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MircoRNA-33a inhibits epithelial-to-mesenchymal transition and metastasis and could be a prognostic marker in non-small cell lung cancer.

Yang L, Yang J, Li J, Shen X, Le Y, Zhou C, Wang S, Zhang S, Xu D, Gong Z - Sci Rep (2015)

Bottom Line: Here we found that miR-33a, an intronic miRNA located within the sterol regulatory element-binding protein 2 (SREBP-2) gene, is expressed at low levels in metastatic non-small cell lung cancer (NSCLC) cells and is inversely correlated with Twist1 expression.Additionally, Twist1 knockdown blocks EMT-related metastasis and forced expression of miR-33a inhibits lung cancer metastasis in a xenograft animal model.Clinically, miR-33a is found to be at low levels in NSCLC patients and down-regulation of miR-33a predicts a poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Biology.

ABSTRACT
Understanding the molecular mechanism by which epithelial mesenchymal transition (EMT)-mediated cancer metastasis and how microRNA (miRNA) regulates lung cancer progression via Twist1-activated EMT may provide potential therapeutic targets for cancer therapy. Here we found that miR-33a, an intronic miRNA located within the sterol regulatory element-binding protein 2 (SREBP-2) gene, is expressed at low levels in metastatic non-small cell lung cancer (NSCLC) cells and is inversely correlated with Twist1 expression. Conversely, miR-33a knockdown induces EMT and miR-33a overexpression blocks EMT by regulating of Twist1 expression in NSCLC cells. Bioinformatical prediction and luciferase reporter assay confirm that Twist1 is a direct target of miR-33a. Additionally, Twist1 knockdown blocks EMT-related metastasis and forced expression of miR-33a inhibits lung cancer metastasis in a xenograft animal model. Clinically, miR-33a is found to be at low levels in NSCLC patients and down-regulation of miR-33a predicts a poor prognosis. These findings suggest that miR-33a targets Twist1 and inhibits invasion and metastasis in NSCLC. Thus, miR-33a might be a potential prognostic marker and of therapeutic relevance for NSCLC metastasis intervention.

No MeSH data available.


Related in: MedlinePlus

miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis.(A) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and (B) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 (C) and NCI-H1299 (D) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 (E,G) and NCI-H1299 (F,H) cells. *P < 0.05, **P < 0.01, ***P < 0.001.
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f2: miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis.(A) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and (B) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 (C) and NCI-H1299 (D) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 (E,G) and NCI-H1299 (F,H) cells. *P < 0.05, **P < 0.01, ***P < 0.001.

Mentions: To determine whether miR-33a is directly involved in EMT-dependent metastasis, the effect of miR-33a on cell EMT property was examined. The expression of miR-33a was down-regulated by the transfection of miR-33a inhibitors into the low-metastasis SPC-A-1 cell line, which expresses a relatively high level of endogenous miR-33a (Fig. 2A). In the high-metastasis NCI-H1299 cell line, the transfection of a miR-33a mimic resulted in miR-33a overexpression compared with the control RNA treatment (Fig. 2B). The results of Western blotting showed that the knockdown of miR-33a induced a marked decrease of the epithelial cell biomarker E-cadherin (CDH1) and an increase of the mesenchymal cell biomarker Vimentin (Fig. 2C), which indicated that cells had undergone an EMT. Additionally, miR-33a knockdown promoted cell migration (Fig. 2E) and invasion (Fig. 2G) in SPC-A-1 cells. In contrast, miR-33a replacement in NCI-H1299 cells increased the expression of the epithelial cell biomarker of CDH1 and decreased the expression of the mesenchymal cell biomarker Vimentin (Fig. 2D), which indicated a block of EMT. In addition, miR-33a overexpression reduced the migration (Fig. 2F) and invasion (Fig. 2H) capability of NCI-H1299 cells. Thus, the data show that miR-33a is a crucial mediator of EMT and metastasis in NSCLC cells.


MircoRNA-33a inhibits epithelial-to-mesenchymal transition and metastasis and could be a prognostic marker in non-small cell lung cancer.

