Differential molecular response of larynx cancer cell lines to combined VPA/CDDP treatment.

Successful treatment of advanced larynx squamous cell carcinoma (LSCC) remains a challenge, mainly due to limited response to chemotherapy and the phenomenon of the drug resistance. Therefore, new chemotherapeutic solutions are needed. The aim of this study was to explore benefit of combined cisplatin (CDDP) and valproic acid (VPA) therapy in patients' derived LSCC cell lines. Cell viability assay was used to establish cellular response to the drug by isobolography followed by RNA sequencing (RNAseq) analysis. Danio rerio were used for in vivo studies. Depending on the cell line, we found that the combinations of drugs resulted in synergistic or antagonistic pharmacological interaction, which was accompanied by significant changes in genes expression profiles. The presented therapeutic scheme efficiently blocked tumor growth in an in vivo model, corresponding to the in vitro performed studies. Interestingly the RK5 cell line, upon the combined treatment acquired a molecular profile typically associated with epithelial to mesenchymal transition (EMT). Hence, our studies demonstrates that patient-specific personalized therapy of larynx cancer should be considered and the combination of cisplatin and valproic acid should be explored as a potential therapeutic strategy in the treatment of larynx cancer.

Epidermal Growth Factor Receptor and its Oncogenic EGFRvIII Variant in Benign and Malignant Brain Tumors.

BACKGROUND/AIM: Tumorigenesis and cancer progression might be driven by abnormal activation of growth factor receptors. Importantly, molecular changes in EGFR-dependent signaling is one of the most common characteristics of brain tumors. PATIENTS AND METHODS: HER1 and EGFRvIII variants in meningiomas and glioblastomas were evaluated at the RNA level. RESULTS: EGFRvIII was found in 18.6% of glioblastomas (GBM), whereas 25% of EGFRvIII positive tumors express wild-type EGFR as well. HER1 was over-expressed in benign meningiomas compared to glioblastomas, whereas HER1 expression in meningiomas differed significantly between sub-types of meningiomas. EGFRvIII and HER1 where positively correlated in glioblastomas. Yet, the patient overall survival did not differ between high- and low-HER1 expressing glioblastomas or between EGFRvIII positive and negative GBMs. CONCLUSION: HER1 may be considered as an independent factor for classification of benign meningiomas. The mRNA levels of HER1 or EGFRvIII should not be used as independent prognostic factors for patients with gliomas.

Imperatorin as a Promising Chemotherapeutic Agent Against Human Larynx Cancer and Rhabdomyosarcoma Cells.

Naturally occurring coumarins are bioactive compounds widely used in Asian traditional medicine. They have been shown to inhibit proliferation, induce apoptosis, and/or enhance the cytotoxicity of currently used drugs against a variety of cancer cell types. The aim of our study was to examine the antiproliferative activity of different linear furanocoumarins on human rhabdomyosarcoma, lung, and larynx cancer cell lines, and dissolve their cellular mechanism of action. The coumarins were isolated from fruits of Angelica archangelica L. or Pastinaca sativa L., and separated using high-performance counter-current chromatography (HPCCC). The identity and purity of isolated compounds were confirmed by HPLC-DAD and NMR analyses. Cell viability and toxicity assessments were performed by means of methylthiazolyldiphenyl-tetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays, respectively. Induction of apoptosis and cell cycle progression were measured using flow cytometry analysis. qPCR method was applied to detect changes in gene expression. Linear furanocoumarins in a dose-dependent manner inhibited proliferation of cancer cells with diverse activity regarding compounds and cancer cell type specificity. Imperatorin (IMP) exhibited the most potent growth inhibitory effects against human rhabdomyosarcoma and larynx cancer cell lines owing to inhibition of the cell cycle progression connected with specific changes in gene expression, including CDKN1A. As there are no specific chemotherapy treatments dedicated to laryngeal squamous cell carcinoma and rhabdomyosarcoma, and IMP seems to be non-toxic for normal cells, our results could open a new direction in the search for effective anti-cancer agents.

