Category ME L26 The Synergistic Effect of the PIK3CA gene mutation H1047R and the

Her 2 gene i

Abstract The PI3K signaling pathway is directly responsible for cell proliferation,

survival, and the many processes which contribute to the growth of

breast cancer. Phosphatidylinositol 3- kinases (PI3Ks) are a group of lipid

kinases that are major players in this pathway. Class I PI3Ks are

composed of two subunits, an 85 kDA adaptor subunit (p85) that lacks

PI3K activity, and a 110 kDA catalytic subunit (p110), which is coded by

the PIK3CA gene. PIK3CA has been found to have a 20-30% somatic

mutation rate in breast cancer. In recent studies, H1047R, a mutation

position of PIK3CA, was found to have the fastest and strongest growing

ability with or without the epidermal growth factor (EGF), and was the

most potent mutation. Her2 (c-erbB-2 / Neu), or the Human epidermal

growth factor 2, is a proto-oncogene that is over-expressed in 30% of

breast cancer, making Her 2 amplification the most frequent mutation in

breast cancer. Her 2 codes for a tyrosine-kinase growth factor receptor,

which regulates normal cell function, growth, and proliferation. In clinical

studies it has been shown that the PIK3CA gene is connected with Her 2,

maybe because the tyrosine kinase growth factor receptors that Her2

code for phosphorylate and activate the PI3K pathway. Furthermore in

clinical studies patients with no PIK3CA mutations had less resistance to

Her2 treatment than patients with any kind of PIK3CA mutation, indicating a

possible relationship between the two genes and also between the two

genes for treatment.

In this study, a cell proliferation, colony formation, and invasion assay

were performed using the MCF10A cell line with over-expressed H1047R

gene mutation, Her 2, both genes, or Lac Z as a control, to determine if the

two genes have a synergistic effect. Furthermore, the MCF10A cell line

with both over-expressed Her2 and H1047R were tested for cell growth in

the presence of different concentrations and combinations of the Her2

and PIK3CA inhibitors in order to see a possible benefit of having both

genes present during treatment with inhibitors.

In conclusion, it was found that the PIK3CA mutation H1047R and the

Her2 gene have a synergistic effect in the growth of breast cancer, since

in both the cell proliferation and colony formation assay the added effect

of both genes increased cell growth and colony formation numbers. Also,

when inhibitors were added to the cells with both Her2 and H1047R

mutations, it was proven that when the PIK3CA inhibitor LY294002 was

used when the Her2 inhibitor, CP-724714, was also added, cell growth

and colony formation was low also supporting their synergistic effect.

Overall, these results suggest that the mutation H1047R in the PIK3CA

gene and the Her2 gene are synergistic and that as a result these genes

may be a potent target for anti-cancer therapies in breast cancer.

Bibliography . Jitesh P. Jani, Richard S. Finn, Mary Campbell, et al. Discovery and

Pharmacologic Characterization of

CP-724,714, a Selective ErbB2 Tyrosine Kinase Inhibitor Cancer Res

2007;67:9887-9893.
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