Various Types Of Epithelial Cancer Research Articles

Predictive value of tumor budding in head and neck squamous cell carcinoma: an update.

Head and neck squamous cell carcinoma forms an anatomically and functionally complex group of malignancies. The significant local aggressiveness and frequent regional relapses motivate ongoing research to identify more reliable and sensitive prognostic and predictive biomarkers. One emerging area of cancer biology is the evaluation of tumor budding at the advancing invasive front of various types of epithelial cancers. Recent studies suggest that tumor budding is a relatively common phenomenon in cancer progression and that it may have important prognostic implications for patients due to its potential to provide valuable insights into the biology and clinical behavior of head and neck cancer. In this review, we aim to provide information about tumor budding in head and neck squamous cell carcinoma. Thus, we hope to shed light on the complex biology of these malignancies, as well as aiding diagnostic, classification, and better characterization and thereby, looking for new avenues for improving patient outcomes.

Cyclic AMP-Dependent Protein Kinase Exhibits Antagonistic Effects on the Replication Efficiency of Different Human Papillomavirus Types.

Several types of widespread human papillomaviruses (HPVs) may induce the transformation of infected cells, provoking the development of neoplasms. Two main genera of HPVs are classified as mucosatropic alphapapillomaviruses and cutaneotropic betapapillomaviruses (α- and β-HPVs, respectively), and they both include high-risk cancer-associated species. The absence of antiviral drugs has driven investigations into the details of the molecular mechanisms of the HPV life cycle. HPV replication depends on the viral helicase E1 and the transcription factor E2. Their biological activities are controlled by numerous cellular proteins, including protein kinases. Here, we report that ubiquitously expressed cyclic AMP-dependent protein kinase A (PKA) differentially regulates the replication of α-HPV11, α-HPV18, and β-HPV5. PKA stimulates the replication of both α-HPVs studied but has a more profound effect on the replication of high-risk α-HPV18. However, the replication of β-HPV5 is inhibited by activated PKA in human primary keratinocytes and U2OS cells. We show that the activation of PKA signaling by different pharmacological agents induces the rapid proteasomal degradation of the HPV5 E2 protein, which in turn leads to the downregulation of E2-dependent transcription. In contrast, PKA-stimulated induction of HPV18 replication is the result of the downregulation of the E8^E2 transcript encoding a potent viral transcriptional inhibitor together with the rapid upregulation of E1 and E2 protein levels. IMPORTANCE Several types of human papillomaviruses (HPVs) are causative agents of various types of epithelial cancers. Here, we report that ubiquitously expressed cyclic AMP-dependent protein kinase A (PKA) differentially regulates the replication of various types of HPVs during the initial amplification and maintenance phases of the viral life cycle. The replication of the skin cancer-related pathogen HPV5 is suppressed, whereas the replication of the cervical cancer-associated pathogen HPV18 is activated, in response to elevated PKA activity. To inhibit HPV5 replication, PKA targets the viral transcriptional activator E2, inducing its rapid proteasomal degradation. PKA-dependent stimulation of HPV18 replication relies on the downregulation of another E2 gene product, E8^E2, which encodes a potent transcriptional repressor. Our findings highlight, for the first time, protein kinase-related mechanistic differences in the regulation of the replication of mucosal and cutaneous HPV types.

Computational Evaluation of Bioactive Compounds from Colocasia affinis Schott as a Novel EGFR Inhibitor for Cancer Treatment.

Introduction:Epidermal growth factor receptor (EGFR) is a transmembrane protein that belongs to the ErbB/HER-family of tyrosine kinase receptors. Somatic mutations and overexpression of EGFR have been reported to play a vital role in cancer cell development and progression, including cell proliferation, differentiation, angiogenesis, apoptosis, and metastatic spread. Hence, EGFR is an important therapeutic target for the treatment of various types of epithelial cancers. Somatic mutations have led to resistance to clinically approved synthetic EGFR inhibitors. Furthermore, synthetic EGFR inhibitors have been associated with several side effects. Thus, there is a need to develop novel EGFR inhibitors with an acceptable biosafety profile and high efficacy.Methods:Herein, we employed structural bioinformatics and theoretical chemistry techniques via molecular docking, molecular mechanics generalized Born surface area (MM-GBSA) calculation, density functional theory analysis (DFT), and pharmacokinetic study to identify novel EGFR inhibitors.Results:The stringent molecular docking and MM-GBSA calculations identified MET 793, LYS 745, PHE 723, ASP 855, ARG 411, and THR 854 as principal amino acid residues for EGFR-ligands interactions. Furthermore, Colocasia affinis Schott compounds exhibited higher binding energy and more stable interactions than the reference compound (gefitinib). DFT analysis also ascertains better bioactivity and chemical reactivity of C. affinis Schott with favorable intramolecular charge transfer between electron-donor and electron acceptor groups. The pharmacokinetic profile of C. affinis Schott bioactive compounds satisfies Lipinski’s rule of five assessment.Conclusion:Collectively, C. affinis Schott compounds demonstrated higher inhibitory potentials against EGFR and better pharmacological properties when compared with gefitinib. C. affinis Schott compounds are therefore suggested as promising therapeutic EGFR inhibitors for cancer treatment.

