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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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SATB2 was a tumor-suppressor in CRC and attenuated miR-182-mediated malignant phenotype in CRC. (A) Cell proliferation was measured by CCK-8 assay to compare the growth difference between SW480 cells with over-expression of STAB2 and SW480 mock cells with transfection of empty vector. (B, C) The invasiveness and migration ability were detected by matrigel invasion assay (B) and wound-healing assay (C) in SW480 cell after SATB2 over-expression. Cell proliferation (D), invasion (E) and migration (F) changes were detected between SW480/miR-182 cells and SW480/miR-182 cells with over-expression of STAB2. SATB2 could inhibit cell growth, invasion and migration of SW480/miR-182. SW480/miR-182 denoted CRC SW480 cells with stable over-expression of miR-182. * p < 0.05, ** p <0.001.
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Figure 6: SATB2 was a tumor-suppressor in CRC and attenuated miR-182-mediated malignant phenotype in CRC. (A) Cell proliferation was measured by CCK-8 assay to compare the growth difference between SW480 cells with over-expression of STAB2 and SW480 mock cells with transfection of empty vector. (B, C) The invasiveness and migration ability were detected by matrigel invasion assay (B) and wound-healing assay (C) in SW480 cell after SATB2 over-expression. Cell proliferation (D), invasion (E) and migration (F) changes were detected between SW480/miR-182 cells and SW480/miR-182 cells with over-expression of STAB2. SATB2 could inhibit cell growth, invasion and migration of SW480/miR-182. SW480/miR-182 denoted CRC SW480 cells with stable over-expression of miR-182. * p < 0.05, ** p <0.001.

Mentions: The loss of SATB2 mRNA or protein in CRC cells has been observed in our previous studies, but its role in metastasis is yet to be determined. We overexpressed SATB2 in SW480 cells by transiently transfecting with pCAG-SATB2 vector. Compared with SW480 mock cells that were transfected with empty vector, the ectopic over-expression of SATB2 induced obvious repressions of cell proliferation, cell mobility and invasion (p < 0.05, Figure 6A-C).


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)

SATB2 was a tumor-suppressor in CRC and attenuated miR-182-mediated malignant phenotype in CRC. (A) Cell proliferation was measured by CCK-8 assay to compare the growth difference between SW480 cells with over-expression of STAB2 and SW480 mock cells with transfection of empty vector. (B, C) The invasiveness and migration ability were detected by matrigel invasion assay (B) and wound-healing assay (C) in SW480 cell after SATB2 over-expression. Cell proliferation (D), invasion (E) and migration (F) changes were detected between SW480/miR-182 cells and SW480/miR-182 cells with over-expression of STAB2. SATB2 could inhibit cell growth, invasion and migration of SW480/miR-182. SW480/miR-182 denoted CRC SW480 cells with stable over-expression of miR-182. * p < 0.05, ** p <0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 6: SATB2 was a tumor-suppressor in CRC and attenuated miR-182-mediated malignant phenotype in CRC. (A) Cell proliferation was measured by CCK-8 assay to compare the growth difference between SW480 cells with over-expression of STAB2 and SW480 mock cells with transfection of empty vector. (B, C) The invasiveness and migration ability were detected by matrigel invasion assay (B) and wound-healing assay (C) in SW480 cell after SATB2 over-expression. Cell proliferation (D), invasion (E) and migration (F) changes were detected between SW480/miR-182 cells and SW480/miR-182 cells with over-expression of STAB2. SATB2 could inhibit cell growth, invasion and migration of SW480/miR-182. SW480/miR-182 denoted CRC SW480 cells with stable over-expression of miR-182. * p < 0.05, ** p <0.001.
Mentions: The loss of SATB2 mRNA or protein in CRC cells has been observed in our previous studies, but its role in metastasis is yet to be determined. We overexpressed SATB2 in SW480 cells by transiently transfecting with pCAG-SATB2 vector. Compared with SW480 mock cells that were transfected with empty vector, the ectopic over-expression of SATB2 induced obvious repressions of cell proliferation, cell mobility and invasion (p < 0.05, Figure 6A-C).

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