Fig. 7: Novel tWT1 transcripts are expressed in human cancers and are indicative of poor long-term survival in ovarian cancers. | Cancer Gene Therapy

Fig. 7: Novel tWT1 transcripts are expressed in human cancers and are indicative of poor long-term survival in ovarian cancers.

From: Long-term severe hypoxia adaptation induces non-canonical EMT and a novel Wilms Tumor 1 (WT1) isoform

Fig. 7

A tWT1 expression in human melanoma cell lines undergoing LTHY as measured by qPCR. Expression presented as relative to fold change (FC) to the 5% O2 condition B tWT1 expression in ZR75 cells during LTHY incubation in the absence of estradiol (E2), relative to the 5% O2 timepoint. C Left: sunburst plot of Leucegene samples based on WT1 gene and isoform expression. Iso WT1 denotes the number of samples where WT1 isoform calling could be performed. Right: Breakdown of tWT1-G/P expression levels in tWT1 expressing Leucegene samples. Isoform calling was done using km and the isoform specific difference between G and P, using km’s Expectation-Maximization algorithm. D tWT1-GFP expression levels as determined by FACS in HEK293T cells. tWT1-GFP expression was induced by 2 µg/mL DOX for 36 hs at the 0.5% O2 timepoint of the LTHY protocol. E Left: Sunburst plot of TCGA-OV samples by WT1 isoform expression. Right: Breakdown of tWT1-G/P expression levels in tWT1-G/P expressing TCGA-OV samples. Isoform calling was done using km and the isoform-specific difference between G and P, using km’s Expectation-Maximization algorithm. F WT1 and tWT1 mRNA expression in human ovarian cancer cell line OVCAR3 after completing the LTHY adaptation, relative to normoxia. G RT-PCR of human tWT1 isoforms from normoxic OVCAR3 cDNA. H Delta-CT values calculated from OVCAR3 and TOV3291G cells in normoxic conditions using RPL10 as housekeeping gene to assess baseline expression. I WT1 and tWT1 mRNA expression in human ovarian cancer cell line TOV3291G after completing LTHY, relative to normoxia. Data presented as relative expression to normoxic condition, J Survival curve analyses for TCGA-OV (ovarian cancer) based on WT1 expression subsets. Left: Kaplan-Meier estimation survival curve of TCGA-OV samples, comparing samples which express any isoform of WT1 versus those with no WT1 expression. Curves are not significantly different (p = 0.26). Right: Kaplan-Meier estimation survival curve of TCGA-OV samples, comparing samples which express tWT1 isoforms (isoforms G or P) versus those which exclusively express canonical WT1 isoforms. Two-sided p-value of the whole curve is 0.12. Two-sided p-value of post-median data (126 observations) is 0.039. WT1 expression and isoform calling were determined by detection of exons 1, 1a, 2, 4, 7, and isoform G exon 1 by km.

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