Loss of HCRP1 leads to upregulation of PD-L1 via STAT3 activation and is of prognostic significance in EGFR-dependent cancer

Loss of hepatocellular carcinoma-related protein 1 (HCRP1) (alias VPS37A) plays a role in endocytosis of receptor tyrosine kinases as a member of the ESCRT complex and has been linked to poor patient outcome in various types of epithelial cancer. To this date, the molecular and biological mechanisms explaining how its absence would contribute to tumor progression remain unknown. Using genomic editing with CRISPR-Cas9, we generated ovarian and breast cancer cell lines with loss-of-function mutations of HCRP1. We hypothesized that pathways downstream of receptor tyrosine kinases such as epidermal growth factor receptor are affected by HCRP1 loss and looked for deregulated signaling using immunoblotting and classical cancer biology assays. In our study, we show that endogenous deletion of HCRP1 leads to elevated phosphorylation of the transcription factor Signal transducer and activator of transcription 3 (STAT3) and induces upregulation of PD-L1, an important regulator of immune checkpoint inhibition. HCRP1 loss further leads to a mesenchymal phenotype switch in cancer cells, leading to increased proliferation and migration. Concludingly, our data emphasize the role of the tumor microenvironment in tumors with low or absent HCRP1 expression and suggest HCRP1 loss as a potential marker for metastatic potential and immunogenicity of epidermal growth factor receptor-driven cancer.

Abstract PO-040: Oral squamous carcinoma cells under TGF-β-induced cell cycle arrest represent highly motile and invasive population

Abstract Cancer cells forming primary tumors are heterogeneous and respond in various ways to the stimuli from tumor microenvironment. Transforming growth factor-β (TGF-β) has been implicated in progression of various types of epithelial cancer. TGF-β is known to induce epithelial-mesenchymal transition (EMT) leading to increased invasiveness and thus metastasis. In addition, it has been reported that TGF-β arrests the epithelial cell in early G1 phase. While TGF-β elicits multiple effects on epithelial cancer cells, heterogeneity of tumor mass results in a different response depending on the cell state, thus it remains to be elucidated whether the cells exhibiting TGF-β-induced EMT and high motility are also under the cell cycle arrest. To examine the correlation between TGF-β-induced motility and cell cycle arrest at a single cell level, we have utilized oral squamous carcinoma cell lines, carrying fluorescent ubiquitination-based cell cycle indicator (Fucci) system. Using quantitative RT-PCR, immunoblotting and FACS analysis, we determined the expression of cell cycle-related genes and characterized the population of cells residing in certain cell cycle phases upon TGF-β treatment. Cells treated with TGF-β were also tested for their migratory ability to compare motility of cells residing in different cycle phases. Surprisingly, cells residing in G1 phase were characterized by higher motility than cells residing in S/G2/M phases suggesting a correlation between TGF-β-dependent cell cycle progression and migration. Moreover, increased motility of cells residing in G1 phase did not resulted from the progression of EMT. These results were also confirmed by our in vivo studies. In orthotropic xenograft model, cells under G1 arrest represented the majority of cells forming metastatic lymph nodules, in contrast to primary tumor mass comprising almost of equal number of cells residing in S/G2/M and G1 phases. We also performed cDNA microarray analyses of G1 and S/G2/M phase SAS-Fucci cells and identified gene whose expression was associated with both TGF-β-induced migration and cell cycle arrest. In conclusion our data suggested that cells under G1 cell cycle arrest are motile, but this increased motility is not a result of TGF-β-dependent EMT. Our data also strengthen the importance of targeting both, proliferating cancer cells and cell cycle-arrested cancer cells for the effective anti-cancer therapy. Citation Format: Katarzyna A. Inoue, Kazuki Takahashi, Maki Saito, Atsushi Kaida, Akinari Sugauchi, Toshihiro Uchihashi, Yasuhiro Yoshimatsu, Susumu Tanaka, Masahiko Miura, Mikihiko Kogo, Tetsuro Watabe. Oral squamous carcinoma cells under TGF-β-induced cell cycle arrest represent highly motile and invasive population [abstract]. In: Proceedings of the AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact; 2020 Sep 17-18. Philadelphia (PA): AACR; Cancer Res 2020;80(21 Suppl):Abstract nr PO-040.

Trop2 inhibition suppresses the proliferation and invasion of laryngeal carcinoma cells via the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway.

