HMGA1 overexpression is associated with a particular subset of human breast carcinomas

Breast cancer represents the second leading cause of cancer mortality among American women and accounts for more than 40 000 deaths annually. High-mobility group A1 (HMGA1) expression has been implicated in the pathogenesis and progression of human malignant tumours, including breast carcinomas. The aim of this study was to evaluate HMGA1 detection as an indicator for the diagnosis and prognosis of human breast carcinoma

Metabolic reprogramming identifies the most aggressive lesions at early phases of hepatic carcinogenesis

Metabolic changes are associated with cancer, but whether they are just bystander effects of deregulated oncogenic signaling pathways or characterize early phases of tumorigenesis remains unclear. Here we show in a rat model of hepatocarcinogenesis that early preneoplastic foci and nodules that progress towards hepatocellular carcinoma (HCC) are characterized both by inhibition of oxidative phosphorylation (OXPHOS) and by enhanced glucose utilization to fuel the pentose phosphate pathway (PPP). These changes respectively require increased expression of the mitochondrial chaperone TRAP1 and of the transcription factor NRF2 that induces the expression of the rate-limiting PPP enzyme glucose-6-phosphate dehydrogenase (G6PD), following miR-1 inhibition. Such metabolic rewiring exclusively identifies a subset of aggressive cytokeratin-19 positive preneoplastic hepatocytes and not slowly growing lesions. No such metabolic changes were observed during non-neoplastic liver regeneration occurring after two/third partial hepatectomy. TRAP1 silencing inhibited the colony forming ability of HCC cells while NRF2 silencing decreased G6PD expression and concomitantly increased miR-1; conversely, transfection with miR-1 mimic abolished G6PD expression. Finally, in human HCC patients increased G6PD expression levels correlates with grading, metastasis and poor prognosis. Our results demonstrate that the metabolic deregulation orchestrated by TRAP1 and NRF2 is an early event restricted to the more aggressive preneoplastic lesions

Convergent Evolution of Copy Number Alterations in Multi-Centric Hepatocellular Carcinoma

In the recent years, new molecular methods have been proposed to discriminate multicentric hepatocellular carcinomas (HCC) from intrahepatic metastases. Some of these methods utilize sequencing data to assess similarities between cancer genomes, whilst other achieved the same results with transcriptome and methylome data. Here, we attempt to classify two HCC patients with multi-centric disease using the recall-rates of somatic mutations but find that difficult because their tumors share some chromosome-scale copy-number alterations (CNAs) but little-to-no single-nucleotide variants. To resolve the apparent conundrum, we apply a phasing strategy to test if those shared CNAs are identical by descent. Our findings suggest that the conflicting alterations occur on different homologous chromosomes, which argues for multi-centric origin of respective HCCs

Long non-coding RNAs are key players in Prostate cancer tumorigenesis and drug resistance

"Long non-coding RNAs (lncRNAs) have been characterized as key players in several cancer-associated processes such as tumorigenesis and drug resistance. Emerging evidence indicates that lncRNAs affect initiation and progression of several cancers, including prostate cancer (PCa) and its advanced forms, such as metastatic castration resistant prostate cancer (mCRPC). Among others, H19 is one of the most studied oncogenic lncRNAs in cancer and has been associated with cancer tumorigenesis and progression, via mediating several pathways an acting in different mechanisms of action, including epigenetic and miRNA regulation. We have investigated lncRNA roles in PCa tumorigenesis by analysing the E006AA cell model. Emerging evidence has shown that parental E006AA are not tumorigenic in nude mice, while its derived E006AA-ht, is highly tumorigenic. Via high-throughput RNA sequencing we have shown that E006AA-ht overexpress different lncRNAs compared to E006AA. Interestingly, we found a high upregulation of H19 in E006AA-ht vs the parental cell line. Our in vitro validation has confirmed the sequencing data and further research could unravel a crucial role of H19 in PCa tumorigenesis, opening a new challenging chapter of lncRNAs research. We hypothesize that H19 could be involved in in vivo immunity suppression and our further studies aim at investigating H19 in this molecular and clinical context. Furthermore, we have studied lncRNAs as key players in cancer aggressiveness by using cellular models of mCRPC. From our studies, HORAS5 (i.e. linc00161) emerged as the most consistently upregulated lncRNA in CRPC patient-derived xenografts. This lncRNAs was previously associated with cancer drug-response in osteosarcoma, ovarian cancer and hepatocellular carcinoma. Our study has shown for the first time that HORAS5 promotes drug resistance in CRPC. After a preliminary drug screen, we have selected the chemotherapeutics cabazitaxel for further investigation. Via lentiviral-mediated overexpression and siRNA-based silencing we have regulated HORAS5 expression and analysed cell count and apoptosis of CRPC cells exposed to clinically achievable concentrations of cabazitaxel. The overexpression of HORAS5 increases cabazitaxel resistance, while HORAS5 silencing has an opposite effect, via inhibition of apoptosis. RNA sequencing and RT-qPCR revealed that BCL2A1 is the most upregulated transcript in HORAS5 overexpressing cells exposed to cabazitaxel, and that BCL2A1 silencing decreases cell count and increases caspase activity. Our data suggest that BCL2A1 expression is induced by HORAS5, thereby enhancing CRPC cells resistance to cabazitaxel. Transfection of CRPC cells with HORAS5-targeting ASOs can effectively silence this lncRNA and determine a decrease of cabazitaxel resistance. We have also shown that both HORAS5 and BCL2A1 upregulation results in decreased survival in PCa patients and samples from mCRPC patients treated with taxanes have upregulation of HORAS5. Overall, our studies bring novel insights into crucial roles of lncRNAs in PCa progression and aggressiveness making them emerging targets for cancer treatment at different stages.

