A total of 107 genes were significantly regulated by hypoxia, 28 genes were up regulated and 79 genes were down scientific research regulated. Hypoxia expression patterns differed between histological subtypes. Four genes were significantly regulated in the same direction in both subtypes with a minimal two fold change, PPP1R3C, KCTD11, FAM115C, and membrane metallo endopeptidase. The GO annotations for the gene products are as follows, PPP1R3C, regulation of glycogen biosynthesis, KCTD11, regulation of cell proliferation, and MME, proteolysis. The gene product of FAM115C has unknown function. Hypoxia regulation of the four overlapping hypoxia genes and of the known hypoxia responsive gene hexokinase 2 was confirmed using real time PCR in normoxic and hypoxic fragments from an independent validation set.

Interestingly, the overall impact of hypoxia on gene expression was lower than Inhibitors,Modulators,Libraries the impact of histology or inter patient variability. Normoxic and hypoxic fragments derived from each pa tient clustered together significantly in 9 of 10 patients in pvclust Inhibitors,Modulators,Libraries analysis. Both clusters on the top of the hierarchy were significant in pvclust analysis. One cluster contained four squamous cell carcinomas, Inhibitors,Modulators,Libraries the other cluster contained all adenocarcin omas and one squamous cell carcinoma. MME immune histochemistry In order to determine the cell types responsible for MME expression in our model we performed immuno histochemical staining in fresh NSCLC specimens from 12 patients. MME positive neoplastic tumor cells were found in 80% and scattered MME positive stroma cells were found in 54% of fresh cancer specimens.

Up to 30% of stroma cells were MME positive Inhibitors,Modulators,Libraries in cultured frag ments, indicating generally increased MME expression in tumor stroma cells under stress conditions. Using this technique, no difference in MME staining in normoxia or hypoxia was found. However, since immuno histochemistry is a semiquantitative method, only large differences in expression levels can be detected. Next, consecutive sections of fresh NSCLC samples from 30 pa tients were stained for MME and HIF 1 in order to analyze, whether Inhibitors,Modulators,Libraries the expression of both is linked in vivo. Similar to the first series MME staining was found in tumor cells in 21 30 samples and in stroma cells in 10 30 samples. In 8 30 patients, HIF 1 positivity was found in tumor cells. In 2 30 patients also stroma cells were HIF 1 positive.

In a sample with very high stroma and tumor cell HIF 1 expression, HIF 1 and MME staining overlapped in stroma cells, but not in tumor cells. On the other hand in another patient with MME stroma staining no HIF 1 was found. In tumor cells MME and HIF 1 staining Ponatinib were not strongly related. Together this indicated to us that in some patients hypoxia may be linked to MME expres sion in the tumor stroma.