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Collective migration is important to embryonic development and cancer metastasis, but migratory and nonmigratory cell fate discrimination by differential activity of signal pathways remains elusive. In

Hodgkin lymphoma (HL) is composed of neoplastic Hodgkin and Reed‐Sternberg cells in an inflammatory background. The neoplastic cells are derived from germinal center B cells that, in most cases, are infected by Epstein‐Barr virus (EBV), which may play a role in tumorigenesis. Given that EBV‐latent membrane protein 1 (LMP1) regulates autophagy in B cells, we explored the role of autophagy mediated by EBV or LMP1 in HL. We found that EBV‐LMP1 transfection in HL cells induced a modest increase in autophagy signals, attenuated starvation‐induced autophagic stress, and alleviated autophagy inhibition‐ or doxorubicin‐induced cell death. LMP1 knockdown leads to decreased autophagy LC3 signals. A xenograft mouse model further showed that EBV infection significantly increased expression of the autophagy marker LC3 in HL cells. Clinically, LC3 was expressed in 15% (19/127) of HL samples, but was absent in all cases of nodular lymphocyte‐predominant and lymphocyte‐rich classic HL cases. Although expression of LC3 was not correlated with EBV status or clinical outcome, autophagic blockade effectively eradicated LMP1‐positive HL xenografts with better efficacy than LMP1‐negative HL xenografts. Collectively, these results suggest that EBV‐LMP1 enhances autophagy and promotes the viability of HL cells. Autophagic inhibition may be a potential therapeutic strategy for treating patients with HL, especially EBV‐positive cases., EBV‐LMP1 enhances autophagy and promotes viability of Hodgkin lymphoma cells. Autophagic inhibition may be a potential therapeutic strategy for treating patients with Hodgkin lymphoma, especially EBV‐positive cases.

Diffuse large B-cell lymphoma (DLBCL) may present initially in bone marrow, liver and spleen without any lymphadenopathy (referred to as BLS-type DLBCL), which is aggressive and frequently associated with hemophagocytic syndrome. Its tumorigenesis and molecular mechanisms warrant clarification. By gene microarray profiling with bioinformatics analysis, we found higher expression of the stem cell markers HOXA9 and NANOG, as well as BMP8B, CCR6 and S100A8 in BLS-type than conventional DLBCL. We further validated expression of these markers in a large cohort of DLBCL including BLS-type cases and found that expression of HOXA9 and NANOG correlated with inferior outcome and poor prognostic parameters. Functional studies with gene-overexpressed and gene-silenced DLBCL cell lines showed that expression of NANOG and HOXA9 promoted cell viability and inhibited apoptosis through suppression of G2 arrest in vitro and enhanced tumor formation and hepatosplenic infiltration in a tail-vein-injected mouse model. Additionally, HOXA9-transfected tumor cells showed significantly increased soft-agar clonogenic ability and tumor sphere formation. Interestingly, B cells with higher CCR6 expression revealed a higher chemotactic migration for CCL20. Taken together, our findings support the concept that tumor or precursor cells of BLS-type DLBCL are attracted by chemotaxis and home to the bone marrow, where the microenvironment promotes the expression of stem cell characteristics and aggressiveness of tumor cells.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma worldwide, accounting for about 40% of cases. The role of cytokines in the pathogenesis of lymphomas has been rarely addressed, although cytokines have a close immunological relationship with lymphocytes. We observed overexpression of interleukin (IL)-20 in reactive germinal centres (GCs) leading us to hypothesise that IL-20 may play a role in lymphomagenesis. In this study, we surveyed for IL-20 expression in various types of lymphoma and found that IL-20 was expressed most frequently in follicular lymphoma (94%), but also in Burkitt lymphoma (81%), mantle cell lymphoma (57%), nodal marginal zone lymphoma (56%), Hodgkin lymphomas (50%), small lymphocytic lymphoma (50%) and diffuse large B-cell lymphoma (DLBCL, 48%). IL-20 was not expressed in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), lymphoplasmacytic lymphoma, and plasmacytoma. T-cell lymphomas were largely negative for IL-20 expression, except for anaplastic large cell lymphoma (ALCL, 61%), which frequently expressed IL-20, especially in cutaneous ALCL, and showed an inverse association with ALK expression (p=0.024). We further tested IL-20 expression in another large cohort of DLBCL and found IL-20 expression more frequently in germinal centre B-cell (GCB) than in non-GCB subtype [16/26 (62%) versus 24/64 (38%), p=0.038]. In this cohort, IL-20 was associated with a lower rate of extranodal involvement (p=0.009), bone marrow involvement (p=0.040), and better overall survival (p=0.020). Mechanistically, IL-20 overexpression promoted G1 cell cycle arrest and subsequent apoptosis of DLBCL cells and vice versa in vitro. We conclude that IL-20 may be involved in lymphomagenesis and may be useful as a prognostic marker in patients with DLBCL. In addition, IL-20 plays an inhibitory role in DLBCL growth, probably through cell cycle regulation.

Introduction: Chemoresistance is one of the major challenges for cancer therapy. Oxaliplatin is the first-line therapy for most cancers, including colorectal cancer (CRC). However, the underlying mechanisms of oxaliplatin resistance remain elusive. Previous reports indicated that microRNAs might regulate the development of CRC. Dicer, an endonuclease involved in miRNA biogenesis, may play a critical role in oxaliplatin resistance. Here, we investigated the mechanism of Dicer-mediated oxaliplatin resistance in CRC. Materials and Methods: Oxaliplatin-resistant CRC cells were established. Colony formation, migration, invasion, and sphere formation assays were performed to characterize the properties of oxaliplatin-resistant CRC cells. cDNA microarray and NGS were used to identify the altered expression of mRNAs and miRNAs in oxaliplatin-resistant cells. RT-qPCR determined gene expression. A connectivity map (CMAP) database was used to find the potential drugs which can reduce cell viability in oxaliplatin-resistant cells. CCK-8 assay was used to measure the anti-tumor effect of those potential drugs in oxaliplatin-resistant CRC cells. PANTHER was used to find the potential pathways that may involve drug resistance in CRC. Both reporter assay and western blot analyzed the activation of the Wnt/β-catenin and MAPK signaling pathways. Results: Dicer is overexpressed in oxaliplatin-resistant CRC cells and CRC recurrent patients. Overexpressed Dicer attenuated the cytotoxicity of oxaliplatin and promoted the cell migration, invasion, and stem cell properties of the oxaliplatin-resistant cells. Knocked-down of Dicer restored the chemosensitivity and decreased all of those abilities in oxaliplatin-resistant CRC cells. The expression of several potential mRNAs and microRNAs was changed in drug-resistant cells and chemo-resistant CRC patients' plasma. MAPK and Wnt/β-catenin pathways are hyper-activated in oxaliplatin-resistant CRC cells. PRI-724, a Wnt/β-catenin pathway inhibitor, can increase the cell apoptosis of oxaliplatin-resistant CRC cells. Trametinib, the MEK inhibitor, can induce cell apoptosis in oxaliplatin-resistant CRC cells and present an anti-tumor effect in oxaliplatin-resistant CRC cell-derived xenograft tumors. Knocked-down expression of Dicer did not affect the Wnt/β-catenin pathway but attenuated the ERK activation. A combination of oxaliplatin and trametinib can synergistically inhibit tumor growth and reverses oxaliplatin resistance in vitro and in vivo. Conclusion: Dicer might be an acquired factor contributing to cancer malignancies in oxaliplatin-resistant CRC cells via activating the ERK pathway. The activation of the MAPK signaling pathway is Dicer-dependent in oxaliplatin-resistant CRC cells. Wnt/β-catenin inhibitors and ERK inhibitors have an anti-tumor effect in oxaliplatin-resistant cells. Citation Format: Yu-Heng Kuan, Ming-Ern Wong, Liang-Yi Hung. Dicer-dependent of ERK activation promotes the acquired oxaliplatin resistance in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 420.