Combination of Osthole and Cisplatin Against Rhabdomyosarcoma TE671 Cells Yielded Additive Pharmacologic Interaction by Means of Isobolographic Analysis.

BACKGROUND/AIM: Osthole is a simple coumarin that has been found to have anticancer, anti-inflammatory, antiviral, anticoagulant, anticonvulsant and antiallergic activities. The aim of this study was to analyze the combined anti-proliferative effect of cisplatin (CDDP) and osthole on a rhabdomyosarcoma cell line, and assess the pharmacology of drug-drug interaction between these drugs using isobolographic analysis. MATERIALS AND METHODS: The anticancer actions of osthole in combination with CDDP were evaluated using the tetrazolium dye-based MTT cell proliferation assay. RESULTS: Osthole and CDDP applied together augmented their anti-cancer activities and yielded an additive type of pharmacologic interaction by means of isobolographic analysis. CONCLUSION: Combined therapy using osthole and cisplatin could be suggested as a potential chemotherapy regimen against rhabdomyosarcoma.

Cyclooxygenase-2 Inhibition Enhances Proliferation of NKT Cells Derived from Patients with Laryngeal Cancer.

BACKGROUND/AIM: The aim of this study was to analyze whether inhibition of cyclooxygenase-2 by celecoxib and the subsequent enhancement in the proliferation of natural killer T (NKT) cells could play a role in dendritic cell (DC)-based laryngeal cancer (LC) immunotherapy. PATIENTS AND METHODS: Peripheral blood mononuclear cells were obtained from 48 male patients diagnosed with LC and 30 control patients without cancer disease. Neoplastic cell lysate preparations were made from cancer tissues obtained after surgery and used for in vitro DCs generation. NKT cells proliferation assay was performed based on 3H-thymidine incorporation assay. RESULTS: An increased proliferation of NKT cells was obtained from control patients compared to NKT cells obtained from LC patients regardless of the type of stimulation or treatment. In the patient group diagnosed with LC, COX-2 inhibition resulted in a significantly enhanced proliferation of NKT cells when stimulated with autologous DCs than NKT cells stimulated with DCs without COX-2 inhibition. These correlations were not present in the control group. Higher proliferation rate of NKT cells was also observed in non-metastatic and highly differentiated LC, which was independent of the type of stimulation or treatment. CONCLUSION: COX-2 inhibition could be regarded as immunotherapy-enhancing tool in patients with LC.

Additive Interaction of Cisplatin and Histone Deacetylase Inhibitors Combined Treatment in Rhabdomyosarcoma Cells - An Isobolographic Analysis.

BACKGROUND/AIM: The aim of this study was to assess the anticancer effect and the type of pharmacologic drug-drug interaction of cisplatin (CDDP) and histone deacetylase inhibitors (HDIs) combined treatment on the rhabdomyosarcoma cell line. MATERIALS AND METHODS: The antiproliferative actions of cisplatin and suberoylanilide hydroxamic acid (SAHA, vorinostat), as well as valproic acid (VPA) alone and in combination, were evaluated using the tetrazolium dye-based MTT cell proliferation assay and isobolographic analysis. RESULTS: All tested compounds inhibited proliferation of rhabdomyosarcoma cancer cells in a dose-dependent manner. The combinations of CDDP with SAHA and CDDP with VPA produced additive interaction with type-I isobolographic analysis. CONCLUSION: When adding SAHA or VPA to CDDP therapy, one can expect additive anticancer effects in the rhabdomyosarcoma cell line.

Histone Deacetylase Inhibitor SAHA as Potential Targeted Therapy Agent for Larynx Cancer Cells.

