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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Cancer Discov. 2013 Aug 1;3(10):10.1158/2159-8290.CD-13-0118. doi: 10.1158/2159-8290.CD-13-0118

Figure 1. HIF1α is an important regulator of metastasis in an autochthonous, genetic model of UPS potentially via PLOD2 modulation.

Figure 1

(A) (Left and Middle Panels) Relative gene expression in human metastatic (N=5) and non-metastatic (N=8) UPS and fibrosarcoma in patients treated as Massachusetts General Hospital (32). HIF1α (P=0.0312) and PLOD2 (P =0.0011) were significantly upregulated in metastastic sarcomas. (Right Panel) qRT-PCR analysis of 10 human UPS patient samples treated at the University of Pennsylvania; P=0.044. (B) Mouse models of sarcoma. LSL-KrasG12D/+;Trp53fl/fl (KP) and LSL-KrasG12D/+;Trp53fl/fl;Hif1αfl/fl (KPH) genotyping showed effective recombination of Hif1αfl alleles in Adeno-Cre initiated tumors. (C) Mice remained tumor free for roughly 40 days, by 90 days all of the mice had developed palpable tumors (volume= 200mm3) KP; n=30, KPH; n=20; (P=0.3755). (D) Primary tumor size. Two weeks after tumors were palpable they had grown 2-8 fold larger, but there was no difference between KP; n=7 and KPH; n =9 tumor growth (P=0.7342) (E) Metastasis free survival in KP; n=33 and KPH; n=28 mice; P=0.0456. Lung metastases were confirmed histologically. (F) Masson's Trichrome and picrosirius red staining of tumor nest areas and blood vessels in primary KP and KPH tumors. Deletion of HIF1α alters collagen in KPH tumors. Masson's Trichrome stains collagen fibers blue. Cells were counterstained in red with Weigert's hematoxylin. (Scale bar;50 μm). (G) Western blot analyses of sarcoma cells derived from KP and KPH tumors. Expression of HIF1α and PLOD2 proteins is hypoxia inducible and abolished when HIF1α is deleted. (H) qRT-PCR analysis of 2 individually derived KP and KPH cell lines. Plod2 mRNA transcription is induced under hypoxic conditions in control cells (KP1; P =0.0284 and KP2; P =0.0391). Deletion of HIF1α abolished hypoxia-induced Plod2 mRNA levels. (I) Western blot of PLOD2 expression in KIA cells and (J) HT-1080 cells. (K) qRT-PCR analyses of KIA cells. Expression of Hif1α and Plod2 is hypoxia inducible (Plod2 qRT-PCR: P =0.0284) and is abolished when HIF1α is deleted (Plod2 qRT-PCR: P =0.0403). (L) HT-1080 cells were evaluated by qRT-PCR as in (K). Expression of Hif1α and Plod2 is hypoxia inducible (Plod2 qRT-PCR: P =0.0006) and is abolished when HIF1α is deleted (Plod2 qRT-PCR: P =0.0210).