Introduction: The tumor microenvironment (TME) is composed of different cell types and is involved in tumor progression. Several immunosuppressive signaling pathways can promote tumor growth by inhibiting anti-tumor immunity in TEM. Therefore, targeting the immunosuppressive signaling pathways is a potential cancer treatment. Recent studies reported that inhibition of oncoproteins could lead to reduced anti-tumor immunity in several cancers; therefore, it’s critical to identify the potential oncoproteins which can lead to enhanced anti-tumor immunity. Aurora-A is a serine/threonine kinase that plays an important role in the centrosome, cell mitosis, and chromosome stability. Previous studies indicated that cancer patients with poor prognoses are associated with a high Aurora-A level in tumor cells. Inhibition of Aurora-A is considered as a promising strategy for cancer treatment. However, Aurora-A inhibitors have not had a breakthrough in phase II or phase III clinical trials. The role of Aurora-A in tumor immunity is still unclear. Here, we investigated the role of Aurora-A in tumor immunity. Materials and Methods: shRNA was used to knock down Aurora-A, and Alisertib was added to inhibit Aurora-A. RT-qPCR and western blot were performed to determine the IL-16 expression. The secretion of cytokines was measured by cytokine array and ELISA. BALB/c and NOD/SCID mice were subcutaneously injected with CT26 and then administered daily with Alisertib or treated with anti-IL-16 antibody. The tumor-infiltrating immune populations were analyzed by flow cytometry. IHC and H/E stain were performed to analyze Aurora-A expression and lymphocyte infiltration in human CRC specimens. Results: CRC patients with lower lymphocyte infiltration and higher Aurora-A are associated with poor prognosis and vice versa. Knockdown of Aurora-A in CT26 promotes tumor growth in immunocompetent mice by inhibiting the infiltration and cytotoxic activity of CD8+ T cells. Aurora-A negatively regulates IL-16 expression in a kinase-dependent manner in CRC cells. Blockade of IL-16 by anti-IL-16 antibody can reverse the tumor growth and CD8+ T cell cytotoxic activity in Aurora-A knocked-down CT26 tumors. Alisertib can inhibit CT26 tumor growth and promote IL-16 secretion in TME in mice. A combination of Alisertib and IL-16 antibody can promote the therapeutic effect of Alisertib in CT26 tumors. Conclusion: Tumor-intrinsic Aurora-A plays a diverse role in tumor progression depending on the status of lymphocyte infiltration in TME. Aurora-A can promote anti-tumor immunity via negatively regulating IL-16 in a kinase activity-dependent manner in hot CRC tumors. In tumors with high lymphocyte infiltration, Aurora-A inhibition can suppress cancer cell growth and reduce anti-tumor immunity; the combination of Aurora-A inhibitors and IL-16 antibodies may provide a novel and effective strategy for cancer therapy in hot tumors. Citation Format: Shiang-Jie Yang, Ming-Derg Lai, Liang-Yi Hung. Aurora-A kinase promotes anti-tumor immunity in high lymphocyte-infiltrated colorectal cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5169.

Introduction: Dysregulation of cell cycle plays a crucial role during hepatocarcinogenesis. RNA polymerase III is hyperactive and the products of RNA polymerase III are overexpressed in different types of tumors, including of hepatocellular carcinoma (HCC). Maf1 is an RNA polymerase III repressor that can reduce tRNA biosynthesis via inhibiting the interaction between RNA polymerase and TFIIIB and TFIIIC in nucleus. Maf1 can also inhibit tumor growth and AKT/mTOR signaling pathway via enhancing PTEN expression mTOR can phosphorylate Maf1 at Ser-75, and the phosphorylated Maf1 will be exported into cytosol and then ubiquitinated; therefore, promote protein synthesis and cell cycle progression in HCC. Aurora-A is a serine/threonine kinase and plays a vital role in regulating cell cycle progression. Overexpression of Aurora-A has been observed in HCC. However, the relationship between Maf1 and Aurora-A remains unclear. Materials and Methods: HCC cells were transfected with pGFP-Aurora-A and pHA-Maf1 to investigate their correlation. Alisertib was added to inhibit Aurora-A activity. RT-qPCR was performed to determine the RNA expression of Aurora-A, Maf1, pre-tRNALeu and pre-tRNAiMet. Western blot was performed to analyze expression levels of Maf1 and Aurora-A. Cycloheximide was added to observe the protein stability of Maf1. The protein interaction between Aurora-A and Maf1 was determined by co-immunoprecipitation and proximity ligation assay. The subcellular localization of Maf1 was observed by cell fraction assay and immunofluorescence. The cell proliferation was analyzed by CCK-8. Results: Aurora-A can interact with C domain of Maf1, and promote the cytosolic localization and protein stability of Maf1 in a kinase-dependent manner. Two putative Aurora-A phosphorylation sites of Maf1 were mutated, and we found that one of them can result in a nucleolar localization and a reduced protein stability in Maf1. Surprisingly, we also found that overexpression of the Maf1 mutant (Maf1-MT) cannot suppress tRNA biosynthesis compared to WT Maf1. Maf1-overexpressed HCC cells were more sensitive to Aurora-A inhibitors treatment. Conclusion: Aurora-A can promote the protein stability and cytosolic localization of Maf1 via interacting with its C domain in a kinase-dependent manner in HCC cells. Aurora-A phosphorylation site mutant of Maf1 can promote nucleolar localization but lost the RNA polymerase III suppressor function. In Maf1 overexpressed HCC cells, Aurora-A inhibitor has a higher efficacy in inhibiting cell growth. In HCC patients with high Aurora-A and Maf1 expression, treatment with Aurora-A inhibitors can be a promising strategy in cancer therapy. Citation Format: Ya-Ting Jhan, Yi-Seng Gan, Liang-Yi Hung. Aurora-a mediated phosphorylation of Maf1 is essential for maintaining protein stability and regulating tRNA biosynthesis in HCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1731.

