Data on the putative role of p53 in breast cancer cell adhesion: Technical information for adhesion assay

In this data article, the potential role of p53 tumor suppressor gene (p53) on the attachment ability of MCF-7 breast cancer cells was investigated. In our main article, “IGF-I/ EGF and E2 signaling crosstalk through IGF-IR conduit point affect breast cancer cell adhesion” (K. Voudouri, D. Nikitovic, A. Berdiaki, D. Kletsas, N.K. Karamanos, G.N. Tzanakakis, 2016) [1], we describe the key role of IGF-IR in breast cancer cell adhesion onto fibronectin (FN). p53 tumor suppressor gene is a principal regulator of cancer cell proliferation. Various data have demonstrated an association between p53 and IGF-IR actions on cell growth through its’ putative regulation of IGF-IR expression. According to our performed experiments, p53 does not modify IGF-IR expression and does not affect basal MCF-7 cells adhesion onto FN. Moreover, technical details about the performance of adhesion assay onto the FN substrate were provided.


a b s t r a c t
In this data article, the potential role of p53 tumor suppressor gene (p53) on the attachment ability of MCF-7 breast cancer cells was investigated. In our main article, "IGF-I/ EGF and E2 signaling crosstalk through IGF-IR conduit point affect breast cancer cell adhesion" (K. Voudouri, D. Nikitovic, A. Berdiaki, D. Kletsas, N.K. Karamanos, G.N. Tzanakakis, 2016) [1], we describe the key role of IGF-IR in breast cancer cell adhesion onto fibronectin (FN). p53 tumor suppressor gene is a principal regulator of cancer cell proliferation. Various data have demonstrated an association between p53 and IGF-IR actions on cell growth through its' putative regulation of IGF-IR expression. According to our performed experiments, p53 does not modify IGF-IR expression and does not affect basal MCF-7 cells adhesion onto FN. Moreover Various numbers of cells were plated onto FN to optimize cell number and adhesion time in order to design a tailor-made adhesion assay for MCF-7 cells.
The attachment ability of cells and the expression of IGF-IR were assessed after transfection of cells with siRNA specific for p53 gene.

Data source location
Department of Anatomy-Histology-Embryology, School of Medicine, University of Crete

Data accessibility
Data are provided with this article

Value of the data
There is an established connection between p53 and IGF-IR activities in cancer; specifically p53 mutant forms are known to enhance IGF-IR gene expression (2)(3)(4)(5). The assessment of p53/IGF-IR interactions in breast cancer cell adhesion can be of value for research groups from related fields.
These data can be compared to other scientific data addressing the connection between various tumor suppressor genes and IGF-IR expression and/or to data of this interaction in other cell lines and functions.
These data facilitates other researchers to execute the optimum adhesion assay for the evaluation of MCF-7 cell adhesion onto FN.

Data
This article contains graphs presenting data on the role of p53 tumor-suppressor gene [2][3][4][5] in IGF-IR expression and breast cancer cell adhesion (Fig. 1). Furthermore, technical details for the performance of the MCF-7 cells' adhesion assay including number of plated cells and the adherence time for the MCF-7 cell adhesion protocol, are included (Fig. 2). Utilized reagents are presented in Table 1.

Experimental design, materials and methods
In order to optimize the adhesion assay protocol we utilized various cell seeding numbers and adhesion times. Cell lines and cell culture conditions are presented in [1]. In this article additional technical features of cell adhesion assay are provided.

Real time PCR, Western blot, adhesion assay
Adhesion assay, siRNA transfection with siRNA specific for p53, real time PCR and western blot experiments were designed to examine the role of p53 in MCF-7 FN-dependent adhesion. RNA interference, Western Blot, Real time PCR and adhesion assay protocols were described in [1]. Here, we provide extra information describing the characteristics of utilized siRNA specific for p53, p53 antibody and p53 primer (Table 1).

Cell adherence assay-optimization of cell number and time to adhere
The 96-well plates were coated with FN (Milipore) (5 μg/ml) as described in (1). MCF-7 cells were detached with 5 mM PBSEDTA. Cells at 3000, 6000 and 9000 cell/well were seeded onto FN coated 96-well plate. In this experiment, the cells were allowed to adhere or 30 min. The 6000 cells/well approach was chosen as having the optimum seeded cells number / adhered cell number ratio. To determine the optimum adhesion time the 6000 cells/well were allowed to adhere during 30 min, 1 h, 1,5 h, 2 h and 3 h, respectively. The number of adherent cells was determined as described in (1) Table 1 Biochemical reagents utilized for the p53 experiment.

Transparency document. Supporting information
Transparency data associated with this article can be found in the online version at http://dx.doi. org/10.1016/ j.dib.2016.09.038.