VEGFA gene locus analysis across 80 human tumour types reveals gene amplification in several neoplastic entities

Background: Angiogenesis plays a pivotal role in neoplastic growth and metastasis formation. Vascular endothelial growth factor A (VEGFA) is a major player in physiological and tumour-induced angiogenesis and numerous human tumours have been show to overexpress VEGFA. Moreover increased VEGFA gene expression has been found frequently to correlate with tumour progression, recurrences and survival. Interestingly, several studies have demonstrated that gene amplification may result in protein overexpression and that amplification of the therapeutics' target gene can serve as an excellent predictive marker (i.e. HER2 and trastuzumab). However the impact of VEGFA gene amplification has been only recently assessed for some cancer types such as osteosarcoma, colorectal, breast and liver cancer. Aims: This study aimed to assess VEGFA gene amplification status using fluorescent in situ hybridization (FISH) in a large cohort of different tumour entities. Thus, we investigated the incidence of VEGFA amplification using a multi-tumour tissue microarray (TMA) containing 2,837 evaluable specimens from 80 different tumour entities and 31 normal tissue types. Moreover, we validated FISH analysis as reference method to evaluate VEGFA gene status by comparing it to comparative genomic hybridization (CGH). Results: We observed that VEGFA locus amplification and/or polysomy represented a small but regularly detected population in several tumour entities while was not present in normal tissues. VEGFA gene alterations were predominantly observed in hepatocarcinomas, adenocarcinomas of the pancreas and intestine, large cell carcinoma of the lung and in endometrium serous carcinoma. Furthermore our data demonstrated that VEGFA detection by FISH provided highly comparable results to those generated by CGH. Conclusion: Albeit with low percentage, VEGFA amplification is commonly observed across several tumour entities. Furthermore, our results demonstrated that FISH test could be used as a reliable diagnostic tool to evaluate VEGFA gene status in human specimens

Cancer Diagnosis Using a Liquid Biopsy: Challenges and Expectations

The field of cancer diagnostics has recently been impacted by new and exciting developments in the area of liquid biopsy. A liquid biopsy is a minimally invasive alternative to surgical biopsies of solid tissues, typically achieved through the withdrawal of a blood sample or other body fluids, allowing the interrogation of tumor-derived material including circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) fragments that are present at a given time point. In this short review, we discuss a few studies that summarize the state-of-the-art in the liquid biopsy field from a diagnostic perspective, and speculate on current challenges and expectations of implementing liquid biopsy testing for cancer diagnosis and monitoring in the clinical setting

VEGFR-3 is expressed on megakaryocyte precursors in the murine bone marrow and plays a regulatory role in megakaryopoiesis

