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MicroRNA-493 suppresses tumor growth, invasion and metastasis of lung cancer by regulating E2F1.

Gu Y, Cheng Y, Song Y, Zhang Z, Deng M, Wang C, Zheng G, He Z - PLoS ONE (2014)

Bottom Line: Our data indicated that the expression of miR-493 was markedly reduced in pulmonary carcinoma.The ectopic expression of miR-493 impaired cell growth and invasion in vitro and in vivo.Mechanically, miR-493 commonly directly targeted E2F1, which resulted in a robust reduction of the expression of mRNA and protein.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute and Cancer Hospital, Guangzhou Medical University, Guangzhou, Guangdong, PR China; Medical School, University of South China, Hengyang, Hunan, PR China.

ABSTRACT
miRNAs have been proposed to be key regulators of progression and metastasis in cancer. However, an understanding of their roles and molecular mechanisms is needed to provide deeper insights for better therapeutic opportunities. In this study we investigated the role and mechanism of miR-493 in the development and progression of nonsmall-cell lung cancer (NSCLC). Our data indicated that the expression of miR-493 was markedly reduced in pulmonary carcinoma. The ectopic expression of miR-493 impaired cell growth and invasion in vitro and in vivo. Mechanically, miR-493 commonly directly targeted E2F1, which resulted in a robust reduction of the expression of mRNA and protein. This effect, in turn, decreased the growth, invasion and metastasis of lung cancer cells. Our findings highlight the importance of miR-493 dysfunction in promoting tumor progression, and implicate miR-493 as a potential therapeutic target in lung cancer.

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Expression of miR-493 inhibits the migration and invasion of lung cancer cells.A, scratch wound assays were conducted on 95D cells transfected with miR-493 and its paired control. The results from 3 separate assays were averaged together and graphed. *, P<0.05 (left). Representative images of the assays are shown. Original magnification: ×200 (right). B, the invasive properties of the H1975 and 95D cells were analyzed with a Transwell assay using a Matrigel-coated chamber. Migrated cells were plotted as the average number of cells per field of view from 3 different experiments, as described in the Materials and Methods. *P<0.01 (left). Representative images of the assays are shown. Original magnification: ×200 (right). C, in vivo metastasis assays were used to examine the lung metastatic ability of 95D/miR-493 cells labeled with green fluorescent protein(GFP) and its paired control cell 95D/control. Lung cancer cells were injected into the tail veins of five week- old mice. On day-28, all animals were sacrificed and the lungs were excised. The lung metastasis images were obtained with a Bruker Small Animal Imaging System. The lung tissue was then removed, fixed, paraffin-embedded, serially sectioned, and subjected to hematoxylin and eosin (H&E) staining. Left, Representation of the detected GFP signal in each of the five animals (the images were overlaid with green fluorescence and white light). Right, The metastatic field and fluorescence intensity significantly differed between the 95D/miR-493 group and the control group. D, Histological examination of pulmonary metastases from 95D/miR-493 and 95D/control cells by haematoxylin and eosin staining.
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pone-0102602-g003: Expression of miR-493 inhibits the migration and invasion of lung cancer cells.A, scratch wound assays were conducted on 95D cells transfected with miR-493 and its paired control. The results from 3 separate assays were averaged together and graphed. *, P<0.05 (left). Representative images of the assays are shown. Original magnification: ×200 (right). B, the invasive properties of the H1975 and 95D cells were analyzed with a Transwell assay using a Matrigel-coated chamber. Migrated cells were plotted as the average number of cells per field of view from 3 different experiments, as described in the Materials and Methods. *P<0.01 (left). Representative images of the assays are shown. Original magnification: ×200 (right). C, in vivo metastasis assays were used to examine the lung metastatic ability of 95D/miR-493 cells labeled with green fluorescent protein(GFP) and its paired control cell 95D/control. Lung cancer cells were injected into the tail veins of five week- old mice. On day-28, all animals were sacrificed and the lungs were excised. The lung metastasis images were obtained with a Bruker Small Animal Imaging System. The lung tissue was then removed, fixed, paraffin-embedded, serially sectioned, and subjected to hematoxylin and eosin (H&E) staining. Left, Representation of the detected GFP signal in each of the five animals (the images were overlaid with green fluorescence and white light). Right, The metastatic field and fluorescence intensity significantly differed between the 95D/miR-493 group and the control group. D, Histological examination of pulmonary metastases from 95D/miR-493 and 95D/control cells by haematoxylin and eosin staining.

Mentions: In addition to cell growth inhibition, the effect of miR-493 on tumor invasion and metastasis was also addressed in this study. A wound-healing assay showed that miR-493 could dramatically suppress tumor cell mobility in 95D cells compared with the empty vector-transfected cells (figure 3A). Furthermore, an invasion assay was also performed. The result demonstrated that miR-493 could suppress the invasive ability of lung cancer cells (figure 3B). An experimental animal model was utilized to further investigate the suppressive effect of miR-493 on tumor metastasis, 95D/miR-493 cells were injected into the tail veins of nude mice (five mice per group). Empty vector-transfected (95D/control) cells were used as controls. The mice were killed and the lungs were excised, and the metastatic lesions of the lung were then detected using a Bruker Small Animal Imaging System (Germany) after 28 days. The fields of metastatic lesions formation in the lung were significantly reduced compared to the control group (figure 3C). Although visible metastatic nodules were not observed on the surface of the lung, metastatic lesions were detected in the lung by haematoxylin and eosin staining (figure 4D). These results indicated that miR-493 could effectively suppress tumor metastasis in vitro and in vivo.