Yang L, Yang J, Li J, Shen X, Le Y, Zhou C, Wang S, Zhang S, Xu D, Gong Z - Sci Rep (2015)

miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis.(A) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and (B) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 (C) and NCI-H1299 (D) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 (E,G) and NCI-H1299 (F,H) cells. *P < 0.05, **P < 0.01, ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f2: miR-33a knockdown induces EMT and metastasis and miR-33a overexpression blocks EMT and metastasis.(A) SPC-A-1 cells was transfected with miR-33a inhibitors (anti-miR-33a) or inhibitor NC (Ctrl RNA) and (B) NCI-H1299 cells was transfected with miR-33a mimics (miR-33a) or mimics NC (Ctrl RNA), the levels of mature miR-33a were confirmed by quantitative RT-PCR. Western Blotting was performed after the transfection of RNAs. The changes of epithelial cell biomarker E-cadherin and mesenchymal cell biomarker Vimentin were shown in SPC-A-1 (C) and NCI-H1299 (D) cells. The migration and invasion capability were respectively measured after the transfection of RNA oligos in SPC-A-1 (E,G) and NCI-H1299 (F,H) cells. *P < 0.05, **P < 0.01, ***P < 0.001.
Mentions: To determine whether miR-33a is directly involved in EMT-dependent metastasis, the effect of miR-33a on cell EMT property was examined. The expression of miR-33a was down-regulated by the transfection of miR-33a inhibitors into the low-metastasis SPC-A-1 cell line, which expresses a relatively high level of endogenous miR-33a (Fig. 2A). In the high-metastasis NCI-H1299 cell line, the transfection of a miR-33a mimic resulted in miR-33a overexpression compared with the control RNA treatment (Fig. 2B). The results of Western blotting showed that the knockdown of miR-33a induced a marked decrease of the epithelial cell biomarker E-cadherin (CDH1) and an increase of the mesenchymal cell biomarker Vimentin (Fig. 2C), which indicated that cells had undergone an EMT. Additionally, miR-33a knockdown promoted cell migration (Fig. 2E) and invasion (Fig. 2G) in SPC-A-1 cells. In contrast, miR-33a replacement in NCI-H1299 cells increased the expression of the epithelial cell biomarker of CDH1 and decreased the expression of the mesenchymal cell biomarker Vimentin (Fig. 2D), which indicated a block of EMT. In addition, miR-33a overexpression reduced the migration (Fig. 2F) and invasion (Fig. 2H) capability of NCI-H1299 cells. Thus, the data show that miR-33a is a crucial mediator of EMT and metastasis in NSCLC cells.

Bottom Line: Here we found that miR-33a, an intronic miRNA located within the sterol regulatory element-binding protein 2 (SREBP-2) gene, is expressed at low levels in metastatic non-small cell lung cancer (NSCLC) cells and is inversely correlated with Twist1 expression.Additionally, Twist1 knockdown blocks EMT-related metastasis and forced expression of miR-33a inhibits lung cancer metastasis in a xenograft animal model.Clinically, miR-33a is found to be at low levels in NSCLC patients and down-regulation of miR-33a predicts a poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biochemistry and Molecular Biology.

ABSTRACT
Understanding the molecular mechanism by which epithelial mesenchymal transition (EMT)-mediated cancer metastasis and how microRNA (miRNA) regulates lung cancer progression via Twist1-activated EMT may provide potential therapeutic targets for cancer therapy. Here we found that miR-33a, an intronic miRNA located within the sterol regulatory element-binding protein 2 (SREBP-2) gene, is expressed at low levels in metastatic non-small cell lung cancer (NSCLC) cells and is inversely correlated with Twist1 expression. Conversely, miR-33a knockdown induces EMT and miR-33a overexpression blocks EMT by regulating of Twist1 expression in NSCLC cells. Bioinformatical prediction and luciferase reporter assay confirm that Twist1 is a direct target of miR-33a. Additionally, Twist1 knockdown blocks EMT-related metastasis and forced expression of miR-33a inhibits lung cancer metastasis in a xenograft animal model. Clinically, miR-33a is found to be at low levels in NSCLC patients and down-regulation of miR-33a predicts a poor prognosis. These findings suggest that miR-33a targets Twist1 and inhibits invasion and metastasis in NSCLC. Thus, miR-33a might be a potential prognostic marker and of therapeutic relevance for NSCLC metastasis intervention.

No MeSH data available.


Related in: MedlinePlus