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microRNA-182 targets special AT-rich sequence-binding protein 2 to promote colorectal cancer proliferation and metastasis.

Yang MH, Yu J, Jiang DM, Li WL, Wang S, Ding YQ - J Transl Med (2014)

Bottom Line: We found that miR-182 expression is increased in CRC cells that originated from metastatic foci and human primary CRC tissues with lymph node metastases.Further research showed that miR-182 could directly target the 3'untranslated region (3'UTR) of SATB2 mRNA and subsequently repress both the mRNA and protein expressions of SATB2, which we identified in previous studies as a CRC metastasis-associated protein.Investigations of possible mechanisms underlying these behaviors induced by miR-182 revealed that miR-182 induced epithelial-mesenchymal transition (EMT) by modulating the expression of key cellular molecules in EMT.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. shuangw@126.com.

ABSTRACT

Background: Increasing evidence has revealed that microRNAs (miRNA) played a pivotal role in regulating cancer cell proliferation and metastasis. The deregulation of miR-182 has been identified in colorectal cancer (CRC). However, the role and mechanism of miR-182 in CRC have not been completely understood yet.

Methods: The expression levels of miR-182 in CRC tissues and CRC cell lines were examined by performing stem-loop quantitative RT-PCR. The stable over-expression miR-182 cell lines and control cell lines were constructed by lentivirus infection. Subsequently, CCK-8 assay, plate colony formation assay, cell migration, invasion assay and experimental animal models were performed to detect the biological functions of miR-182 in vitro and in vivo. A luciferase reporter assay was conducted to confirm target associations. Western blot and immunohistochemical analysis were performed to examine the expression changes of molecular markers that are regulated by miR-182.

Results: We found that miR-182 expression is increased in CRC cells that originated from metastatic foci and human primary CRC tissues with lymph node metastases. The ectopic expression of miR-182 enhanced cell proliferation, invasion, and migration in vitro. Stable overexpression of miR-182 also facilitated tumor growth and metastasis in vivo too. Further research showed that miR-182 could directly target the 3'untranslated region (3'UTR) of SATB2 mRNA and subsequently repress both the mRNA and protein expressions of SATB2, which we identified in previous studies as a CRC metastasis-associated protein. Restoring SATB2 expression could reverse the effects of miR-182 on CRC cell proliferation and migration. Investigations of possible mechanisms underlying these behaviors induced by miR-182 revealed that miR-182 induced epithelial-mesenchymal transition (EMT) by modulating the expression of key cellular molecules in EMT.

Conclusions: Our results illustrated that the up-regulation of miR-182 played a pivotal role in CRC tumorigenesis and metastasis, which suggesting a potential implication of miR-182 in the molecular therapy for CRC.

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Over-expression of miR-182 induced epithelial mesenchymal transition in vitro and in vivo. (A) The expression of epithelial markers, mesenchymal marker, Snail, and SATB2 were compared by western blot analysis in SW480/miR-con, SW480/miR-182 and SW480/miR-182 with exogenetic overexpression of SATB2. Tublin was used as a loading control. (B) Immunohistochemistry staining of indicated proteins of the tumors in caecum terminus of nude mice.
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Figure 7: Over-expression of miR-182 induced epithelial mesenchymal transition in vitro and in vivo. (A) The expression of epithelial markers, mesenchymal marker, Snail, and SATB2 were compared by western blot analysis in SW480/miR-con, SW480/miR-182 and SW480/miR-182 with exogenetic overexpression of SATB2. Tublin was used as a loading control. (B) Immunohistochemistry staining of indicated proteins of the tumors in caecum terminus of nude mice.

Mentions: We assessed the epithelial and mesenchymal markers with western blot technique. As expected, the expression levels of Snail and mesenchymal maker Vimentin were strikingly upregulated in miR-182-overexpression cells, whereas SATB2 and epithelial marker E-cadherin levels were downregulated. Thereafter, SATB2 overexpression could significantly attenuate the expression changes of the above markers that are associated with miR-182 (FigureĀ 7A).


microRNA-182 targets special AT-rich sequence-binding protein 2 to promote colorectal cancer proliferation and metastasis.