Objective: Laryngeal squamous cell carcinoma is one of the most common malignant tumors in the head and neck region. Due to the poor response to chemotherapeutics in patients and low survival rate, successful treatment of larynx cancer still remains a challenge. Therefore, the identification of novel treatment options is needed. We investigated the anticancer effects of suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, on two different laryngeal cancer cell lines RK33 and RK45. We also studied the antiproliferative action of SAHA in combination with cisplatin and defined the type of pharmacological interaction between these drugs. Materials and Methods: Viability and proliferation of larynx cancer cell lines were studied by methylthiazolyldiphenyl-tetrazolium bromide method and 5-bromo-2-deoxyuridine incorporation assay, respectively. The type of interaction between SAHA and cisplatin was determined by an isobolographic analysis. Western blotting, flow cytometry and quantitative polymerase chain reaction method were used to determine acetylation of histone H3, cell cycle progression and genes expression, respectively. Apoptosis was assessed by means of nucleosomes released to cytosol. Results: SAHA alone or in combination with cisplatin inhibited larynx cancer cells proliferation, whereas displayed relatively low toxicity against normal cells - primary cultures of human skin fibroblasts. The mixture of SAHA with cisplatin exerted additive and synergistic interaction in RK33 and RK45 cells, respectively. We showed that SAHA induced hyperacetylation of histone H3 K9, K14 and K23 and triggered apoptosis. SAHA also caused cell cycle arrest by upregulation of CDKN1A and downregulation of CCND1 encoding p21WAF1/CIP1 and cyclin D1 proteins, respectively. Conclusion: Our studies demonstrated that SAHA may be considered as a potential therapeutic agent against larynx tumors.

Isobolographic analysis demonstrates additive effect of cisplatin and HDIs combined treatment augmenting their anti-cancer activity in lung cancer cell lines.

Histone deacetylase inhibitors (HDIs) are a new class of drugs which affect the activity of HDACs resulting in changed of acetylation in many proteins. HDIs can induce differentiation, cell growth arrest, apoptosis, inhibit proliferation and angiogenesis in cancer, whereas normal cells are comparatively resistant to the action of HDIs. The aim of this study was to investigate the combined effect of a well-known cytostatic agent-cisplatin (CDDP) and a histone deacetylase inhibitors-either suberoylanilide hydroxamic acid (SAHA, vorinostat) or valproic acid (VPA), on the proliferation of lung cancer cells, as well as induction of apoptosis and inhibition of the cell cycle progression. The anti-proliferative activity of VPA or SAHA used alone, or in combination with CDDP were determined by means of MTT test. The type of pharmacologic interactions between HDAC inhibitors and CDDP was assessed using isobolographic analysis. We observed additive interactions for the CCDP with SAHA, as well as for the CDDP with VPA combinations with respect to their anti-proliferative effects on three different lung cancer cell lines (A549, NCI-H1563 and NCI-H2170). Such additive effects were observed regardless of the histologic type (adenocarcinoma or squamous cell carcinoma) and sensitivity for the drugs applied. Combination treatment also augmented the induction of apoptosis and cell cycle perturbation mediated by CDDP alone, thereby enhancing anti-cancer effect of tested drugs. In conclusion, the combined therapy of HDIs and CDDP may be a promising therapeutic tool in the treatment of lung cancer.

Assessment of Interactions between Cisplatin and Two Histone Deacetylase Inhibitors in MCF7, T47D and MDA-MB-231 Human Breast Cancer Cell Lines - An Isobolographic Analysis.