Introduction VEGFR-3 is a member of the VEGFR receptor tyrosine kinase family. It is expressed on lymphatic endothelial cells (LECs) and plays a central role in the regulation of lymphangiogenesis. 1 On binding to its ligands, VEGF-C and VEGF-D, VEGFR-3 is activated and orchestrates the outgrowth of lymphatic vessels. During murine hematopoiesis, Sca-1 ϩ hematopoietic stem cells give rise to the precursors of all hematopoietic lineages. 10 Megakaryocytes develop from CD34 ϩ progenitors. Methods Cell culture HEL cells were obtained from DSMZ and cultivated in RPMI (Gibco-BRL) containing 10% FCS and 1% penicillin-streptomycin. Differentiation was induced with 10nM tetradecanoyl phorbol acetate (TPA; Sigma-Aldrich). Primary human microvascular LECs (Cambrex) from the dermis (HMVECdLyNeo) were cultivated in EGM-2MV (Lonza) and 5% FCS supplemented with growth factors provided by the manufacturer. Bovine lymphatic endothelial cells were cultivated in DMEM (Gibco-BRL) containing 20% FCS and 1% penicillin-streptomycin on gelatin-coated plastic. HEK-293 cells were cultivated in DMEM supplemented with 10% FCS and 1% penicillin-streptomycin. Western blot analysis Cell lysates were analyzed using standard Western blotting techniques. The membranes were probed with Abs specific for VEGFR-3 (R&D Systems), CD31 (Santa Cruz Biotechnology), CD34 (Abcam), CD42a (Santa Cruz Biotechnology), CD61 (R&D Systems), CD144 (Santa Cruz Biotechnology), or GpA (International Blood Group Reference Laboratory). Probing with hypoxanthine phosphoribosyltransferase (HPRT) Abs (Santa Cruz Biotechnology) served as a loading control. PCR analysis RNA was prepared using peqGOLD RNAPure (PeqLab). Synthesis of cDNA using Superscript II (Invitrogen) was performed according to the manufacturer's recommendations. For PCR, cDNAs were amplified as follows: 94°C for 30 seconds, 60°C for 30 seconds, and 72°C for 90 seconds (VEGFR-2, Prox1, LYVE-1, Podoplanin, HPRT, Fli-1, Fog-2, Gata-2, and Elf-1) or 94°C for 30 seconds, 54°C for 30 seconds, and 72°C for 90 seconds (VEGFR-3). Details of the primers used are in supplemental Methods (available on the Blood Web site; see the Supplemental Materials link at the top of the online article). Tubule formation on collagen gels Collagen type 1 was prepared from rat tails. Tendons were isolated, dissolved in acetic acid, then filtered, lyophilized, and redissolved in 0.1% acetic acid at 4 mg/mL. Cells were seeded on collagen gels (2 mg/mL) and cultured in the presence of 30 ng/mL of VEGF 165 (Promokine) for 8 days. Tubule formation was analyzed as described previously. 22 Immunohistochemistry For the immunohistochemical analysis of VEGFR-3 expression in the BM, cryosections of decalcified murine femurs embedded in tissue-freezing medium (Leica) were fixed in acetone and stained with VEGFR-3 Abs (eBiosciences). The stained sections were then analysed at room temperature using an Axioskop (Zeiss) equipped with a PlanNeoflur 20ϫ/0.50 and an Axiocam (Zeiss) and Axiovision software (Ziess). MACS BM cells isolated from femurs and tibias of C57BL/6 mice were treated with Fc-block (BD Biosciences) and then incubated with Abs against VEGFR-3 (R&D Systems), Sca-1, CD41, or CD38 (BD Biosciences), followed by specific secondary MACS Abs (Miltenyi-Biotec) according to the manufacturerЈs recommendations. Cell populations were then either enriched or depleted for the labeled epitope using LS or LD columns (Miltenyi-Biotec), respectively. The purity of the sorted populations was controlled by flow cytometry. CD42 FACS BM was isolated from femurs and tibias of C57BL/6 mice and stained with Abs specific for VEGFR-3 (R&D Systems) and/or CD42a (Emfret) and analyzed by FACS. Lethal irradiation and BM transplantation C57BL/6 mice were irradiated with lethal doses (9 Gy) from a ␥ source. After 24 hours, the mice were all transplanted in parallel by IV injection with either complete BM, BM depleted of VEGFR-3 ϩ cells, or BM mock depleted with an appropriate isotype control using MACS. EDTA blood samples were taken from all animals on days 0, Isolation and culture of primary murine BM cells BM was isolated from femurs and tibias of C57BL6 mice. After lysis of RBCs with ammonium-chloride-potassium buffer, the cells were transferred to IMDM (Gibco-BRL) supplemented with 1% penicillin/streptomycin, 10% HEK-293 cell-conditioned DMEM, Nutridoma SP (Roche), L-glutamine, and 100 pg/mL of recombinant murine TPO (RDI Diagnostics). Depending on the experiment, the cells were cultured with either 100 g/mL of mF4-31C1 VEGFR-3-blocking Abs (kindly provided by ImClone Systems), 100 g/mL of rat IgG isotype control, or 400 ng/mL of VEGF-C-Cys, a mutant