Dian Cq 8.61 8.29 13.03 8.87 13.75 15.7 13.21 12.53 8.43 7.54 12.65 10…

Dian Cq 8.61 8.29 13.03 8.87 13.75 15.7 13.21 12.53 8.43 7.54 12.65 10.48 6.62 9.34 12.75 8.49 12.29 11.61 12.33 5.52 16.92 13.45 10.9 10.97 8.27 7.26 10.2 12.4 9.8 11.77 11.93 9.23 8.77 9.61 1.85 0.38 0.18 0.02 6.3 0.23 6.2 0.02 0.4 0.5 1.19 0.81 0.1 0.004 0.01 <0.001 0.81 0.66 0.04 <0.001 7.9 0.04 0.2 0.02 0.5 0.47 0.64 0.43 4.37 0.03 0.07 0.01 Median fold change 0.75 p value 0.91 Mutated CLL Vs. unmutated CLL IGHV Mutation status Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Mutated Unmutated Median Cq 9.8 7.46 12.85 12.63 13.51 13.71 12.93 13.34 8.86 8.37 12.65 12.65 6.5 5.98 12.18 12.7 12.8 9.28 12.32 11.72 16.42 16.91 10.57 11.18 8.4 8.08 9.78 10.26 9.2 8.91 11.65 11.92 8.59 8.72 Median fold change 0.31 1.73 0.05 0.06 3.89 3.2 0.62 0.59 0.39 0.57 0.25 0.26 6.96 11.4 0.05 0.03 0.37 4.61 0.01 0.01 0.08 0.08 1.15 0.82 0.44 0.53 6.59 4.6 4.79 6.66 0.18 0.16 1.85 1.75 0.78 0.99 0.56 0.42 0.54 0.56 0.67 0.47 <0.001 0.28 0.32 0.7 0.37 0.6 0.45 0.85 p value <0.The statistically significant p values are shown in italics[11]], or in other malignancies [BIK [34], SPRY2 [35], TBX2 [36, 37], TSHZ3 [38, 39]]. As expected, MEIS1, PMEPA1, SOX7, SPRY1, CDK6, TBX2 were significantly downregulated (p < 0.05) while SPRY2 (p = 0.016), VIPR1 (p = 0.04) and ID4 (p = 0.03) were significantly upregulated in CLL cells as compared to healthy B-cells. Though not significant, AXIN2 was upregulated and TNRC18,NFATC1 and BIK were downregulated in CLL as compared to healthy CD19+ cells (Table 2). The expression of only CRY1 and PAX9 differed significantly (p < 0.05) with respect to the IGHV mutation status (Table 2, Fig. 4) The status of hypomethylation of PAX9 among unmutated CLL was confirmed through bisulfite genome sequencing of CpG island 3 in close proximity to PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/18111632 CpGRani et al. Clinical Epigenetics (2017) 9:Page 8 ofFig. 4 Box-plot representation of mRNA expression fold change as assessed by RQ-PCR for a CRY1 gene in CLL and its IGHV mutated and unmutated subgroups b PAX9 gene in CLL and its IGHV mutated and unmutated subgroups. Box-plot graphs show median (middle line), interquartile range (box), 25?5th percentile (whiskers) and PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9221828 statistically significant difference (p value) estimated in comparison between different groups(Fig. 1).While CpG island 7 did not reveal any significant difference in methylation levels, the average methylation at CpG island 3 was found to be 52.74 in mutated while 24.72 among unmutated CLL. This further corroborates with the reduced expression of PAX9 in unmutated group of CLL patients as established through microarray-based observations.Association between gene expression and clinical outcomeOf the 17 genes evaluated for mRNA expression, CRY1 (p = 0.008) and PAX9 (p < 0.001) were expressed at higher levels in Rai stage I and II as compared to stage 0. A progressive increase in the expression of CRY1 (p = 0.004) and PAX9 (p < 0.001) was observed in increasing IPI score categories ranging from 1 to 4. We further explored the association between expression level of candidate genes with relative risk of treatment initiation, TTFT and OS (Table 3). The relative risk of treatment initiation was significantly higher with high expression of PAX9 (p = 0.001) or CRY1 (p = 0.005). The high expressio.