Histone deacetylase inhibitors (HDIs) are promising anticancer drugs, which inhibit proliferation of a wide variety of cancer cells including breast carcinoma cells. In the present study, we investigated the influence of valproic acid (VPA) and suberoylanilide hydroxamic acid (SAHA, vorinostat), alone or in combination with cisplatin (CDDP) on proliferation, induction of apoptosis and cell cycle progression in MCF7, T47D and MDA-MB-231 human breast carcinoma cell lines. The type of interaction between HDIs and CDDP was determined by an isobolographic analysis. The isobolographic analysis is a very precise and rigorous pharmacodynamic method, to determine the presence of synergism, addition or antagonism between different drugs with using variety of fixed dose ratios. Our experiments show that the combinations of CDDP with SAHA or VPA at a fixed-ratio of 1:1 exerted additive interaction in the viability of MCF7 cells, while in T47D cells there was a tendency to synergy. In contrast, sub-additive (antagonistic) interaction was observed for the combination of CDDP with VPA in MDA-MB-231 "triple-negative" (i.e. estrogen receptor negative, progesterone receptor negative, and HER-2 negative) human breast cancer cells, whereas combination of CDDP with SAHA in the same MDA-MB-231 cell line yielded additive interaction. Additionally, combined HDIs/CDDP treatment resulted in increase in apoptosis and cell cycle arrest in all tested breast cancer cell lines in comparison with a single therapy. In conclusion, the additive interaction of CDDP with SAHA or VPA suggests that HDIs could be combined with CDDP in order to optimize treatment regimen in some human breast cancers.

Osthole induces apoptosis, suppresses cell-cycle progression and proliferation of cancer cells.

BACKGROUND: The aim of the present study was to determine the effects of osthole on cell proliferation and viability, cell-cycle progression and induction of apoptosis in human laryngeal cancer RK33 and human medulloblastoma TE671 cell lines. MATERIALS AND METHODS: Cell viability was measured by means of the MTT method and cell proliferation by the 5-bromo-2-deoxyuridine (BrdU) incorporation assay. Cell-cycle progression was determined by flow cytometry, and induction of apoptosis by release of oligonucleosomes to the cytosol. The gene expression was estimated by a quantitative polymerase chain reaction (qPCR) method. High-performance counter-current chromatography (HPCCC) was applied for isolation of osthole from fruits of Mutellina purpurea. RESULTS: Osthole decreased proliferation and cell viability of cancer cells in a dose-dependent manner. The tested compound induced apoptosis, increased the cell numbers in G1 and decreased cell number in S/G2 phases of the cell cycle, differentially regulating CDKN1A and TP53 gene expression depending on cancer cell type. CONCLUSION: Osthole could be considered as a potential compound for cancer therapy and chemoprevention.

MicroRNAs-role in lung cancer.

Regulation of gene expression is essential for normal physiological functions; thus deregulation of gene expression is common in disease conditions. One level of regulation of gene expression is performed by noncoding RNAs, among which microRNAs (miRNA) are the best studied. Abnormal expression of these molecular players can lead to pathogenic processes such as heart disease, immune system abnormalities, and carcinogenesis, to name but a few. Of a length of 18-25 nucleotides miRNAs are involved in binding partial complementary sequences within the 3'-UTR (3'-untranslated region) of the target mRNAs. Depending on the type of neoplastic transformation, miRNAs can act both as oncogenes (oncomirs) or as tumor suppressors. Because of the great importance of miRNAs, most researches focus on either their role as biomarkers or their potential as therapeutic targets. Herein, we present the review of microRNA biology, function, and tumorigenic potential with emphasis on their role in lung cancer.

Glutamate and its receptors in cancer.

Glutamate, a nonessential amino acid, is a major bioenergetic substrate for proliferating normal and neoplastic cells on one hand and an excitatory neurotransmitter that is actively involved in biosynthetic, bioenergetic, metabolic, and oncogenic signaling pathways on the other. It exerts its action through a family of receptors consisting of metabotropic glutamate receptors (mGluRs) and ionotropic glutamate receptors (iGluRs), both of which have been implicated previously in a broad spectrum of acute and chronic neurodegenerative diseases. In this review, we discuss existing data on the role of glutamate as a growth factor for neoplastic cells, the expression of glutamate receptors in various types of benign and malignant neoplasms, and the potential roles that GluRs play in cancer development and progression along with their clinical significance. We conclude that glutamate-related receptors and their signaling pathways may provide novel therapeutic opportunities for a variety of malignant human diseases.

7-substituted coumarins inhibit proliferation and migration of laryngeal cancer cells in vitro.

