Limits...
Impedimetric detection of mutant p53 biomarker-driven metastatic breast cancers under hyposmotic pressure.

Shi M, Shtraizent N, Polotskaia A, Bargonetti J, Matsui H - PLoS ONE (2014)

Bottom Line: The results showed that knockdown of mtp53 leads to decrease in cell swelling.In addition, by means of two types of impedimetric detection systems we consistently detected enhancement of impedance signal in mtp53-expressing breast cancer cells.The identification via the electric measurement can be accomplished within 10 minutes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Hunter College and the Graduate Center, City University of New York, New York, New York, United States of America.

ABSTRACT
In cancer cells, the oncogenic mutant p53 (mtp53) protein is present at high levels and gain-of-function (GOF) activities with more expression of mtp53 proteins contribute to tumor growth and metastasis. Robust analytical approaches that probe the degree of metastasis of cancer cells in connection with the mtp53 activity will be extremely useful not only for establishing a better cancer prognosis but also understanding the fundamental mechanism of mtp53 oncogenic action. Here we assessed the influence of mtp53 in breast cancers to the mechanical property of breast cancer cells. Recently, ovarian and kidney cancer cell lines have been shown to have higher cellular elasticity as compared to normal cells assessed by monitoring the degree of deformation under hyposmotic pressure. To make fast detection in large scale, the impedance measurement was applied to monitor the swelling ratio of cells with time. The results showed that knockdown of mtp53 leads to decrease in cell swelling. In addition, by means of two types of impedimetric detection systems we consistently detected enhancement of impedance signal in mtp53-expressing breast cancer cells. Based on this observation we hypothesize that highly expressed mtp53 in metastatic mutant breast cancers can promote tumor progression by making cells more deformable and easier to spread out through extracellular matrix. The identification via the electric measurement can be accomplished within 10 minutes. All results in this report suggest that electric probing for the extent of the mtp53 expression of breast cancer cells may serve as a meaningful fingerprint for the cancer diagnostics, and this outcome will also have an important clinical implication for the development of mtp53-based targeting for tumor detection and treatment.

Show MeSH

Related in: MedlinePlus

Schematic illustration of the response of various breast cancer cell lines to doxycycline treatment.The degree of the expression of mtp53 in shp53 cell lines (Figures 1 A–C) can be controlled with the treatment of dox, while the mtp53 expression in parental and vector-STGM cell lines remains unchanged.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4060997&req=5

pone-0099351-g001: Schematic illustration of the response of various breast cancer cell lines to doxycycline treatment.The degree of the expression of mtp53 in shp53 cell lines (Figures 1 A–C) can be controlled with the treatment of dox, while the mtp53 expression in parental and vector-STGM cell lines remains unchanged.

Mentions: The aim of our work is to probe changes in the mechanoelastic property of breast cancer cells associated with the expression of mtp53. We hypothesized that high expression of mtp53 in two types of breast cancer cells (MDA-MB-468 (with mtp53 R280H) and MDA-MB-231 (with mtp53 R273L)) leads to softening of cell structure and the hyposmotic pressure induces swelling of these cells in contrast with ones devoid of mtp53. Thus we expected that the removal of mtp53 would decrease elasticity and revert cells to a stiffer normal phenotype. To test this hypothesis, we induced depletion of mtp53 in shRNA-mediated knockdown cell lines, MDA-231.shp53 (Figure 1-A, B) and MDA-468.shp53 (Figure 1-C), and analyzed their mechanoelastic change before and after the knockdown. The shRNA cancer cells were generated through miR30-based knockdown of mtp53 induced by doxycycline. While doxycycline controls the degree of the expression of mtp53 in shp53 cell lines (Figures 1-A–C) it does not influence the mtp53 expression in parental and vector-STGM cell lines (Figures 1-D–F); The engineered MDA-MB-231 vector STGM cells and MDA-MB-468 vector STGM cells express the same proteins as MDA-MB-231 and MDA-MB-468 except without mtp53.shRNA and thus doxycycline cannot knockdown mtp53. Therefore, the parental MDA-MB-231, MDA-MB-468 vector STGM and MDA-MB-231 vector STGM cells served as negative controls. And the cellular mechanical property should also reflect this metastatic behavior change based on our hypothesis.