form of VEGF-C that activates VEGFR-3 but not VEGFR-2. Long-term injections C57BL/6 mice were injected daily with 25 g of VEGF-C-Cys for 3 weeks. Blood was taken on days 0, 3, 7, 10, 14, 17, and 21. In the blocking Ab experiments, mice were injected with 600 g/animal/injection of mF4-31C1 VEGFR-3-blocking Ab, isotype control Ig, or PBS on a MondayWednesday-Friday schedule for 6 weeks. Blood was taken on days 0, Recovery kinetics after sublethal irradiation Experimental C57BL/6 mice were sublethally irradiated (4.5 Gy) in a ␥ source. They were then either injected daily with VEGF-C-Cys (25 g/animal/injection) or PBS or were intraperitoneally injected with 600 g/animal/injection of mF4-31C1 VEGFR-3-blocking Abs, isotype control Ig, or PBS every other day. Blood was taken on days 0, 7, 11, 14, 18, and 21 after irradiation and analyzed. In each experiment, all animals were treated at the same time and on the same day and all animals were bled at each time point. BM was isolated from femurs and tibias 20 days after irradiation, and the number and ploidy of CD41 ϩ cells in the BM was assessed. Significance was tested using 2-tailed unpaired t tests assuming equal variance. TPO administration C57BL/6 mice were administered with 5 g of recombinant murine TPO (RDI), followed by daily injections of either 25 g of VEGF-C-Cys or PBS. One group received only PBS throughout. Blood was taken and analyzed 0, 3, 5, 7, and 10 days after TPO administration. All animals were treated at the same time and on the same day and all animals were bled at each time point. After 10 days, the animals were killed and the number and ploidy of CD41 ϩ 1900 THIELE et al BLOOD, 30 AUGUST 2012 ⅐ VOLUME 120, NUMBER 9 For personal use only. on October 6, 2016. by guest www.bloodjournal.org From cells in the BM was assessed. Significance was tested using 2-tailed unpaired t tests assuming equal variance. 5-FU treatment C57BL/6 mice were intraperitoneally injected with a single dose of 5-FU (Sigma-Aldrich) at 150 mg/kg. Control mice remained untreated. The 5-FU-treated mice then received daily injections of either 25 g of VEGF-C-Cys or PBS throughout the experiment. Blood was taken and analyzed 0, All animal experiments were approved by the local regulatory authorities and were performed according to German legal requirements. Results Expression of VEGFR-3 and other lymphatic endothelial markers is up-regulated on phorbol diester-induced megakaryocytic differentiation of HEL cells VEGFR-3 is widely used as a marker for lymphatic endothelium. Originally, however, the receptor was cloned from the HEL cell line. 7 This cell line can be induced to differentiate into the erythrocyte lineage by EPO treatment 23 and into the megakaryocyte lineage in response to TPA. Consistent with the notion that HEL cells differentiate into the megakaryocyte lineage on TPA treatment, we detected strong up-regulation of several markers and transcription factors associated with megakaryocytic differentiation A survey of the literature revealed that virtually all markers described to date as being expressed on megakaryocytes can also be expressed on endothelial cells (supplemental These observations raised the question of whether HEL cells really undergo megakaryocytic differentiation after TPA treatment or if they adopt an endothelial phenotype with LEC characteristics. To address this point, we investigated whether TPA-treated HEL cells are capable of forming capillaries, reasoning that if the cells differentiated into endothelial cells, this should be the case. However, in contrast to control bovine LECs, TPA-treated HEL cells could not be induced to form capillaries VEGFR-3 IN MEGAKARYOPOIESIS 1901 BLOOD, 30 AUGUST 2012 ⅐ VOLUME 120, NUMBER 9 For personal use only. on October 6, 2016. by guest www.bloodjournal.org From VEGFR-3 is expressed on megakaryocytic progenitors through to the promegakaryoblast stage in the BM The up-regulation of VEGFR-3 during HEL cell megakaryocytic differentiation suggested to us that VEGFR-3 may play a role in megakaryopoiesis. Because of the limited megakaryocytic differentiation capacity of HEL cells and their cancerous nature, we explored this possibility further using murine BM. First we characterized VEGFR-3 expression in the BM. FACS staining revealed that approximately 2% of murine BM cells were VEGFR-3 ϩ ( To define further the stages of megakaryopoiesis during which VEGFR-3 is expressed, costainings with the stem cell marker Sca-1 and with CD38, CD41, and VEGFR-3 were performed. Expression of Sca-1 is lost during myeloid differentiation. 