BACKGROUND: Coumarins are a large group of naturally-occurring compounds with a wide range of biological properties, including anticancer activity. 7-Substituted coumarins (umbelliferone, scoparone, and herniarin) were analyzed for their potential anticancer activity against laryngeal cancer cells (LCC). MATERIALS AND METHODS: High-performance counter-current chromatography was applied for successful separation of umbelliferone from fruits of Heracleum leskowii. A two-phase solvent system composed of n-heptane-methanol-ethyl acetate-water (1:2:1:2, v/v/v) was successfully used. Cell proliferation was assessed after 48-72 h by means of MTT test, and tumor cell motility by a wound assay model. Measurement of cell death was estimated using enzyme-linked immunosorbent assay (ELISA), and cell-cycle analysis was performed by flow cytometry. Extracellular signal-regulated kinases-1/2 (ERK1/2) and AKT kinase activation status were analyzed by western blotting. RESULTS: Umbelliferone, scoparone, and, to a lesser extent, herniarin reduced viability and migration of RK33 LCC in a dose-dependent manner. Scoparone and herniarin were found to induce apoptosis of LCC. None of the tested compounds influenced the ERK1/2 and AKT kinase activity, nor significantly affected cell-cycle progression in the LCC line studied. CONCLUSION: Our findings suggest the therapeutic potential of 7-substituted coumarins in the treatment of laryngeal cancer.

Glutamate receptors in laryngeal cancer cells.

AIM: Despite recent improvements in treatment strategies, the results of chemotherapy in patients with advanced squamous cell carcinoma of the larynx are not satisfactory. Thus, the development of new approaches which influence specific metabolic pathways are needed. In the last decade, evidence has emerged implicating a role for glutamate as a signal mediator in tumors. MATERIALS AND METHODS: The presence of glutamate receptor subunits in two laryngeal cancer cell lines (RK33 and RK45) was evaluated by means of end-point PCR, real-time PCR, and immunocytochemistry. RESULTS: Glutamate receptor subunits are differentially expressed in laryngeal cancer cell lines. In addition, we show that selected ionotropic glutamate receptor antagonists and metabotropic glutamate receptor 5 antagonist inhibit proliferation of laryngeal cancer cells. Glutamate antagonists also affected activity of extracellular signal-regulated kinases 1/2 in tumor cells. CONCLUSION: Signaling through glutamate receptors may influence growth of laryngeal cancer cells and may constitute an adjunctive therapeutic target.

[The case of pleomorphic adenoma in inferior nasal concha]. / Przypadek gruczolaka wielopostaciowego malzowiny nosowej dolnej.

The authors present the case of inferior nasal concha tumor at early growth stage. The tumor which turned out to be pleomorphic adenoma was detected accidentally at 28 years old patient with a nose injury. This localisation of pleomorphic adenoma is extremely rare. The diagnostics, description of surgery and result of a histopathological examination were presented. Reports of a literature of pleomorphic tumors located in atypical places were quoted. Authors emphasize unanimous surgical treatment and necessity of observation after surgery because of tumor recurrence possibility.

[Lack of prognostic significance of carcinoembryonic antigen (CEA) preoperative serum levels in patients with advanced larynx cancer]. / Brak wartosci diagnostycznej przedoperacyjnego oznaczania steienia antygenu rakowo-plodowego (CEA) w surowicy pacjentów z zaawansowanym rakiem krtani.

The purpose of this study was to identify the relationship between preoperative serum levels of carcinoembryonic antigen (CEA) and clinicopathological features in advanced stage of larynx cancer. The mean CEA serum concentrations were below cutoff value, which exclude the CEA as a serum marker in diagnosis of larynx cancer. However, significant correlations were found between CEA levels and tumor size, lymph node metastasis and clinical stage of the disease. The pretreatment CEA level was increased above cut-off value only in 6% of tested patients and thereby is not a prognostic factor in larynx cancer.

[Diagnostic value of tissue polypeptide antigen in serum of larynx cancer patients]. / Wartosc diagnostyczna oznaczania tkankowego antygenu polipeptydowego w surowicy chorych na raka krtani.