The cell surface glycoprotein Trop2 is overexpressed in various types of epithelial cancer. Laryngeal carcinoma is one of the most common types of head and neck cancer and in a previous study, the expression of Trop2 in laryngeal squamous cell carcinoma (LSCC) was identified as an independent prognostic factor. However, the biological significance of Trop2 in LSCC development remains unclear. In the current study, Trop2 protein expression in fresh LSCC tissue and paracancerous tissue was investigated using western blotting. Trop2 in the Hep2 laryngeal cell line was subsequently suppressed by transfection with small interfering RNA (siRNA). The effects of knockdown of Trop2 on cell viability, migration, invasiveness and ERK/MAPK pathway activity were investigated in the current study. The expression of Trop2 in fresh LSCC tissue was demonstrated to be significantly greater than that in paracancerous tissue. Trop2 expression was also identified to be required for proliferation, migration and invasiveness of Hep2 laryngeal carcinoma cells, as all were blocked by siRNA-mediated Trop2 inhibition. Notably, the ERK/MAPK signaling pathway and cell cycle factor, cyclin D1, were identified to be suppressed following the knockdown of Trop2 in Hep2 cells. These observations suggest that Trop2 serves an oncogenic role in LSCC and has potential as a therapeutic target.

Abstract P4-05-02: Role of the Atr Protein Kinase and Curcumin in Tumor-Stromal Interactions in Breast Cancer

Abstract Carcinoma-associated fibroblasts (CAFs) play important roles in the genesis and thrive of various types of epithelial cancers, including breast carcinomas. Indeed, various genetic and epigenetic variations have been identified in stromal fibroblasts, and we have recently shown that CAFs as well as their corresponding counterparts (TCFs) display neoplastic-specific changes (Hawsawi et al., 2008). We have also shown that the levels of the tumor suppressor proteins p53 and p21 are down-regulated in CAFs as compared to TCFs. During the cellular response to DNA damage these genes are under the control of important proteins such as Atr, a protein kinase essential for the maintenance of genomic integrity. To further elucidate the molecular changes that occur in breast cancer-associated fibroblasts we assessed the expression of Atr and have shown that the level of the Atr protein kinase is lower in 7 out of 10 (70%) CAFs as compared to their corresponding TCFs. Using specific ATR-siRNA we have shown that ATR negatively controls the expression of various proteins involved in the stromal-epithelial interactions. These include the stromal cell-derived factor 1 (SDF1), the vascular endothelial growth factor (VEGF) and the matrix metalloproteinase-2 (MMP2). In addition, the ELISA assay was used to show that the level of these proteins was higher in the conditioned media from stromal fibroblasts expressing specific ATR-siRNA as compared to control cells. Importantly, serum-free conditioned media from ATR-defective cells stimulated the proliferation and invasion/migration of cultured human breast cancer cells in an SDF1-dependent manner. These results clearly show the role of the breast stromal fibroblast Atr protein in suppressing tumoregenesis in the breast. Moreover, we have shown that curcumin can normalize the expression/secretion of these factors and therefore reduces the invasion/migration of breast cancer cells in vitro. This indicates that curcumin has potential use as stromal fibroblastnormalizing factor that can be utilized for the inhibition of both cancer initiation and recurrence. Hawsawi, N. M., Ghebeh, H., Hendrayani, S. F., Tulbah, A., Al-Eid, M., Al-Tweigeri, T., Ajarim, D., Alaiya, A., Dermime, S., and Aboussekhra, A. (2008). Cancer Res 68, 2717-2725. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-05-02.

Abstract 2328: Hyaluronan synthase expression in mammary tumor cells modulates tumor growth, angiogenesis and metastasis

Abstract Breast cancer is the second most common cause of cancer death in women in the United States after lung cancer and is responsible for more than 40,000 deaths each year. At the time of diagnosis, the majority of breast cancer patients have developed regional lymph node or distant metastases. Many challenges exist in the current management of metastatic breast cancer as there are fewer recognized therapeutic strategies. Therefore, a better understanding of the molecular events in the metastatic process is critical. Several reports have described correlation of hyaluronan (HA) with initiation and progression of various types of epithelial cancers. The HA synthase (HAS) isoforms encode the enzymes that produce and deposit HA while the hyaluronidase (HYAL) genes code for enzymes that degrade HA and their expression is dysregulated in various tumors as a result of transcriptional and epigenetic changes that accompany progression. In this report, we examined the expression of HAS1, HAS2 and HAS3, HYAL1 and HYAL2 in mammary tumor cell with different metastatic potential (4T1, highly metastatic; Cl66, moderately metastatic; Cl66M2, low metastatic). We observed increased expression of HAS1, HAS2 and HAS3 as well as HAYL 1 and HAYL2 in aggressive mammary tumor cells. Next we stably knocked-down HAS2 and HAS3 expression in 4T1 cells using small hairpin mRNA (sh-RNA) vectors and analyzed cell proliferation, migration, tumor growth and metastasis. We observed inhibition of in vitro cell proliferation and migration in 4T1 cells knocked-down for HAS2 (4T1-HAS2sh) and HAS3 (4T1-HAS3sh) as compared to cells transfected with vector control (4T1-controlsh). Furthermore, we observed inhibition of in vivo tumor growth and spontaneous lymph node and lung metastases in animals implanted with 4T1-HAS2sh cells as compared to 4T1-controlsh. In addition, we observed inhibition of tumor cell proliferation and neovascularization, and increased apoptosis in 4T1-shHAS2 tumors as compared to 4T1-controlsh tumors. Together, these data demonstrate an important role of HAS2 in mammary tumor growth, angiogenesis, invasion, and metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2328.