Background: Colorectal cancer (CRC) progression and related mortality are highly associated with metabolic disorders. However, the molecular mechanism involved in the regulation of hyperlipidemia-associated CRC metastasis remains unclear. This study aimed to investigate the effects of angiopoietin-like 4 (ANGPTL4) on NADPH oxidase 4 (NOX4) expression and reactive oxygen species (ROS) production, which might provide new targets for improving outcomes in patients with hyperlipidemia-associated CRC metastasis. Methods: The clinical relevance of relationship between NOX4 expression and ANGPTL4 was examined in CRC patients by the Oncomine and TCGA data set. Expressions of NOX4, epithelial-mesenchymal transition (EMT) markers, and gene regulation of NOX4 in free fatty acids (FFAs)-treated CRC cells were determined. The FFAs-triggered metastatic ability of CRC cells under treatments of antioxidants or knockdown of NOX4, ANGPTL4, and MMPs was evaluated in vitro and in vivo. In addition, effects of antioxidants and depletion of metastasis-associated molecules on the correlation between ROS production and FFAs-promoted CRC metastasis were also clarified. Results: In this study, we found that the induction of NOX4, followed by the increased ROS was essential for oleic acid (OA)-promoted CRC cell metastasis. The depletion of ANGPTL4 significantly inhibited c-Jun-mediated transactivation of NOX4 expression, accompanied with reduced levels of ROS, MMP-1, and MMP-9, resulting in the disruption of OA-promoted CRC cell metastasis. Moreover, knockdown of ANGPTL4, NOX4, MMP-1, and MMP-9 or the treatment of antioxidants dramatically inhibited circulating OA-enhanced tumor cell extravasation and metastatic seeding of tumor cells in lungs, indicating that the ANGPTL4/NOX4 axis was critical for dyslipidemia-associated tumor metastasis. Conclusion: The coincident expression of NOX4 and ANGPTL4 in CRC tumor specimens provides the insight into the potential therapeutic targets for the treatment of dyslipidemia-associated CRC metastasis.

Angioimmunoblastic T-cell lymphoma (AITL) carries genetic mutations of TET2, RHOA, and IDH2, but the prognostic impact of these mutations is not widely investigated. Although one study shows no dif...

Centrosomal P4.1-associated protein (CPAP) is overexpressed in hepatocellular carcinoma (HCC) and positively correlated with recurrence and vascular invasion. Here, we found that CPAP plays an important role in HCC malignancies. Functional characterization indicated that CPAP overexpression increases tumor growth, angiogenesis, and metastasis ex vivo and in vivo. In addition, overexpressed CPAP contributes to sorafenib resistance. Mechanical investigation showed that the expression level of CPAP is positively correlated with activated STAT3 in HCC. CPAP acts as a transcriptional coactivator of STAT3 by directly binding with STAT3. Interrupting the interaction between CPAP and STAT3 attenuates STAT3-mediated tumor growth and angiogenesis. Overexpression of CPAP upregulates several STAT3 target genes such as IL-8 and CD44 that are involved in angiogenesis, and CPAP mRNA expression is positively correlated with the levels of both mRNAs in HCC. Knocked-down expression of CPAP impairs IL-6-mediated STAT3 activation, target gene expression, cell migration, and invasion abilities. IL-6/STAT3-mediated angiogenesis is significantly increased by CPAP overexpression and can be blocked by decreased expression of IL-8. Our findings not only shed light on the importance of CPAP in HCC malignancies, but also provide potential therapeutic strategies for inhibiting the angiogenesis pathway and treating metastatic HCC.

This study examined a high-performance film-type thermal interface material (F-TIM) with vertically aligned carbon nanofibers (VACNFs). Unlike conventional TIMs, which are composed of stacked, randomly oriented fillers of varying size, silicone-based F-TIMs with VACNFs provide high z-axis thermal conductivity and are highly compressible, with the ability to conform to surfaces of varying roughness. Thus, the heat transfer performance and bond line thickness (BLT) of the package can be controlled accordingly. In the present study, the maximum thermal conductivity of the F-TIM was 40 W/mK. The test vehicle was a 50 mm × 50 mm package that bonded with a 16 mm × 15 mm thermal testing die with a 150-W power input. Experiments revealed that the overall thermal resistance of a 100 μm-thick F-TIM was less than 0.036 °C/W. In contrast to the thermal resistance of a typical TIM, that of the VACNFs could be reduced by approximately 60%. Thermal cycling and high-temperature stability tests did not degrade the VACNFs, indicating that the F-TIM had excellent thermal stability.

Colorectal cancer (CRC) is a major health problem due to its high mortality rate. The incidence of CRC is increasing in young individuals. Oxaliplatin (OXA) is an approved third-generation drug and is used for first-line chemotherapy in CRC. Although current standard chemotherapy improves the overall survival of CRC patients, an increasing number of reports of OXA resistance in CRC therapy indicates that resistance has become an urgent problem in clinical applications. Dicer is a critical enzyme involved in miRNA maturation. The expression of Dicer has been reported to be involved in the resistance to various drugs in cancer. In the present study, we aimed to investigate the role of Dicer in OXA resistance in CRC. We found that OXA treatment inhibited Dicer expression through decreasing the protein stability. OXA-induced Dicer protein degradation occurred through both proteasomal and lysosomal proteolysis, while the CHIP E3 ligase was involved in OXA-mediated Dicer ubiquitination and degradation. We established stable OXA-resistant clones from CRC cells, and observed that the CHIP E3 ligase was decreased, along with the increased Dicer expression in OXA-resistant cells. Knockdown of Dicer resensitized CRC cells to OXA treatment. In this study, we have revealed the role of miRNA biogenesis factors in OXA resistance in CRC cells.

