroRNAs are small non protein-coding RNA molecules with a length of about 21–25 nucleotides that play a key role in the regulation of gene expression
Sncii.n20v1i6t,r1o7,s2y0s53tem of a panel of biliary tract cancer (BTC) cell lines (n = 8), expression of members of the mi3Ro2f0105 family and miR205 could be detected in all cell lines, with the EGI-1, GBC, MzChA-1, SkChA-1 and TTFFKK--11 cceelllllilnineessshshoowwininggrerlealtaivtievleylyhihghigmh imRieRxperxepsrseiossnioanndanthde tBhDe CB,DCCC,SCWC-S1Wan-d1 ManzdCMhAz-C2hcAel-l2licneelsl slihnoews sinhgowcoinmgpcaormabplyarlaobwlyerloewxperreessxipornes(Fsiiognur(eFi1gAu,rBe).1A,B)
Similar results were obtained on the mRNA level for both of these markers (Figure 1D)
Summary
MicroRNAs (miRs) are small non protein-coding RNA molecules with a length of about 21–25 nucleotides that play a key role in the regulation of gene expression. Almost 30,000 miR species are known [2], and it is clear today that aberrant miR expression plays a crucial role in development and progression of various types of cancers [3,4]. Epithelial to mesenchymal transition (EMT) is a key process in the progression of cancer, in which epithelial cells lose their polarity and gain mesenchymal traits, including the ability to detach from the primary tumor, invade surrounding tissue and eventually form metastases [5,6,7]. Expression of E-cadherin was increased, leading to reduction of the invasive potential of cancer cells and making miR200 a potent tumor suppressor [6,9]
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