The slides had been counter stained with hematoxylin plus the photographs were captured which has a digital cam era. The signals were then measured utilizing ImageScope Software. Positivity was quantified by specifying a hue value of 0. one and hue width of 0. 33 for any standardized area of the tumor tissue. Approval for this research was obtained through the Institu tional Ethics Assessment Board with the Scott White Me morial Hospital and Texas A &M Health Science Center. The research was conducted in compliance with the Helsinki Declaration. Statistics The co existence of Mcl one and either Bcl xL or USP9X expression in tumor cells were assessed making use of the Chi square and Fisher exact tests. A Pearson correlation be tween the Bcl xL, USP9X and Mcl one expression profiles was calculated making use of R statistical program. The association of protein expression and clinical staging within the tumor samples was determined implementing linear regression.
Results Mcl one and Bcl xL are co overexpressed in multiple solid tumor types To evaluate the correlation between Bcl xL and Mcl one expression in lung and colon cancer, we analyzed human non small cell lung adenocarcinoma and colon adeno carcinoma samples by immunohistochemistry working with antibodies against these two proteins. As shown in Figure 1a, there had been reversible FAK inhibitor strong associations observed be tween the expression of Mcl one and that of Bcl xL in both the lung and colon cancer samples. In the 117 human colon cancer samples we analyzed, 47 specimens stained positively for both proteins and a further 29 samples showed weak co staining for both factors. In the 81 lung cancer samples tested in this analysis, 51 samples showed strong positive staining for both proteins and five samples showed co staining at low levels. There were further relationships observed be tween Mcl one and Bcl xL protein PLX4720 expression and tumor staging in colon cancer samples.
Mcl 1 ex pression was found to increase with the staging grade,. Bcl xL expression was also found to be significantly associated with staging, with stage

I lesions showing significantly dif ferent levels of this protein compared with stage III and stage IV tumors. Tumor sta ging data have been not available for the lung cancer samples. Tumor cells expressing high levels of Mcl one and Bcl xL protein exhibit chemoresistance To test the hypothesis that high Mcl one and Bcl xL expression contributes to drug resistance, including re sistance to Bcl xL inhibitors, the baseline protein expres sions of Bcl xL and Mcl 1 in multiple cell lines have been examined via western blotting. The results demonstrated the concurrent expression of both Mcl 1 and Bcl xL in most cell lines, corroborating the immu nostaining results in both lung and colon tumor tissues shown in Figure 1.