Angioimmunoblastic T-cell lymphoma (AITL) carries genetic mutations of

Many Aurora-A inhibitors have been developed for cancer therapy; however, the specificity and safety of Aurora-A inhibitors remain uncertain. The Aurora-A mRNA yields nine different 5'-UTR isoforms, which result from mRNA alternative splicing. Interestingly, we found that the exon 2-containing Aurora-A mRNA isoforms are predominantly expressed in cancer cell lines as well as human colorectal cancer tissues, making the Aurora-A mRNA exon 2 a promising treatment target in Aurora-A-overexpressing cancers. In this study, a selective siRNA, siRNA-2, which targets Aurora-A mRNA exon 2, was designed to translationally inhibit the expression of Aurora-A in cancer cells but not normal cells; locked nucleic acid (LNA)-modified siRNA-2 showed improved efficacy in inhibiting Aurora-A mRNA translation and tumor growth. Xenograft animal models combined with noninvasion in vivo imaging system (IVIS) analysis further confirmed the anticancer effect of LNA-siRNA-2 with improved efficiency and safety and reduced side effects. Mice orthotopically injected with colorectal cancer cells, LNA-siRNA-2 treatment not only inhibited the tumor growth but also blocked liver and lung metastasis. The results of our study suggest that LNA-siRNA-2 has the potential to be a novel therapeutic agent for cancer treatment.

// Yu-Chuan Huang 1 , Chung-Ta Lee 5, * , Jenq-Chang Lee 7, * , Yao-Wen Liu 8 , Ying-Jen Chen 3 , Joseph T. Tseng 1, 2 , Jui-Wen Kang 6 , Bor-Shyang Sheu 6 , Bo-Wen Lin 7 , Liang-Yi Hung 1, 2, 4, 9 1 Institute of Bioinformatics and Biosignal Transduction, National Cheng-Kung University, Tainan 70101, Taiwan 2 Department of Biotechnology and Bioindustry Sciences, National Cheng-Kung University, Tainan 70101, Taiwan 3 Department of Life Sciences, College of Bioscience and Biotechnology, National Cheng-Kung University, Tainan 70101, Taiwan 4 Center for Infectious Disease and Signal Transduction Research, College of Medicine, National Cheng-Kung University, Tainan 70101, Taiwan 5 Department of Pathology, National Cheng-Kung University Hospital, Tainan 70403, Taiwan 6 Department of Internal Medicine, National Cheng-Kung University Hospital, Tainan 70403, Taiwan 7 Department of Surgery, National Cheng-Kung University Hospital, Tainan 70403, Taiwan 8 Department of Pathology, Kuo General Hospital, Tainan 70054, Taiwan 9 Institute for Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan * These authors contributed equally to this work Correspondence to: Liang-Yi Hung, email: lyhung@mail.ncku.edu.tw Bo-Wen Lin, email: wen276@gmail.com Keywords: miR-137, Aurora-A, COX2, colorectal cancer, epigenetic regulation Received: May 14, 2016 Accepted: October 12, 2016 Published: October 18, 2016 ABSTRACT MicorRNA-137 is silenced in human colorectal cancer tissues and colon polyps. Our study showed that the decreased expression of miR-137 is significantly different in various types of polyp which maintain different potentials to lead to CRC development. The expression of miR-137 gradually decreases during the process of colorectal carcinogenesis. Receiver operating characteristic curve (ROC) analysis indicates that the loss of miR-137 expression in colon polyps can serve as a biomarker to predict the predisposition of colorectal carcinogenesis. By cell model and xenograft animal model, the enforced expression of miR-137 in colorectal cancer cells can inhibit cell proliferation and tumor formation, induce G2/M arrest, and lead to apoptosis. The expression pattern of miR-137 and Aurora-A or PTGS2 is negatively correlated in human colorectal cancer tissues and colon polyps. Those effects induced by overexpressed miR-137 can be rescued by the overexpression of Aurora-A. In summary, our study suggests that the loss of miR-137 expression in colon polyps can serve as a biomarker to predict the tendency toward to CRC formation through the impaired inhibitory effect of Aurora-A. The investigation of the regulatory mechanism of miR-137 -mediated Aurora-A inhibition may shed new light on the early prognosis of cancer therapy for CRC in the future.

Sclerosing angiomatoid nodular transformation (SANT) of the spleen is a morphologically distinctive lesion. Although the clinical course of SANT is benign, its reactive or neoplastic nature remains to be clarified. Furthermore, some investigators have suggested that SANT is related to IgG4 sclerosing lesion or inflammatory pseudotumor with stromal cells positive for Epstein-Barr virus (EBV). In this study, we assessed 22 cases of SANT derived from adult women. Clinical data and follow-up information were obtained by chart review. Immunohistochemical studies for IgG4, IgG, and CD21 stains and in situ hybridization to detect EBV-encoded small RNAs were performed. We also assessed genomic DNA extracted from paraffin-embedded tissue for human androgen-receptor α gene analysis using conventional and methylation-specific polymerase chain reaction methods. The median patient age was 41.5 years (range, 25 to 82 y). Most (77%) patients presented with a single mass that was detected incidentally (59%). The mean size of the lesions was 3.8 cm (range, 1.0 to 9.0 cm). Clinical symptoms correlated with multiple lesions (P=0.043) but not lesional size (P=0.637) or location in the spleen (hilum vs. periphery, P=0.696). None of the cases had evidence of IgG4-related disease or recurred after splenectomy. The mean number of IgG4 cells was 27.7 (range, 4 to 125), and the mean IgG4/IgG ratio was 16.4% (range, 1.6% to 55.7%) with only 2 cases being >40%. Cases with higher IgG4 cells did not correlate with inflammatory pseudotumor-like morphology. No lesions were positive for EBV-encoded small RNAs, and almost all cases with informative results (n=19) showed a polyclonal pattern. We conclude that SANT is a polyclonal, reactive lesion rather than a neoplasm.

