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Overexpression of microRNA miR-30a or miR-191 in A549 lung cancer or BEAS-2B normal lung cell lines does not alter phenotype.

Patnaik SK, Kannisto E, Yendamuri S - PLoS ONE (2010)

Bottom Line: When compared to the corresponding controls, both cell lines overexpressing miR-30a or -191 do not demonstrate any significant changes in cell cycle distribution, cell proliferation, adherent colony formation, soft agar colony formation, xenograft formation in a subcutaneous SCID mouse model, and drug sensitivity to doxorubicin and cisplatin.There is a modest increase in cell migration in cell lines overexpressing miR-30a compared to their controls.Overexpression of miR-30a or -191 does not lead to an alteration in cell cycle, proliferation, xenograft formation, and chemosensitivity of A549 and BEAS-2B cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Roswell Park Cancer Institute, Buffalo, New York, United States of America.

ABSTRACT

Background: MicroRNAs (miRNAs) are small, noncoding RNAs (ribonucleic acids) that regulate translation. Several miRNAs have been shown to be altered in whole cancer tissue compared to normal tissue when quantified by microarray. Based on previous such evidence of differential expression, we chose to study the functional significance of miRNAs miR-30a and -191 alterations in human lung cancer.

Methodology/principal findings: The functional significance of miRNAs miR-30a and -191 was studied by creating stable transfectants of the lung adenocarcinoma cell line A549 and the immortalized bronchial epithelial cell line BEAS-2B with modest overexpression of miR-30a or -191 using a lentiviral system. When compared to the corresponding controls, both cell lines overexpressing miR-30a or -191 do not demonstrate any significant changes in cell cycle distribution, cell proliferation, adherent colony formation, soft agar colony formation, xenograft formation in a subcutaneous SCID mouse model, and drug sensitivity to doxorubicin and cisplatin. There is a modest increase in cell migration in cell lines overexpressing miR-30a compared to their controls.

Conclusions/significance: Overexpression of miR-30a or -191 does not lead to an alteration in cell cycle, proliferation, xenograft formation, and chemosensitivity of A549 and BEAS-2B cell lines. Using microarray data from whole tumors to select specific miRNAs for functional study may be a suboptimal strategy.

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Related in: MedlinePlus

Quantitative RT-PCR for confirming over-expression of miR-30a in A-30a and B-30a, and of miR-191 in A-191 and B-191 cell lines, respectively.Expression is normalized to control cell lines (A0 or B0).
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pone-0009219-g001: Quantitative RT-PCR for confirming over-expression of miR-30a in A-30a and B-30a, and of miR-191 in A-191 and B-191 cell lines, respectively.Expression is normalized to control cell lines (A0 or B0).

Mentions: Stable cell lines overexpressing miR-30a and -191 were generated using a lentiviral system as described above. MiR-30a was overexpressed ∼2 fold and ∼3 fold in both A549 (A-30a) and BEAS-2B (B-30a) respectively compared to controls (A0 and B0 respectively). Compared to controls, miR-191 was overexpressed ∼2 fold and ∼7 fold in A549 (A-191) and BEAS-2B (B-191) respectively (Figure 1). Overexpression was also confirmed by visualization of RFP containing cells by microscopy.


Overexpression of microRNA miR-30a or miR-191 in A549 lung cancer or BEAS-2B normal lung cell lines does not alter phenotype.

Patnaik SK, Kannisto E, Yendamuri S - PLoS ONE (2010)

Quantitative RT-PCR for confirming over-expression of miR-30a in A-30a and B-30a, and of miR-191 in A-191 and B-191 cell lines, respectively.Expression is normalized to control cell lines (A0 or B0).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2821397&req=5

pone-0009219-g001: Quantitative RT-PCR for confirming over-expression of miR-30a in A-30a and B-30a, and of miR-191 in A-191 and B-191 cell lines, respectively.Expression is normalized to control cell lines (A0 or B0).
Mentions: Stable cell lines overexpressing miR-30a and -191 were generated using a lentiviral system as described above. MiR-30a was overexpressed ∼2 fold and ∼3 fold in both A549 (A-30a) and BEAS-2B (B-30a) respectively compared to controls (A0 and B0 respectively). Compared to controls, miR-191 was overexpressed ∼2 fold and ∼7 fold in A549 (A-191) and BEAS-2B (B-191) respectively (Figure 1). Overexpression was also confirmed by visualization of RFP containing cells by microscopy.

Bottom Line: When compared to the corresponding controls, both cell lines overexpressing miR-30a or -191 do not demonstrate any significant changes in cell cycle distribution, cell proliferation, adherent colony formation, soft agar colony formation, xenograft formation in a subcutaneous SCID mouse model, and drug sensitivity to doxorubicin and cisplatin.There is a modest increase in cell migration in cell lines overexpressing miR-30a compared to their controls.Overexpression of miR-30a or -191 does not lead to an alteration in cell cycle, proliferation, xenograft formation, and chemosensitivity of A549 and BEAS-2B cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Thoracic Surgery, Roswell Park Cancer Institute, Buffalo, New York, United States of America.

ABSTRACT

Background: MicroRNAs (miRNAs) are small, noncoding RNAs (ribonucleic acids) that regulate translation. Several miRNAs have been shown to be altered in whole cancer tissue compared to normal tissue when quantified by microarray. Based on previous such evidence of differential expression, we chose to study the functional significance of miRNAs miR-30a and -191 alterations in human lung cancer.

Methodology/principal findings: The functional significance of miRNAs miR-30a and -191 was studied by creating stable transfectants of the lung adenocarcinoma cell line A549 and the immortalized bronchial epithelial cell line BEAS-2B with modest overexpression of miR-30a or -191 using a lentiviral system. When compared to the corresponding controls, both cell lines overexpressing miR-30a or -191 do not demonstrate any significant changes in cell cycle distribution, cell proliferation, adherent colony formation, soft agar colony formation, xenograft formation in a subcutaneous SCID mouse model, and drug sensitivity to doxorubicin and cisplatin. There is a modest increase in cell migration in cell lines overexpressing miR-30a compared to their controls.

Conclusions/significance: Overexpression of miR-30a or -191 does not lead to an alteration in cell cycle, proliferation, xenograft formation, and chemosensitivity of A549 and BEAS-2B cell lines. Using microarray data from whole tumors to select specific miRNAs for functional study may be a suboptimal strategy.

Show MeSH
Related in: MedlinePlus