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Rachy, brachytherapy. Chemo, chemotherapy with Cisplatin. Status alive was registered at the last follow up, death was caused by primary tumor of cervical cancer, except the case labeled with an asterisk, and unknown cases were lost during the follow up study. The cause of death of case labeled with an asterisk was unknown. doi:10.1371/journal.pone.0055975.tPCR. For each marker, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated according to previously described formulas [34].All tests were 2 sided, and p-values less than 0.05 were considered statistically significant. Data analysis was performed using Sigma Stat and SPSS ver. 17 software.Mitosis as Source of Bio10236-47-2 markers in Cervical CancerResults Expression Analysis of 8,638 Genes in Cervical CancerThe amount of mRNA transcribed from 8,638 genes was compared between 43 CC samples positive for HPV16 and 12 normal cervical epithelial samples using the HG-Focus microarray. A total of 997 genes were differentially expressed between the cancer and control groups; 600 were upregulated and 397 were downregulated (Table S3). Almost one-half of the upregulated and downregulated genes had FCs in the range of 1.5?.0, and the number of genes in both groups decreased linearly (r = 20.8, p = 0.002) as the FC value increased (Figure 1). The principal component analysis (PCA; data not shown) and the nonsupervised hierarchical clustering (panel A in Figure 2) performed with all 997 gene expression values clearly separated the cancer samples from the control group. However, the expression of many genes was not completely uniform among the cancer samples, especially in the group of upregulated genes (signals shown in red in Figure 2A). Many of those genes were upregulated in some tumors and downregulated in other tumors. This was in contrast to the uniformity of the expression signals in the control group samples. Genes to be tested as markers for screening or as potential therapeutic targets were selected according to D-score rank (a modified t-test, used in SAM), FC or MedChemExpress Methionine enkephalin whether they were previously used as markers for cervical cancer. From the 997 genes associated with the cancer samples, 163 have been previously reported as markers for different types of cancer (IPA, Ingenuity Systems), including MCM2, TOP2A, and CDKN2A, which have been used as markers for diagnosis in cervical cancer [35]. The 997 genes 18325633 were listed in decreasing ordered by D-score (Table S3). A total of 23 genes (18 upregulated and 5 downregulated) were selected for validation by qRT-PCR (marked in bold in Table S3 and Table 2; circles colored in blue and orange in Figure 1). All downregulated genes (CFD, NDN, WISP2, END3, and SLC18A2) and 10 of the 18 upregulated genes (PRC1, CKS2, TYMS, RFC4, RRM2, NUSAP1, MCM2, CCNB2, SMC4, and CDC2) were selected according to Dscore rank. Seven of the remaining upregulated genes are on the list of the 50 best ranked genes, 2 of them are genes that have been previously proposed as markers in CC (CDKN2A and TOP2A), 4 (CDC20, CDKN3, ZWINT, and SYCP2) were selected based on the FC value, and PCNA (Table S3), together with MKI67, which ranked in 139th place, were included because these markers are commonly used to measure cell proliferation. The PCA analysis and hierarchical clustering showed that the 23 selected genes also allowed for segregation of the samples into the 2 different groups. For both the upregulated and downregulated genes, t.Rachy, brachytherapy. Chemo, chemotherapy with Cisplatin. Status alive was registered at the last follow up, death was caused by primary tumor of cervical cancer, except the case labeled with an asterisk, and unknown cases were lost during the follow up study. The cause of death of case labeled with an asterisk was unknown. doi:10.1371/journal.pone.0055975.tPCR. For each marker, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated according to previously described formulas [34].All tests were 2 sided, and p-values less than 0.05 were considered statistically significant. Data analysis was performed using Sigma Stat and SPSS ver. 17 software.Mitosis as Source of Biomarkers in Cervical CancerResults Expression Analysis of 8,638 Genes in Cervical CancerThe amount of mRNA transcribed from 8,638 genes was compared between 43 CC samples positive for HPV16 and 12 normal cervical epithelial samples using the HG-Focus microarray. A total of 997 genes were differentially expressed between the cancer and control groups; 600 were upregulated and 397 were downregulated (Table S3). Almost one-half of the upregulated and downregulated genes had FCs in the range of 1.5?.0, and the number of genes in both groups decreased linearly (r = 20.8, p = 0.002) as the FC value increased (Figure 1). The principal component analysis (PCA; data not shown) and the nonsupervised hierarchical clustering (panel A in Figure 2) performed with all 997 gene expression values clearly separated the cancer samples from the control group. However, the expression of many genes was not completely uniform among the cancer samples, especially in the group of upregulated genes (signals shown in red in Figure 2A). Many of those genes were upregulated in some tumors and downregulated in other tumors. This was in contrast to the uniformity of the expression signals in the control group samples. Genes to be tested as markers for screening or as potential therapeutic targets were selected according to D-score rank (a modified t-test, used in SAM), FC or whether they were previously used as markers for cervical cancer. From the 997 genes associated with the cancer samples, 163 have been previously reported as markers for different types of cancer (IPA, Ingenuity Systems), including MCM2, TOP2A, and CDKN2A, which have been used as markers for diagnosis in cervical cancer [35]. The 997 genes 18325633 were listed in decreasing ordered by D-score (Table S3). A total of 23 genes (18 upregulated and 5 downregulated) were selected for validation by qRT-PCR (marked in bold in Table S3 and Table 2; circles colored in blue and orange in Figure 1). All downregulated genes (CFD, NDN, WISP2, END3, and SLC18A2) and 10 of the 18 upregulated genes (PRC1, CKS2, TYMS, RFC4, RRM2, NUSAP1, MCM2, CCNB2, SMC4, and CDC2) were selected according to Dscore rank. Seven of the remaining upregulated genes are on the list of the 50 best ranked genes, 2 of them are genes that have been previously proposed as markers in CC (CDKN2A and TOP2A), 4 (CDC20, CDKN3, ZWINT, and SYCP2) were selected based on the FC value, and PCNA (Table S3), together with MKI67, which ranked in 139th place, were included because these markers are commonly used to measure cell proliferation. The PCA analysis and hierarchical clustering showed that the 23 selected genes also allowed for segregation of the samples into the 2 different groups. For both the upregulated and downregulated genes, t.

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