Figure S1. HBx increases CPAP promoter activity in HCC cell lines. Figure S2. Expression of CPAP is increased in HBV genome-expressing HCC cells. Figure S3. The CREB binding site is crucial for CPAP promoter activity. Figure S4. CREB is essential for HBx-mediated CPAP promoter activity. Figure S5. Evaluation of CPAP promoter activity in shCREB transfected Hep3B cells. Figure S6. CPAP is involved in HBx-induced transcriptional activation of NF-κB. Figure S7. The interaction between CPAP and HBx is increased upon TNF-α treatment. Figure S8. CPAP promotes proliferation, colony formation, and tumorigenicity of HCC. Figure S9. NF-κB/p65 is essential for CPAP-mediated colony formation of HCC cells. Figure S10. Overexpression of CPAP/WT significantly increased tumor growth in a xenograft animal model. Figure S11. CPAP increases TNF-α-mediated HBx protein stabilization. Figure S12. The clinical outcome of overexpressed CPAP, HBx and activated NF-κB (p65) in HCC. Figure S13. Co-overexpression of CPAP and CREB is positively correlated with a poor disease-free survival rate in HBx-positive HCC. (PDF 1009 kb)

Hepatitis B virus (HBV) encoded non-structure protein X (HBx) can promote cell proliferation, migration, and anti-apoptosis via activating several transcription factors and increasing their downstream gene expression in HBV-infected liver cells. Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for HBx-mediated NF-κB activation. Here, we found that, upon HBV infection, overexpressed HBx can transcriptionally up-regulateCPAPvia interacting with CREB. CPAP can directly interact with HBx to promote HBx-mediated cell proliferation and migration; and SUMO modification of CPAP is involved in interacting with HBx. Interestingly, CPAP can increase the HBx protein stability in an NF-κB-dependent manner; and overexpressed CPAP and HBx is positively correlated with the activation status of NF-κB in HCC. Increased expression ofCREBandCPAPmRNAs exists in the high-risk group with a lower survival rate in hepatocellular carcinoma (HCC). These results suggest that the reciprocal regulation between CPAP and HBx may provide a microenvironment to facilitate HCC development via enhancing NF-κB activation, inflammatory cytokine production, and cancer maligancies. The findings of this study not only shed light on the role of CPAP in HBV-associated HCC, but also provide CPAP as a potential target for HBV-related HCC therapy.Author SummaryIn this study, we address a novel molecular mechanism for the collaboration between overexpressed HBx and CPAP in promoting hepatocarcinogenesis in HBV-associated HCC. Upon HBV infection, HBx is overexpressed and interacts with CREB to transcriptionally activate CPAP; the HBx/CPAP interaction promotes hepatocarcinogenesis. Clinical analysis found that co-overexpressed CPAP and CREB exist in the high-risk group with a lower survival rate in HCC. Additionally, overexpressed CPAP contributes to HBx protein stability in a NF-κB-dependent pathway. Our study provides a potential translational application in targeting CREB-CPAP axis in HBV-associated HCC.

Chemotherapy has a good success rate in the treatment of colorectal cancer; however, recurrence of colorectal cancer is still frequent due to acquired resistance. Aurora-A, a cell cycle-regulated kinase, plays an important role in colorectal cancer development and drug resistance. Dicer, one of the key enzymes of the microRNA (miRNA) biogenesis pathway, may be involved in chemoresistance through regulating the expression of miRNAs. According to the TCGA database, the expression levels of Dicer and Aurora-A were increased in colorectal cancer, which may contribute to chemoresistance. Our results showed that the protein levels of Dicer and Aurora-A are increased in oxaliplatin resistant cell lines. Knocked-down expression of Dicer or Aurora-A can enhance drug sensitivity in oxaliplatin-resistant cells; in contrast, overexpression of Dicer or Aurora-A increased drug resistance in parental drug sensitive cells. By next-generation sequencing, we found that the expression of a set of miRNAs is increased in oxaliplatin resistant cells. Dicer overexpression enhances the expression of those miRNAs in drug sensitive parental cells. Clinical evaluation further confirmed the increased expression of those miRNA in the plasma of CRC patients with a poor response to oxaliplatin. Our results suggest that those miRNAs may act as biomarkers to predict oxaliplatin response in colorectal cancer by detecting their plasma miRNAs. The molecular mechanism of those miRNAs in regulating the drug response is currently under our investigation. Citation Format: Li Jyuan Lin, Liang-Yi Hung. Dicer overexpression contributes to chemoresistance in colorectal cancer via up-regulating a set of miRNAs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3016.

Colorectal cancer (CRC) is highly associated with metabolic diseases, such as obesity and diabetes. Elevated free fatty acids (FFAs) in cancer patients with metabolic disorders may be associated with cancer progression. In addition, increased levels of reactive oxygen species (ROS) are also observed in dyslipidemia which induces expression of NADPH oxidase 4 (NOX4) by FFAs in aorta, kidney, and white adipocyte in physiology condition. Although the expression of NOX4 has been also up-regulated in various cancer cell types, the correlation between up-regulation of NOX4 and dyslipidemia-regulated CRC metastasis remains unclear. Here, we found that oleic acid (OA) induced-angiopoietin-like 4 (ANGPTL4) expression was through the activation of PPARs pathways, resulting in promoting the metastasis of CRC. The recombinant protein of human ANGPTL4 rescued the OA-induced invasive ability in ANGPTL4 knockdown cells. It is worthy to note that the knockdown of ANGPTL4 not only significantly inhibited OA-induced NOX4 expression, but also reduced ROS production. In addition, the depletion of ROS by using NAC or knockdown of NOX4 inhibited OA-induced extravasation of CRC cells in vivo. OA-induced MMP1 and MMP9 expressions were also dependent on the expression of ANGPTL4 and NOX4 in CRC cells. The transcriptional activation of NOX4 gene by OA-induced ANGPTL4 was regulated by activation of c-Jun and dependent on AP-1 site of NOX4 promoter. These results reveal that OA-promoted CRC metastasis was through the activation of ANGPTL4/NOX4 axis, suggesting that ANGPTL4 and NOX4 may be potential therapeutic targets and diagnostic markers to improving outcomes for patients with CRC. Citation Format: Chih-Jie Shen, Yu-Han Liao, Jhih-Peng Tsai, Liang-Yi Hung, Wen-Chang Chang, Ben-Kuen Chen. Oleic acid-induced ANGPTL4 facilitates metastasis of human colorectal cancer via up-regulation of NOX4 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2855.