Combination of Cobe AutoPBSC and Gambro Elutra as a platform for monocyte enrichment in dendritic cell (DC) therapy: Clinical study

Monocytes are a common source for generating dendritic cells (DCs). The aim of the present study was to evaluate the efficiency of a platform for monocyte collection and enrichment in a clinical setting. The platform was based on the combination of two semiautomated devices; the Cobe Spectra Auto PBSC for mononuclear cells (MNC) collection followed by counterflow elutriation for monocyte enrichment (Gambro BCT Elutra). Twenty-four patients with various types of epithelial cancer participated in the study. MNC collections were first performed as large volume leukapheresis (LVL). Subsequently, MNC products were processed with an elutriation system for monocyte isolation. LVL resulted in the collection of MNC at a median of 8.1 x 10(9) cells, containing of 31.4% monocytes. A similar efficacy was also shown in patients with lower peripheral blood counts. Elutriation of the MNC product with the Cobe Elutra device resulted in the enrichment of monocytes at a median of 2.7 x 10(9) cells, with a recovery of 80.2% and a purity of 90.7%. These monocytes were then successfully developed into DCs for clinical therapy after in vitro manipulation. These data suggest that the combination of the Cobe Spectra Auto PBSC and the Gambro BCT Elutra is an effective platform for monocyte enrichment in clinical practice according to GCP standards and GMP guidelines, and can be easily implemented in the clinical routine under current DC protocols.

HER2/neu-derived peptides recognized by both cellular and humoral immune systems in HLA-A2+ cancer patients

HER2/neu is one of the most appropriate target antigens for anti-cancer therapy because of its expression in various types of epithelial cancer. HER2/neu can also be a target for both cellular and humoral immune responses. In this study, we attempted to identify HER2/neu-derived peptides, which are able to be recognized by both humoral and cellular immune systems in HLA-A2+ cancer patients. Among 12 HER2/neu-derived peptides having the HLA-A2 binding motifs, immunoglobulin G reactive to each of 7 HER2/neu peptides was detected in the plasma of >50% of cancer patients. Among these 7 peptides, 3 including HER2/neu 444-452, HER2/neu 466-474, and HER2/neu 484-492, effectively induced peptide-specific and HLA-A2-restricted cytotoxic T lymphocyte activity from peripheral blood mononuclear cells of cancer patients, regardless of different HLA-A2 subtypes. Experiments using blocking antibodies and cold inhibition targets revealed that the cytotoxicity against HER2/neu-expressing HLA-A2+ tumor cells was peptide-specific and CD8+ T cell-dependent. Overall, these results indicate that these 3 HER2/neu-derived peptides are efficiently recognized by both the humoral and cellular immune systems, and therefore could be useful for peptide-based immunotherapy for HLA-A2+ patients with various types of epithelial cancer.

Target molecules in specific immunotherapy against prostate cancer.

Recent advances in molecular biology and tumor immunology have allowed us to identify genes encoding human cancer-related antigens and their peptides that are recognized by cytotoxic T lymphocytes (CTLs). Although these advances have been preceded by studies on melanoma antigens, prostate cancer is another target candidate for specific immunotherapy. Several prostate tissue-specific antigens can be target molecules in specific immunotherapy for prostate cancer. The distribution of prostate tissue-specific antigens is more localized than that of melanoma-related antigens. Prostate-specific antigen (PSA) is available as an evaluation indicator of clinical course. In addition, epithelial cancer-related antigens are also applicable for prostate cancer patients. These lines of evidence suggest that prostate cancer is the best candidate for specific immunotherapy among the various types of epithelial cancers. A number of epitope peptides which have the potential to generate prostate cancer-reactive CTLs have been identified to date, and clinical trials targeting these molecules have been conducted. In this article, we review prostate cancer-related antigens and their epitope peptides, which have potential for use in the immunotherapy of prostate cancer patients, and we introduce the current status of clinical trials of specific immunotherapy targeting these molecules.