Yang MH, Yu J, Jiang DM, Li WL, Wang S, Ding YQ - J Transl Med (2014)

Over-expression of miR-182 induced epithelial mesenchymal transition in vitro and in vivo. (A) The expression of epithelial markers, mesenchymal marker, Snail, and SATB2 were compared by western blot analysis in SW480/miR-con, SW480/miR-182 and SW480/miR-182 with exogenetic overexpression of SATB2. Tublin was used as a loading control. (B) Immunohistochemistry staining of indicated proteins of the tumors in caecum terminus of nude mice.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4020308&req=5

Figure 7: Over-expression of miR-182 induced epithelial mesenchymal transition in vitro and in vivo. (A) The expression of epithelial markers, mesenchymal marker, Snail, and SATB2 were compared by western blot analysis in SW480/miR-con, SW480/miR-182 and SW480/miR-182 with exogenetic overexpression of SATB2. Tublin was used as a loading control. (B) Immunohistochemistry staining of indicated proteins of the tumors in caecum terminus of nude mice.
Mentions: We assessed the epithelial and mesenchymal markers with western blot technique. As expected, the expression levels of Snail and mesenchymal maker Vimentin were strikingly upregulated in miR-182-overexpression cells, whereas SATB2 and epithelial marker E-cadherin levels were downregulated. Thereafter, SATB2 overexpression could significantly attenuate the expression changes of the above markers that are associated with miR-182 (FigureĀ 7A).

Bottom Line: We found that miR-182 expression is increased in CRC cells that originated from metastatic foci and human primary CRC tissues with lymph node metastases.Further research showed that miR-182 could directly target the 3'untranslated region (3'UTR) of SATB2 mRNA and subsequently repress both the mRNA and protein expressions of SATB2, which we identified in previous studies as a CRC metastasis-associated protein.Investigations of possible mechanisms underlying these behaviors induced by miR-182 revealed that miR-182 induced epithelial-mesenchymal transition (EMT) by modulating the expression of key cellular molecules in EMT.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. shuangw@126.com.

ABSTRACT

Background: Increasing evidence has revealed that microRNAs (miRNA) played a pivotal role in regulating cancer cell proliferation and metastasis. The deregulation of miR-182 has been identified in colorectal cancer (CRC). However, the role and mechanism of miR-182 in CRC have not been completely understood yet.

Methods: The expression levels of miR-182 in CRC tissues and CRC cell lines were examined by performing stem-loop quantitative RT-PCR. The stable over-expression miR-182 cell lines and control cell lines were constructed by lentivirus infection. Subsequently, CCK-8 assay, plate colony formation assay, cell migration, invasion assay and experimental animal models were performed to detect the biological functions of miR-182 in vitro and in vivo. A luciferase reporter assay was conducted to confirm target associations. Western blot and immunohistochemical analysis were performed to examine the expression changes of molecular markers that are regulated by miR-182.

Results: We found that miR-182 expression is increased in CRC cells that originated from metastatic foci and human primary CRC tissues with lymph node metastases. The ectopic expression of miR-182 enhanced cell proliferation, invasion, and migration in vitro. Stable overexpression of miR-182 also facilitated tumor growth and metastasis in vivo too. Further research showed that miR-182 could directly target the 3'untranslated region (3'UTR) of SATB2 mRNA and subsequently repress both the mRNA and protein expressions of SATB2, which we identified in previous studies as a CRC metastasis-associated protein. Restoring SATB2 expression could reverse the effects of miR-182 on CRC cell proliferation and migration. Investigations of possible mechanisms underlying these behaviors induced by miR-182 revealed that miR-182 induced epithelial-mesenchymal transition (EMT) by modulating the expression of key cellular molecules in EMT.

Conclusions: Our results illustrated that the up-regulation of miR-182 played a pivotal role in CRC tumorigenesis and metastasis, which suggesting a potential implication of miR-182 in the molecular therapy for CRC.

Show MeSH
Related in: MedlinePlus