Centrosomal P4.1-associated protein (CPAP) is a centrosomal protein and can as a transcriptional coactivator of STAT5 and NF-κB in cancer. Our previous studies indicated that CPAP is overexpressed in tumor tissue and can increase TNFα-mediated NF-κB activation in HCC. Here, we demonstrated that overexpressed CPAP increases tumor growth, angiogenesis, as well as metastasis ex vivo and in vivo. We found that CPAP increases these malignant abilities of tumor cells are through IL-6/STAT3 signaling. We demonstrated that CPAP directly interacts with C-terminal domain of STAT3 to increase STAT3 activity. Overexpression of CPAP enhances IL-6/STAT3/IL-8-mediated angiogenesis and other metastatic genes expression such as CD44 and MCAM. These results indicated that CPAP leads to HCC malignancy and metastasis by increasing STAT3 activity through directly interaction. Clinically, CPAP positively correlates with IL-8 in HCC with vascular invasion; and also positively correlates with CD44 and MCAM in HCC tissues. In summary, our findings shed light on the importance of CPAP to act as a potential therapeutic target for inhibiting the IL-6/STAT3-mediated angiogenesis pathway and treating metastatic HCC. Citation Format: Ruo Yu Chen, Liang-Yi Hung. CPAP promotes HCC angiogenesis and metastasis via interacting with and enhancing STAT3 signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1109.

Heterogeneous nuclear ribonucleoprotein (hnRNP) Q1, an RNA-binding protein, has been reported as overexpressing in cancer cells. However, the role of hnRNP Q1 in tumorigenesis remains unclear. By using RNA-immunoprecipitation (IP) assay following next-generation sequencing, a group of cell cycle-related genes targeted by hnRNP Q1 were identified, including Aurora-A kinase. Overexpressed hnNRP Q1 can up-regulate Aurora-A and many spindle assembly checkpoint (SAC) proteins, but not alter their mRNA levels, through enhancing translational efficiency by interacting with the mRNA 5’-UTR. Ribosomal S6-IP and ribosomal profiling assay further confirmed the translational regulation of these mRNAs by hnRNP Q1. Stimulation with epidermal growth factor (EGF) enhances both binding between hnRNP Q1 and these mRNAs as well as the efficacy of the hnRNP Q1-induced translation of these mRNAs. EGF/hnRNP Q1-induced translation is mediated by the mTOR and ERK pathways. In addition, hnRNP Q1 overexpression is positively correlated with the levels of Aurora-A and these SAC genes in human colorectal cancer tissues. Taken together, our data suggest that hnRNP Q1 plays an important role in translationally regulating the expression of Aurora-A and a group of cell cycle-related genes, which increases cell proliferation and contributes to tumorigenesis in colorectal cancer. It may thereby contribute to tumorigenesis by translationally up-regulating these genes in colorectal cancer. Citation Format: Liang-Yi Hung. hnRNP Q1 promotes cell proliferation and tumorigenesis by translationally up-regulating cell cycle-related genes through the EGF pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4387.

Overexpression of Aurora kinases is largely observed in many cancers, including hematologic malignancies. In this study, we investigated the effects and molecular mechanisms of Aurora kinase inhibitors in acute lymphoblastic leukemia (ALL). Western blot analysis showed that both Aurora-A and Aurora-B are overexpressed in ALL cell lines and primary ALL cells. Both VE-465 and VX-680 effectively inhibited Aurora kinase activities in nine ALL cell lines, which exhibited different susceptibilities to the inhibitors. Cells sensitive to Aurora kinase inhibitors underwent apoptosis at an IC50 of ∼10-30 nM and displayed a phenotype of Aurora-A inhibition, whereas cells resistant to Aurora kinase inhibitors (with an IC50 more than 10 μM) accumulated polyploidy, which may have resulted from Aurora-B inhibition. Drug susceptibility of ALL cell lines was not correlated with the expression level or activation status of Aurora kinases. Interestingly, RS4;11 and MV4;11 cells, which contain the MLL-AF4 gene, were both sensitive to Aurora kinase-A inhibitors treatment. Complementary DNA (cDNA) microarray analysis suggested that CDKN1A might govern the drug responsiveness of ALL cell lines in a TP53-independent manner. Most importantly, primary ALL cells with MLL-AF4 and CDKN1A expression were sensitive to Aurora kinase inhibitors. Our study suggests CDKN1A could be a potential biomarker in determining the drug responsiveness of Aurora kinase inhibitors in ALL, particularly in MLL-AF4-positive patients.

Background & Aims Constitutive activation of NF-κB is an important event involved in chronic inflammation in hepatocellular carcinoma (HCC). CPAP, which plays important roles in centrosomal functions, was previously identified as the transcriptional co-activator of NF-κB. However, the molecular mechanism is unclear. The goal of this study was to investigate the role of CPAP in activating the NF-κB pathway in HCC. Methods SK-Hep1, HuH7, HepG2, HepG2X, Hep3B, and Hep3BX cells with CPAP overexpression or CPAP siRNA were used to evaluate activation of NF-κB under TNF-α stimulation by reporter assay, RT-PCR, Q-PCR, and Western blot analysis. In vivo SUMO modification of CPAP was demonstrated by an in situ PLA assay. Human HCC tissues were used to perform Q-PCR, Western blot, and IHC. Results CPAP siRNA abolished the interaction between IKKβ and NF-κB, whereas overexpression of CPAP enhanced this interaction and finally led to augmented NF-κB activation by increasing the phosphorylation of NF-κB. CPAP could enter nuclei by associating with NF-κB. Furthermore, CPAP was SUMO-1 modified upon TNF-α stimulus, and this is essential for its NF-κB co-activator activity. SUMO-1-deficient CPAP mutant lost its NF-κB co-activator activity and failed to enter nuclei. Importantly, SUMOylated CPAP could synergistically increase the HBx-induced NF-κB activity. Conclusions CPAP is essential for the recruitment of the IKK complex to inactivated NF-κB upon TNF-α treatment. Expression of CPAP was positively correlated with a poor prognosis in HBV-HCC. CPAP has the potential to serve as a therapeutic target for inflammation and inflammation-related diseases.