Impedimetric detection of mutant p53 biomarker-driven metastatic breast cancers under hyposmotic pressure.

Shi M, Shtraizent N, Polotskaia A, Bargonetti J, Matsui H - PLoS ONE (2014)

Schematic illustration of the response of various breast cancer cell lines to doxycycline treatment.The degree of the expression of mtp53 in shp53 cell lines (Figures 1 A–C) can be controlled with the treatment of dox, while the mtp53 expression in parental and vector-STGM cell lines remains unchanged.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4060997&req=5

pone-0099351-g001: Schematic illustration of the response of various breast cancer cell lines to doxycycline treatment.The degree of the expression of mtp53 in shp53 cell lines (Figures 1 A–C) can be controlled with the treatment of dox, while the mtp53 expression in parental and vector-STGM cell lines remains unchanged.
Mentions: The aim of our work is to probe changes in the mechanoelastic property of breast cancer cells associated with the expression of mtp53. We hypothesized that high expression of mtp53 in two types of breast cancer cells (MDA-MB-468 (with mtp53 R280H) and MDA-MB-231 (with mtp53 R273L)) leads to softening of cell structure and the hyposmotic pressure induces swelling of these cells in contrast with ones devoid of mtp53. Thus we expected that the removal of mtp53 would decrease elasticity and revert cells to a stiffer normal phenotype. To test this hypothesis, we induced depletion of mtp53 in shRNA-mediated knockdown cell lines, MDA-231.shp53 (Figure 1-A, B) and MDA-468.shp53 (Figure 1-C), and analyzed their mechanoelastic change before and after the knockdown. The shRNA cancer cells were generated through miR30-based knockdown of mtp53 induced by doxycycline. While doxycycline controls the degree of the expression of mtp53 in shp53 cell lines (Figures 1-A–C) it does not influence the mtp53 expression in parental and vector-STGM cell lines (Figures 1-D–F); The engineered MDA-MB-231 vector STGM cells and MDA-MB-468 vector STGM cells express the same proteins as MDA-MB-231 and MDA-MB-468 except without mtp53.shRNA and thus doxycycline cannot knockdown mtp53. Therefore, the parental MDA-MB-231, MDA-MB-468 vector STGM and MDA-MB-231 vector STGM cells served as negative controls. And the cellular mechanical property should also reflect this metastatic behavior change based on our hypothesis.

Bottom Line: The results showed that knockdown of mtp53 leads to decrease in cell swelling.In addition, by means of two types of impedimetric detection systems we consistently detected enhancement of impedance signal in mtp53-expressing breast cancer cells.The identification via the electric measurement can be accomplished within 10 minutes.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Hunter College and the Graduate Center, City University of New York, New York, New York, United States of America.

ABSTRACT
In cancer cells, the oncogenic mutant p53 (mtp53) protein is present at high levels and gain-of-function (GOF) activities with more expression of mtp53 proteins contribute to tumor growth and metastasis. Robust analytical approaches that probe the degree of metastasis of cancer cells in connection with the mtp53 activity will be extremely useful not only for establishing a better cancer prognosis but also understanding the fundamental mechanism of mtp53 oncogenic action. Here we assessed the influence of mtp53 in breast cancers to the mechanical property of breast cancer cells. Recently, ovarian and kidney cancer cell lines have been shown to have higher cellular elasticity as compared to normal cells assessed by monitoring the degree of deformation under hyposmotic pressure. To make fast detection in large scale, the impedance measurement was applied to monitor the swelling ratio of cells with time. The results showed that knockdown of mtp53 leads to decrease in cell swelling. In addition, by means of two types of impedimetric detection systems we consistently detected enhancement of impedance signal in mtp53-expressing breast cancer cells. Based on this observation we hypothesize that highly expressed mtp53 in metastatic mutant breast cancers can promote tumor progression by making cells more deformable and easier to spread out through extracellular matrix. The identification via the electric measurement can be accomplished within 10 minutes. All results in this report suggest that electric probing for the extent of the mtp53 expression of breast cancer cells may serve as a meaningful fingerprint for the cancer diagnostics, and this outcome will also have an important clinical implication for the development of mtp53-based targeting for tumor detection and treatment.

Show MeSH
Related in: MedlinePlus