te down regulation of Mcl 1. Notably, e pression levels of Mcl 1 in the three cell lines was high compared to that found in the non transformed mammary epithelial cell line MCF10A, indicating that signaling pathways leading to enhanced e pression Tofacitinib Citrate side effects of Mcl 1 are active in transformed mammary epithelial cells, and in HER2 overe pressing ones in particular. Transformed mammary epithelial cells, including established breast cancer cell lines such as BT474 cells, e hibit an inherent phenotypic plasticity and har bor a subpopulation of cells with features of cancer initiating cells. The latter cells, which are charac terized by numerous parameters, including their ability to form spherical colonies in non adherent culture con ditions, were frequently described as being resistant to cell death induction by numerous sti muli.

Inhibitors,Modulators,Libraries This suggests Inhibitors,Modulators,Libraries that they may rely on survival signals distinct from these that are critical for the rest of the population. We thus investigated whether the Mcl 1 dependence of BT474 cells revealed above applies to the subpo pulation of CICs. To test this, we reasoned that, if BT474 CICs are Mcl 1 dependent, then a diminished ability to form mammospheres should be observed in a Inhibitors,Modulators,Libraries population of BT474 that has been depleted in Mcl 1. The ability of BT474 cells to form mammospheres after transfection with siRNAs was thus evaluated. As shown in Figure 2, the ability of Mcl 1 depleted BT474 cells to form mammospheres was significantly decreased compared to that of the Inhibitors,Modulators,Libraries same cells treated with a con trol siRNA.

In contrast, Bcl L or Bcl 2 knock down was insufficient by itself to affect mammosphere Carfilzomib for mation by BT474 cells. Taken together, these data indicate that the HER2 overe pressing BT474 cells require Mcl 1 to survive in vitro, and that this Mcl 1 dependence e tends to their subpopulation of CICs. To investigate whether pathways driving Mcl 1 e pres sion are specifically active in HER2 overe pressing can cers, compared to other breast cancers, we analyzed the e pression of 20 pro and anti apoptotic Bcl 2 family members from published gene e pression profiles of breast cancer patients. We based this analysis on studies in which the HER2 status of each tumor was available and had been evaluated by immunohistochemistry, and that were performed using Affymetri microar rays.

Two Erlotinib mechanism of action studies corresponded to these criteria, allowing to investigate e pression profiles of 41 HER2 overe pres sing tumors and 170 HER2 ones. Our evalua tion was performed in a probe matching way, using the 2 pooled aforementioned cohorts. Regarding the e pres sion of anti apoptotic genes, this evaluation revealed a statistically sig nificant enrichment, in HER2 overe pressing breast tumors compared to other breast tumors, in one MCL1 specific probe and also in one BCL2L1 one. In contrast, other breast tumors appeared sta tistically enriched for three BCL2 specific probes. Interestingly, when the evaluation was performed on a larger pool obtained by merging the two previous