// Chung-Ta Lee 1 , Yu-Chuan Huang 2, * , Liang-Yi Hung 2, 3, * , Nan-Haw Chow 1 , Pei-Fang Su 4 , Chung-Liang Ho 1 , Hung-Wen Tsai 1 , Yi-Lin Chen 1 , Shao-Chieh Lin 4 , Bo-Wen Lin 5 , Peng-Chan Lin 6 and Jenq-Chang Lee 5 1 Department of Pathology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 70403, Taiwan 2 Institute of Bioinformatics and Biosignal Transduction, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan 3 Department of Biotechnology and Bioindustry Sciences, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan 4 Department of Statistics, College of Management, National Cheng Kung University, Tainan 70101, Taiwan 5 Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 70403, Taiwan 6 Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 70403, Taiwan * These authors have contributed equally to this work Correspondence to: Chung-Ta Lee, email: lcta@mail.ncku.edu.tw Jenq-Chang Lee, email: leejc@mail.ncku.edu.tw Keywords: colorectal cancer, mucinous adenocarcinoma, serrated adenocarcinoma, CIMP, BRAF Received: December 14, 2016 Accepted: March 19, 2017 Published: April 04, 2017 ABSTRACT Colorectal mucinous adenocarcinoma (MAC) and serrated adenocarcinoma (SAC) share many characteristics, including right-side colon location, frequent mucin production, and various molecular features. This study examined the frequency of SAC morphology in MACs. We assessed the correlation of SAC morphology with clinicopathological parameters, molecular characteristics, and patient prognosis. Eighty-eight colorectal MACs were collected and reviewed for SAC morphology according to Makinen’s criteria. We sequenced KRAS and BRAF , assessed CpG island methylator phenotype (CIMP) frequency, and analyzed DNA mismatch repair enzyme levels using immunohistochemistry in tumor samples. SAC morphology was observed in 38% of MACs, and was associated with proximal location ( P =0.001), BRAF mutation ( P =0.042), CIMP-positive status ( P =0.023), and contiguous traditional serrated adenoma ( P =0.019). Multivariate analysis revealed that MACs without both SAC morphology and CIMP-positive status exhibited 3.955 times greater risk of cancer relapse than MACs having both characteristics or either one ( P =0.035). Our results show that two MAC groups with distinct features can be identified using Makinen’s criteria, and suggest a favorable prognostic role for the serrated neoplastic pathway in colorectal MAC.

By using RNA-immunoprecipitation assay following next-generation sequencing, a group of cell cycle-related genes targeted by hnRNP Q1 were identified, including Aurora-A kinase. Overexpressed hnRNP Q1 can upregulate Aurora-A protein, but not alter the mRNA level, through enhancing the translational efficiency of Aurora-A mRNA, either in a cap-dependent or -independent manner, by interacting with the 5′-UTR of Aurora-A mRNA through its RNA-binding domains (RBDs) 2 and 3. By ribosomal profiling assay further confirmed the translational regulation of Aurora-A mRNA by hnRNP Q1. Overexpression of hnRNP Q1 promotes cell proliferation and tumor growth. HnRNP Q1/ΔRBD23-truncated mutant, which loses the binding ability and translational regulation of Aurora-A mRNA, has no effect on promoting tumor growth. The expression level of hnRNP Q1 is positively correlated with Aurora-A in colorectal cancer. Taken together, our data indicate that hnRNP Q1 is a novel trans-acting factor that binds to Aurora-A mRNA 5′-UTRs and regulates its translation, which increases cell proliferation and contributes to tumorigenesis in colorectal cancer.

Background: The nucleolar MSP58 protein is a candidate oncogene implicated in cellular transformation. Results: MSP58 is associated with BRG1 and induces cellular senescence through the p53/p21 pathway. Conclusion: MSP58 has both tumor-suppressing and -promoting functions. Significance: This work reveals a novel role for MSP58 in cellular senescence and suggests that MSP58 may have further prognostic and therapeutic implications., The nucleolar 58-kDa microspherule protein (MSP58) protein is a candidate oncogene implicated in modulating cellular proliferation and malignant transformation. In this study, we show that knocking down MSP58 expression caused aneuploidy and led to apoptosis, whereas ectopic expression of MSP58 regulated cell proliferation in a context-dependent manner. Specifically, ectopic expression of MSP58 in normal human IMR90 and Hs68 diploid fibroblasts, the H184B5F5/M10 mammary epithelial cell line, HT1080 fibrosarcoma cells, primary mouse embryonic fibroblasts, and immortalized NIH3T3 fibroblasts resulted in induction of premature senescence, an enlarged and flattened cellular morphology, and increased senescence-associated β-galactosidase activity. MSP58-driven senescence was strictly dependent on the presence of functional p53 as revealed by the fact that normal cells with p53 knockdown by specific shRNA or cells with a mutated or functionally impaired p53 pathway were effective in bypassing MSP58-induced senescence. At least two senescence mechanisms are induced by MSP58. First, MSP58 activates the DNA damage response and p53/p21 signaling pathways. Second, MSP58, p53, and the SWI/SNF chromatin-remodeling subunit Brahma-related gene 1 (BRG1) form a ternary complex on the p21 promoter and collaborate to activate p21. Additionally, MSP58 protein levels increased in cells undergoing replicative senescence and stress-induced senescence. Notably, the results of analyzing expression levels of MSP58 between tumors and matched normal tissues showed significant changes (both up- and down-regulation) in its expression in various types of tumors. Our findings highlight new aspects of MSP58 in modulating cellular senescence and suggest that MSP58 has both oncogenic and tumor-suppressive properties.

Over-expression of AURKC has been detected in human colorectal cancers, thyroid carcinoma and several cancer cell lines. However, the regulation and clinical implications of over-expressed AURKC in cancer cells are unclear. Here we show that elevated AURKC increases the proliferation, transformation and migration of cancer cells. Importantly, the kinase activity of AURKC is required for these tumour-associated properties. Analysis of human cancer specimens shows that the expression of AURKC is increased in cervical cancer, and is highly correlated with staging in colorectal cancer. Over-expressed AURKC-GFP localizes to the centromeric regions of mitotic chromosomes and results in a decreased level of AURKB, a key regulator of spindle checkpoint. Expression of AURKC is down-regulated by PLZF, a transcriptional repressor, through recruitment to its promoter region. The expression levels of PLZF and AURKC mRNA display opposite patterns in human cervical and colorectal cancers. Taken together, our results provide important insights into human cancers with AURKC expression, which may serve as a potential target for cancer therapy in the future. Copyright  2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Chronic and persistent inflammation is a well-known carcinogenesis promoter. Hepatocellular carcinoma (HCC) is one of the most common inflammation-associated cancers; most HCC arises in the context of chronic inflammation and hepatic injury. Both NF-κB and STAT3 are important regulators for inflammation. Our previous studies indicated that CPAP (centrosomal P4.1-associated protein), a centrosomal protein that majorly participates in centrosome functions, is overexpressed in HCC and can increase TNFα-mediated NF-κB activation and IL-6-induced STAT3 activation. A hepatocyte-specific CPAP-expressing transgenic mouse was generated to investigate the physiological role of CPAP in hepatocarcinogenesis. An obvious inflammatory cell accumulation and fatty change were shown in the CPAP transgenic (Tg) mouse liver. The ALT level as well as the expression of inflammatory genes, such as IL-6, IL1β and TNF-α in CPAP Tg mouse are higher than in the wild type (WT) mouse. By high-dose and short-term diethylnitrosamine (DEN) treatment, the ALT level, pro-inflammatory genes and STAT3 activation are increased in CPAP Tg mouse; low-dose and long-term DEN treatment induces more severe liver tumor formation in CPAP Tg mouse than in WT mouse. In summary, these results suggest that CPAP may promote hepatocarcinogenesis through enhancing the inflammation pathway via activating the STAT3 pathway. Citation Format: Liang-Yi Hung. CPAP enhances diethylnitrosamine-induced hepatocarcinogenesis via STAT3 pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3086.

