Drive the normal cell. With time the cell accumulates mutations and epigenetic changes, which alter the signaling and biochemical networks, and can lead to cell transformation and cancer [1]. Although there are a few cases in which a disease can be linked to one major signaling event (e.g. Bcr-Abl in CML [2]), in most tumors this is not the case. Genetic, epigenetic and environmental perturbations occur throughout tumor development. Usually, the tumor is dependent on several oncogenic signals. Furthermore, the intrinsic genomic instability of cancer cells leads to continual evolution and to intra-tumor heterogeneity [3]. The microarray technology has become a popular and common strategy to study gene regulation in cancer [4?]. Although gene expression can also be regulated at the level of DNA, by mutation or epigenetic modifications, as well as post-transcriptionally, mRNA levels are considered a legitimate measure of gene expression, and analysis of expression microarrays is a valid method for analysis of changes in cellular functions. There are several ways to analyze microarray data, as described in [8?0]. One of the main hurdles in microarray analysis is the heterogeneity between biological replicates. In most cases, the analyst attempts to smooth over the heterogeneity, and looks at averagedexpression changes that are significant in most or all of the replicates [11,12]. Cluster analysis then delineates groups with significant differences. Although for many purposes this average analysis is appropriate, heterogenic data reflect real differences between biological replicates. These differences, which are minimized when looking at average expression, can have profound phenotypic effects. In recent years, the concept of personalized therapy has gained popularity [13?5]. Two fundamental principles that underlie the concept of personalized cancer therapy are that significant genomic heterogeneity exists among tumors, even those derived from the same tissue of origin, and that these differences can play an important role in determining the likelihood of a clinical response to treatment with particular agents. Such genomic heterogeneity can involve differences in the spectrum of coding sequence mutations, 1081537 as well as focal gene amplifications, deletions, or translocations. It might also involve epigenetic changes in the expression profile of a tumor cell, although the sources of epigenetic variation among tumors remain poorly understood [16]. In this study, we have looked at tumor heterogeneity in mice of similar genetic background. These mice shared the same living conditions and were treated with the same carcinogens, and all developed squamous cell carcinoma. We compared the results of averaging microarray data with the results of analyzing each tumor on a case-by-case basis. The case-by-case analysis highlighted the surprising LED-209 biological activity degree of heterogeneity of oncogenic signaling between the mice.Heterogeneous Gene Expression in SCC DevelopmentMaterials and MethodsAs described by Quigley et al., male SPRET/Ei mice were mated with female FVB/N mice, and the female F1 hybrids were backcrossed to FVB/N males. Skin tumors were induced on dorsal back skin of the resulting FVBBX mice by treatment with dimethyl benzanthracene (DMBA) and tetradecanoyl-phorbol acetate (TPA). MedChemExpress Methionine enkephalin Multiple benign papillomas and malignant squamous cell carcinomas (SCC) developed. Normal tail skin, papillomas and carcinomas were harvested when mice were sacrificed due to pres.Drive the normal cell. With time the cell accumulates mutations and epigenetic changes, which alter the signaling and biochemical networks, and can lead to cell transformation and cancer [1]. Although there are a few cases in which a disease can be linked to one major signaling event (e.g. Bcr-Abl in CML [2]), in most tumors this is not the case. Genetic, epigenetic and environmental perturbations occur throughout tumor development. Usually, the tumor is dependent on several oncogenic signals. Furthermore, the intrinsic genomic instability of cancer cells leads to continual evolution and to intra-tumor heterogeneity [3]. The microarray technology has become a popular and common strategy to study gene regulation in cancer [4?]. Although gene expression can also be regulated at the level of DNA, by mutation or epigenetic modifications, as well as post-transcriptionally, mRNA levels are considered a legitimate measure of gene expression, and analysis of expression microarrays is a valid method for analysis of changes in cellular functions. There are several ways to analyze microarray data, as described in [8?0]. One of the main hurdles in microarray analysis is the heterogeneity between biological replicates. In most cases, the analyst attempts to smooth over the heterogeneity, and looks at averagedexpression changes that are significant in most or all of the replicates [11,12]. Cluster analysis then delineates groups with significant differences. Although for many purposes this average analysis is appropriate, heterogenic data reflect real differences between biological replicates. These differences, which are minimized when looking at average expression, can have profound phenotypic effects. In recent years, the concept of personalized therapy has gained popularity [13?5]. Two fundamental principles that underlie the concept of personalized cancer therapy are that significant genomic heterogeneity exists among tumors, even those derived from the same tissue of origin, and that these differences can play an important role in determining the likelihood of a clinical response to treatment with particular agents. Such genomic heterogeneity can involve differences in the spectrum of coding sequence mutations, 1081537 as well as focal gene amplifications, deletions, or translocations. It might also involve epigenetic changes in the expression profile of a tumor cell, although the sources of epigenetic variation among tumors remain poorly understood [16]. In this study, we have looked at tumor heterogeneity in mice of similar genetic background. These mice shared the same living conditions and were treated with the same carcinogens, and all developed squamous cell carcinoma. We compared the results of averaging microarray data with the results of analyzing each tumor on a case-by-case basis. The case-by-case analysis highlighted the surprising degree of heterogeneity of oncogenic signaling between the mice.Heterogeneous Gene Expression in SCC DevelopmentMaterials and MethodsAs described by Quigley et al., male SPRET/Ei mice were mated with female FVB/N mice, and the female F1 hybrids were backcrossed to FVB/N males. Skin tumors were induced on dorsal back skin of the resulting FVBBX mice by treatment with dimethyl benzanthracene (DMBA) and tetradecanoyl-phorbol acetate (TPA). Multiple benign papillomas and malignant squamous cell carcinomas (SCC) developed. Normal tail skin, papillomas and carcinomas were harvested when mice were sacrificed due to pres.