MicroRNA-493 suppresses tumor growth, invasion and metastasis of lung cancer by regulating E2F1.

Gu Y, Cheng Y, Song Y, Zhang Z, Deng M, Wang C, Zheng G, He Z - PLoS ONE (2014)

Expression of miR-493 inhibits the migration and invasion of lung cancer cells.A, scratch wound assays were conducted on 95D cells transfected with miR-493 and its paired control. The results from 3 separate assays were averaged together and graphed. *, P<0.05 (left). Representative images of the assays are shown. Original magnification: ×200 (right). B, the invasive properties of the H1975 and 95D cells were analyzed with a Transwell assay using a Matrigel-coated chamber. Migrated cells were plotted as the average number of cells per field of view from 3 different experiments, as described in the Materials and Methods. *P<0.01 (left). Representative images of the assays are shown. Original magnification: ×200 (right). C, in vivo metastasis assays were used to examine the lung metastatic ability of 95D/miR-493 cells labeled with green fluorescent protein(GFP) and its paired control cell 95D/control. Lung cancer cells were injected into the tail veins of five week- old mice. On day-28, all animals were sacrificed and the lungs were excised. The lung metastasis images were obtained with a Bruker Small Animal Imaging System. The lung tissue was then removed, fixed, paraffin-embedded, serially sectioned, and subjected to hematoxylin and eosin (H&E) staining. Left, Representation of the detected GFP signal in each of the five animals (the images were overlaid with green fluorescence and white light). Right, The metastatic field and fluorescence intensity significantly differed between the 95D/miR-493 group and the control group. D, Histological examination of pulmonary metastases from 95D/miR-493 and 95D/control cells by haematoxylin and eosin staining.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4126682&req=5

pone-0102602-g003: Expression of miR-493 inhibits the migration and invasion of lung cancer cells.A, scratch wound assays were conducted on 95D cells transfected with miR-493 and its paired control. The results from 3 separate assays were averaged together and graphed. *, P<0.05 (left). Representative images of the assays are shown. Original magnification: ×200 (right). B, the invasive properties of the H1975 and 95D cells were analyzed with a Transwell assay using a Matrigel-coated chamber. Migrated cells were plotted as the average number of cells per field of view from 3 different experiments, as described in the Materials and Methods. *P<0.01 (left). Representative images of the assays are shown. Original magnification: ×200 (right). C, in vivo metastasis assays were used to examine the lung metastatic ability of 95D/miR-493 cells labeled with green fluorescent protein(GFP) and its paired control cell 95D/control. Lung cancer cells were injected into the tail veins of five week- old mice. On day-28, all animals were sacrificed and the lungs were excised. The lung metastasis images were obtained with a Bruker Small Animal Imaging System. The lung tissue was then removed, fixed, paraffin-embedded, serially sectioned, and subjected to hematoxylin and eosin (H&E) staining. Left, Representation of the detected GFP signal in each of the five animals (the images were overlaid with green fluorescence and white light). Right, The metastatic field and fluorescence intensity significantly differed between the 95D/miR-493 group and the control group. D, Histological examination of pulmonary metastases from 95D/miR-493 and 95D/control cells by haematoxylin and eosin staining.
Mentions: In addition to cell growth inhibition, the effect of miR-493 on tumor invasion and metastasis was also addressed in this study. A wound-healing assay showed that miR-493 could dramatically suppress tumor cell mobility in 95D cells compared with the empty vector-transfected cells (figure 3A). Furthermore, an invasion assay was also performed. The result demonstrated that miR-493 could suppress the invasive ability of lung cancer cells (figure 3B). An experimental animal model was utilized to further investigate the suppressive effect of miR-493 on tumor metastasis, 95D/miR-493 cells were injected into the tail veins of nude mice (five mice per group). Empty vector-transfected (95D/control) cells were used as controls. The mice were killed and the lungs were excised, and the metastatic lesions of the lung were then detected using a Bruker Small Animal Imaging System (Germany) after 28 days. The fields of metastatic lesions formation in the lung were significantly reduced compared to the control group (figure 3C). Although visible metastatic nodules were not observed on the surface of the lung, metastatic lesions were detected in the lung by haematoxylin and eosin staining (figure 4D). These results indicated that miR-493 could effectively suppress tumor metastasis in vitro and in vivo.

Bottom Line: Our data indicated that the expression of miR-493 was markedly reduced in pulmonary carcinoma.The ectopic expression of miR-493 impaired cell growth and invasion in vitro and in vivo.Mechanically, miR-493 commonly directly targeted E2F1, which resulted in a robust reduction of the expression of mRNA and protein.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute and Cancer Hospital, Guangzhou Medical University, Guangzhou, Guangdong, PR China; Medical School, University of South China, Hengyang, Hunan, PR China.

ABSTRACT
miRNAs have been proposed to be key regulators of progression and metastasis in cancer. However, an understanding of their roles and molecular mechanisms is needed to provide deeper insights for better therapeutic opportunities. In this study we investigated the role and mechanism of miR-493 in the development and progression of nonsmall-cell lung cancer (NSCLC). Our data indicated that the expression of miR-493 was markedly reduced in pulmonary carcinoma. The ectopic expression of miR-493 impaired cell growth and invasion in vitro and in vivo. Mechanically, miR-493 commonly directly targeted E2F1, which resulted in a robust reduction of the expression of mRNA and protein. This effect, in turn, decreased the growth, invasion and metastasis of lung cancer cells. Our findings highlight the importance of miR-493 dysfunction in promoting tumor progression, and implicate miR-493 as a potential therapeutic target in lung cancer.

Show MeSH
Related in: MedlinePlus