25 CD38 expression, in turn, is increased early in megakaryopoiesis from the BFU-MK stage on. These observations suggested to us that VEGFR-3 might be expressed on hematopoietic stem cells through to the promegakaryoblast stage. However, Sca1 is not just expressed on hematopoietic stem cells, but also on the immediate progenitors arising from the stem cells. These data are consistent with the notion that VEGFR-3 is not expressed on hematopoietic stem cells, but rather on megakaryocyte precursors through to the premegakaryoblast stage, and that VEGFR-3 expression is lost as megakaryocytes further mature. This notion is further substantiated by the observation that VEGFR-3 ϩ BM cells coexpressed CD42, a marker for megakaryocytes that is not expressed on hematopoietic precursor cells (supplemental Manipulation of VEGFR-3 influences megakaryopoiesis in vitro To examine the role that VEGFR-3 plays during megakaryopoiesis, we cultivated primary murine BM cells with physiologic concentrations of TPO to maintain the megakaryocyte precursors. The cells were grown for 3 days in the presence or absence of VEGF-C-Cys, a mutant form of VEGF-C that specifically activates VEGFR-3 but not VEGFR-2, 20 because VEGFR-2 is also present on megakaryocytic cells. Our data suggest that the specific activation of VEGFR-3 during megakaryopoiesis impairs the transition to polyploid stages, whereas blocking the receptor promotes differentiation and endoreplication. For personal use only. on October 6, 2016. by guest www.bloodjournal.org From Neither activation nor blocking of VEGFR-3 influences steady-state megakaryopoiesis or thrombopoiesis in vivo To study the potential effects of VEGFR-3 manipulation on megakaryopoiesis and thrombopoiesis in vivo, we first injected VEGF-C-Cys to activate VEGFR-3, or PBS as a control, into mice on a daily basis for 3 weeks. Thrombocyte concentrations in the blood were monitored regularly. After 3 weeks of treatment, the mice were killed. BM cells were isolated and stained for CD41 and DNA content to evaluate the number and ploidy of the CD41 ϩ population. We observed a significant decrease in apoptotic CD41 ϩ BM cells in the VEGF-C-Cys-treated group (P Ͻ .01), a trend toward reduced polyploidy, and an increase in 2n CD41 ϩ cells, which were consistent with our in vitro observations. VEGF-C-Cys had no effect on platelet counts or the number of CD41 ϩ cells in the BM (supplemental To determine the effect of inhibiting VEGFR-3 activation on megakaryopoiesis and thrombopoiesis in vivo, mice were injected daily with VEGFR-3-blocking Abs or an appropriate isotype control for 6 weeks. Platelet counts were monitored regularly and the numbers and ploidy distribution of CD41 ϩ BM cells were analyzed at the end of the experiment. Under these conditions, no effects on the measured parameters were observed (supplemental Activation of VEGFR-3 increases platelet counts in TPO-stimulated animals, modulates 5-FU-induced thrombocytopenia and thrombocytosis, and influences ploidy distribution and numbers of CD41 ؉ BM cells after sublethal irradiation Thrombocyte homeostasis is tightly controlled in mammals, and alternative mechanisms exist that can compensate for perturbation . FACS analysis showed that 1.85% Ϯ 0.31% SEM (n ϭ 9) of the murine BM cells expressed VEGFR-3. Dot plots of 1 representative experiment are depicted. Density plots were used to define a region in which 95% (the 2 outer contours) of the negative control events were excluded. The region was then applied to a plot displaying the stained sample. The number of positive events in both the negative control and the actual sample was then assessed. The percentage of true positive cells was calculated by subtraction of the number of events in the negative control within the defined region from the number of events found in the same region for the actual sample. Identical numbers of events were acquired. (B) VEGFR-3 is expressed on isolated mononuclear cells in the murine BM. Sections of murine femurs were stained with VEGFR-3-specific Abs (left panel, VEGFR-3; right panel, control). MK indicates megakaryocyte. Scale bars indicate 100 m. (C) Ploidy of VEGFR-3 ϩ cells in the murine BM. VEGFR-3 ϩ BM cells were enriched by MACS and then analyzed in FACS. As a control, cells were treated with an appropriate isotype control. Clumping cells mimicking polyploidy were excluded from the analysis by appropriate gating strategies. The resulting histogram plot shows the DNA content of VEGFR-3 ϩ cells. Dot plots of the DNA content of the cells were used for the quantification of VEGFR-3 ϩ and isotype-treated cells within different ploidy classes or cell cycle stages, respectively (a detailed scheme of the gating strategy is provided in supplementa