UNLABELLED: Tissue polypeptide antigen (TPA) is a serological tumour marker used in the diagnosis, management and follow-up of patients with head and neck cancer. The aim of this study was to evaluate the diagnostic and prognostic significance of pretreatment TPA serum levels in patients with larynx cancer. The predicting ability of this tumour marker with respect to histological type, pathological state and lymph node metastasis was also assessed. MATERIALS AND METHOD: Concentrations of the TPA in the serum from 35 patients were measured by immunoradiometric assay. RESULTS: The results showed the sensitivity value for the group of 35 patients was 29.4%, but in clinical stage IV was 70%. TPA levels correlate with T classification and lymph node metastasis. In T4 tumors it was significantly higher than in T2 (p=0.047). TPA levels were significantly higher in patients with nodal invasion and were generally lower in patients with spinocellular carcinoma (p=0.0048). CONCLUSION: Our date indicate that TPA is of limited usefulness in the primary diagnosis in patients with larynx cancer, but is useful in detecting lymph node metastasis.

[Laser Nd-Yag assisted myringotomy for secretory otitis media in children]. / Myringotomia z zastosowaniem lasera Nd-YAG w leczeniu wysikowego zapalenia ucha srodkowego u dzieci.

The aim of the study was assessment of the qualities of ND-YAG laser assisted myringotomy as a treatment for secretory otitis media. Nd-Yag laser myringotomy was performed on 52 children (104 ears) mean age 6.8 years diagnosed with secretory otitis media and recurrent secretory otitis media (14.4%). Myringotomy was performed under general anesthesia using the laser (CWNd-YAG laser 3100 Sharplan). Adenoidectomy alone or with tonsillotomy was performed at the same time. Laser tympanostomies remained patent for 6-35 days. All tympanostomies healed with no noticeable scarring. In 15 ears pressure equalization tubes were inserted after myringotomy with laser. Septoplasty performed in 5 children. Laser myringotomy appears to be a safe, and easy to performed, alternative technique in the treatment of secretory otitis media.

[Chronic sinusitis therapy with antibiotics (axetyl cefuroxym, clarithromycin) and steroid (prednisone)]. / Leczenie zapalenia zatok z zastosowaniem antybiotyków (aksetyl cefuroksymu, klarytromycyna) w polaczeniu ze sterydem stosowanym doustnie (prednison).

We started therapy sinusitis of our patients with antibiotics cefuroxime axetil (Zinnat, GSK), clarithromycin (Klacid Uno, Abbott) and orally given steroid-prednisone in one group (A+S) 56 patients. Second group of 60 patients were cured only with antibiotics (A). We compare effects of this therapy. There were 50% totally cured patients in the first group (A+S) and 46.6% cured in the second group. Percentage totally cured patients with (A+S) is 3.4% better that cured only with antibiotics in the same time. It is statistically important. We present benefits for patients who were operated in the next step. Post therapy with the use of antibiotics and steroids there were less bleeding from the mucosal membrane, and there was no edema. It is a good method of therapy if patients have no contraindications.

[Squamous cell carcinoma antigen levels in patients with laryngeal cancer]. / Antygen raka plaskonablonkowego w surowicy krwi pacjentów z rakiem krtani.

Squamous cell carcinoma antigen (SCCAg) is one of the most common markers used in diagnosis of head and neck cancer and larynx cancer. We tested correlations between level of SCC Ag and tumor size, presence of lymph node metastasis, clinical advances of tumour and histopathological diagnosis. Pretreatment level of SCC antigen was evaluated in 34 patients with squamous cell carcinoma of the larynx. Microparticle enzyme immunoassay was used to measure the SCCAg level. Elevated SCCAg serum levels were found in 41% of patients. The magnitude of the marker elevations were correlated with lymph node metastases (N0 versus N2, and N1 versus N2). Our date indicate that in patients with larynx cancer SCCAg does not appear to be a sensitive marker in the primary diagnosis. However, seem to be useful marker for monitoring nodal invasion.