Loss of the maintenance of genetic material is a critical step leading to tumorigenesis. It was reported that overexpression of Aurora-A and the constitutive activation of the epidermal growth factor (EGF) receptor (EGFR) are implicated in chromosome instability. In this study, we examined that when cells treated with EGF result in centrosome amplification and microtubule disorder, which are critical for genetic instability. Interestingly, the expression of Aurora-A was also increased by EGF stimulus. An immunofluorescence assay indicated that EGF can induce the nuclear translocation of EGFR. Chromatin immunoprecipitation (ChIP) and re-ChIP assays showed significant EGF-induced recruitment of nuclear EGFR and signal transducer and activator of transcription 5 (STAT5) to the Aurora-A promoter. A co-immunoprecipitation assay further demonstrated that EGF induces nuclear interaction between EGFR and STAT5. A small interfering (si)RNA knockdown assay also showed that EGFR and STAT5 are indeed involved in EGF-increased Aurora-A gene expression. Altogether, this study proposes that the nuclear EGFR associates with STAT5 to bind and increase Aurora-A gene expression, which ultimately may lead to chromosome instability and tumorigenesis. The results also provide a novel linkage between the EGFR signaling pathway and overexpression of Aurora-A in tumorigenesis and chromosome instability.

Wogonin, a naturally occurring plant flavonoid, is isolated from Chinese herbal plants Scutellaria baicalensis Georgi and S. barbata D. Don. The extract of S. baicalensis Georgi has been added to an assortment of health drinks or food supplements. Wogonin has been reported to exhibit anticancer and anti-inflammatory properties. Cyclooxygenase-2 (COX-2) is a key enzyme in the production of prostaglandins in inflammatory conditions. In this study, the effect of wogonin on phorbol 12-myristate 13-acetate (PMA)-induced COX-2 expression was investigated. It showed that wogonin inhibited PMA-induced COX-2 protein and mRNA levels in human lung epithelial cancer cells, and the mechanism of this inhibition was at the transcriptional level by using COX-2 gene promoter assay. Among various signal inhibitors, the mitogen-activated protein kinase kinase 1/2 (MEK1/2) inhibitor U0126 also inhibited PMA-induced COX-2 expression and COX-2 promoter activation. The activity of AP-1-driven promoter, but not nuclear factor-kappa B (NF-kappaB), was inhibited by U0126. The data indicated that MEK1/2-AP-1 is very important for PMA-induced COX-2 expression. Wogonin also inhibited PMA-induced AP-1 activation and the expression of c-Jun, a key component of AP-1. Taken together, it is suggested that wogonin inhibits PMA-induced COX-2 gene expression by inhibiting c-Jun expression and AP-1 activation in A549 cells.

Epithelioid hemangioendothelioma (EHE) is a rare vascular tumor of low malignant potential that occurs mostly in soft tissue. So far, only two cases of EHE involving the intestine have been recorded. Here, we describe a rare case of cecal EHE due to a subsequent metastasis from the primary liver tumor three years after initial diagnosis. A 74- year-old man had sudden onset of epigastralgia for 24 hours. The abdominal CT revealed an ileocecal mass with a small bowel obstruction, and extensive tumor involvement in the liver was noted. He received an emergent right hemicolectomy for relief of the ileus. Unfortunately, he expired four days later due to septic shock. The pathologic diagnosis of EHE prompted a molecular study for a WWTR1-CAMTA1 fusion. The Sanger sequencing results showed the fusion involved exon 4 of WWTR1 with exon 8 of CAMTA1. There is no standard treatment for hepatic EHE because of its rarity and variable clinical outcome. The decision on a treatment strategy should be individualized for each patient. Since the patient received supportive care only for the liver tumor, this case demonstrated a natural course of hepatic EHE with a survival of more than 3.3 years.

The expression of cPLA2 is critical for transformed growth of non-small cell lung cancer (NSCLC). It is known that phorbol 12-myristate 13-acetate (PMA)-activated signal transduction pathway is thought to be involved in the oncogene action in NSCLC and enzymatic activation of cPLA2. However, the transcriptional regulation of cPLA2alpha in PMA-activated NSCLC is not clear. In this study, we found that PMA induced the mRNA level and protein expression of cPLA2alpha. In addition, two Sp1-binding sites of cPLA2alpha promoter were required for response to PMA and c-Jun overexpression. Small interfering RNA (siRNA) of c-Jun and nucleolin inhibited PMA induced the promoter activity and protein expression of cPLA2alpha. Furthermore, PMA stimulated the formation of c-Jun/Sp1 and c-Jun/nucleolin complexes as well as the binding of these transcription factor complexes to the cPLA2alpha promoter. Although Sp1-binding sites were required for the bindings of Sp1 and nucleolin to the promoter, the binding of nucleolin or Sp1 to the promoter was independent of each other. Our results revealed that c-Jun/nucleolin and c-Jun/Sp1 complexes play an important role in PMA-regulated cPLA2alpha gene expression. It is likely that nucleolin binding at place of Sp1 on gene promoter could also mediate the regulation of c-Jun/Sp1-activated genes.

SummaryCPAP is required for centriole elongation during S/G2 phase, but the role of CPAP in mitosis is incompletely understood. Here, we show that CPAP maintains spindle pole integrity through its phosphorylation by Aurora-A during mitosis. Depletion of CPAP induced a prolonged delay in mitosis, pericentriolar material (PCM) dispersion, and multiple mitotic abnormalities. Further studies demonstrated that CPAP directly interacts with and is phosphorylated by Aurora-A at serine 467 during mitosis. Interestingly, the dispersal of the PCM was effectively rescued by ectopic expression of wild-type CPAP or a phospho-mimic CPAP-S467D mutant, but not a non-phosphorylated CPAP-S467A mutant. Finally, we found that CPAP-S467D has a low affinity for microtubule binding but a high affinity for PCM proteins. Together, our results support a model wherein CPAP is required for proper mitotic progression, and phosphorylation of CPAP by Aurora-A is essential for maintaining spindle pole integrity.