Syrosingopine sensitizes cancer cells to killing by metformin

We report that the anticancer activity of the widely used diabetic drug metformin is strongly potentiated by syrosingopine. Synthetic lethality elicited by combining the two drugs is synergistic and specific to transformed cells. This effect is unrelated to syrosingopine's known role as an inhibitor of the vesicular monoamine transporters. Syrosingopine binds to the glycolytic enzyme α-enolase in vitro, and the expression of the γ-enolase isoform correlates with nonresponsiveness to the drug combination. Syrosingopine sensitized cancer cells to metformin and its more potent derivative phenformin far below the individual toxic threshold of each compound. Thus, combining syrosingopine and codrugs is a promising therapeutic strategy for clinical application for the treatment of cancer

LncRNA HORAS5 promotes taxane resistance in castration-resistant prostate cancer via a BCL2A1-dependent mechanism.

Background: Castration-resistant prostate cancer (CRPC) is an incurable malignancy. Long noncoding RNAs (lncRNAs) play key roles in drug resistance. Materials & methods: LncRNA HORAS5 role in cabazitaxel resistance (i.e., cell-count, IC50 and caspase activity) was studied via lentiviral-mediated overexpression and siRNA-based knockdown. Genes expression was analyzed with RNA-sequencing, reverse transcription quantitative PCR (RT-qPCR) and western blot. HORAS5 expression was queried in clinical database. Results: Cabazitaxel increased HORAS5 expression that upregulated BCL2A1, thereby protecting CRPC cells from cabazitaxel-induced apoptosis. BCL2A1 knockdown decreased cell-count and increased apoptosis in CRPC cells. HORAS5-targeting antisense oligonucleotide decreased cabazitaxel IC50. In CRPC clinical samples, HORAS5 expression increased upon taxane treatment. Conclusion:HORAS5 stimulates the expression of BCL2A1 thereby decreasing apoptosis and enhancing cabazitaxel resistance in CRPC cells

High expression of HOXA13 correlates with poorly differentiated hepatocellular carcinomas and modulates sorafenib response in in vitro models

Hepatocellular carcinoma (HCC) represents the fifth and ninth cause of mortality among male and female cancer patients, respectively and typically arises on a background of a cirrhotic liver. HCC develops in a multi-step process, often encompassing chronic liver injury, steatosis and cirrhosis eventually leading to the malignant transformation of hepatocytes. Aberrant expression of the class I homeobox gene family (HOX), a group of genes crucial in embryogenesis, has been reported in a variety of malignancies including solid tumors. Among HOX genes, HOXA13 is most overexpressed in HCC and is known to be directly regulated by the long non-coding RNA HOTTIP. In this study, taking advantage of a tissue microarray containing 305 tissue specimens, we found that HOXA13 protein expression increased monotonically from normal liver to cirrhotic liver to HCC and that HOXA13-positive HCCs were preferentially poorly differentiated and had fewer E-cadherin-positive cells. In two independent cohorts, patients with HOXA13-positive HCC had worse overall survival than those with HOXA13-negative HCC. Using HOXA13 immunohistochemistry and HOTTIP RNA in situ hybridization on consecutive sections of 16 resected HCCs, we demonstrated that HOXA13 and HOTTIP were expressed in the same neoplastic hepatocyte populations. Stable overexpression of HOXA13 in liver cancer cell lines resulted in increased colony formation on soft agar and migration potential as well as reduced sensitivity to sorafenib in vitro. Our results provide compelling evidence of a role for HOXA13 in HCC development and highlight for the first time its ability to modulate response to sorafenib