Non-small cell lung cancer (NSCLC) is one of the most lethal malignancies accountings for nearly 80% of all lung cancer cases diagnosed and causing over one million deaths annually worldwide. The discovery of molecular alterations including driver mutations and gene fusions has led to innovation of numerous targeted therapies, which certainly provided an edge over the classical chemotherapeutic treatment regimens and improved survival of the patients. Despite all the breakthrough innovations, the five-year survival statistics has not improved the way it was expected, pointing the challenges and limitations of currently approved diagnostic methods and therapies. This review summarizes various innovative therapies, treatment regimens developed over the last two decades for NSCLC treatment and the current challenges and limitations in the NSCLC treatment landscape.
BACKGROUND:Prostate cancer (PCa) is the second most frequently diagnosed cancer in males worldwide and placed fifth in cancer mortality among males. Between 14-24% of PCa patients have newly diagnosed advanced stages, which paradoxically has remained stable over time.AIM:To estimate and compare long-term radical prostatectomy (RP) oncologic outcomes in patients with clinically locally advanced prostate cancer (LAPCa), to determine the prognostic significance of common clinical-pathological parameters.PATIENTS AND METHODS:The study included 105 patients with LAPCa who underwent RP with extended pelvic lymphadenectomy between September 2003 - April 2015. Kaplan - Meier method was used for calculating biochemical recurrence- (BRFS), progression-free- (PFS), overall (OS), and prostate cancer-specific survival (PCSS) rates. Analyses of features associated with outcomes were conducted using Cox proportional hazards regression model.RESULTS:Patients from cT3b group had worse PFS, OS and PCSS rates in comparison with cT3a, while there was no significant difference in BRFS rates. Preoperative serum prostate-specific antigen level (hazard ratio (HR) 1.023, 95% confidence interval (CI): 1.014-1.033, p < 0.001), pT3a (HR 3,027, 95% CI: 1.449-7.096, p < 0.01), pT3b (HR 2.792, 95% CI: 1.133-6.881, p < 0.05) pT4 stage (HR 31.12, 95% CI: 7.646-126.6 p < 0.001) and positive lymph nodes status (HR 6.503, 95% CI: 3.190-13.25, p < 0.001) were significant factors in BRFS. Preoperative serum prostate-specific antigen level (HR 1.018, 95% CI: 1.007-1.030, p = 0.001) and positive lymph nodes status (HR 3.191, 95% CI: 1.672-6.088, p < 0.001) were significant factors in PFS and PCSS.CONCLUSIONS:RP as the initial treatment option of multimodal therapy in the management of LAPCa patients demonstrates encouraging oncologic outcomes. Patients from the cT3b group had the worse rates of PFS, OS, and PCSS in comparison with the cT3a group. Heterogeneity of LAPCa patients' outcomes reflects the insufficiency of the existing clinical risk classification for the prediction of systemic progression and cancer-specific survival.
BACKGROUND:Side effects of chemotherapy in cancer patients need to be investigated in more detail.AIM:To determine the incidence of cardiotoxicity in patients treated with different chemotherapy regimens containing 5-fluorouracil (5-FU) in Zanjan, Iran.PATIENTS AND METHODS:In a prospective cohort study, patients with different types of solid gastrointestinal tumors who were candidates for 5-FU based chemotherapy regimens were enrolled. The study population consisted of 100 patients (48 females and 52 males) with a mean age of 63.99 ± 12.40 years. We measured serum cardiac troponin I (cTnI) levels before and during each chemotherapy cycle and determined the occurrence of cardiotoxicity in patients based on the levels of cTnI, clinical signs and symptoms as well as electrocardiogram findings. In addition, we assessed a history of diabetes, hypertension, smoking, dyslipidemia and previous chest radiation as potential risk factors for cardiotoxicity.RESULTS:The incidence of cardiotoxicity was 8%, of which 5 patients were diagnosed with acute coronary syndrome, 2 patients with arrhythmias and one with hypotension. In addition, there was no significant association between studied risk factors and 5-FU induced cardiotoxicity.CONCLUSION:The incidence of cardiotoxicity in patients receiving 5-FU infusion regimens was notable. Thus, paying more attention to the 5-FU-induced cardiotoxicity is necessary in order to improve the prognosis of patients with cancer.
BACKGROUND Approximately 15% to 24% of essential thrombocythemia (ET) and 25-35% of primary myelofibrosis cases carry a mutation in the calreticulin (CALR) gene. Sanger sequencing, qPCR, high resolution melt or targeted next generation sequencing usually used to detect these mutations are expensive and require costly equipment. Nevertheless, type 1 CALR mutations are detectable by using polymerase chain reaction (PCR) and agarose gel electrophoresis. AIM To offer the use of the allele-specific reverse transcription (RT) PCR for rapid low-cost detection of the type 2 mutation in the CALR gene. MATERIALS AND METHODS Allele-specific primers designed for detecting type 2 mutation (5-bp insertion; c.1154_1155 ins TTGTC) of the CALR gene were used for allele-specific RT-PCR analysis of cDNA of the patient with JAK2-, MPL-negative ET, whose mutation in CALR gene has been identified by Sanger sequencing. RT-PCR samples were analyzed by agarose gel electrophoresis. RESULTS The type 2 mutation (K385fs*47 ins5) in CALR gene was detected by Sanger sequencing in JAK2- and MPL-negative ET patient. The cDNA obtained was then re-analyzed by using allele-specific RT-PCR with newly designed primers. Normal and type 2 mutation alleles of the CALR gene were detected by gel electrophoresis. The results of allele-specific RT-PCR were consistent with the data of Sanger sequencing. CONCLUSION Allele-specific RT-PCR analysis may be used for the fast low-cost detection of the major type 2 mutation (ins 5) of the CALR gene in patients with MPNs.
Background: G-force is a fundamental force controlling human cells. Cancer is one of the 4 major health challenges in the Space missions. Cancer in Space project evaluates the reaction of human cancer cells to the conditions of the space flights, including an exposure to high g-forces. Aim: Explore an impact of 10 g force on the oncogenic properties of human breast adenocarcinoma cells MCF-7. Materials and Methods: Cells were exposed to 10 g force for 10 days, as part of a 6-week simulation of conditions of a space flight. Then the cells were cultured for one week under normal culture conditions, before performing tests. Cell proliferation, cell viability, cell-cell contact inhibition, migration, and invasiveness were measured. Immunoblotting was used to evaluate expression of proteins. Results: Proliferation, cell-cell interaction and formation of 3D structures, migration, and invasiveness of cells exposed to 10 g were compared to parental cells cultured at 1 g condition. 10 g exposed cells showed a higher propensity for cell-cell contact inhibitions and lower for 3-dimensional growth in dense culture. This correlated with the decrease of proliferation in a dense culture as compared to the parental cells. The decrease of migration, adherence to a surface, and invasiveness was observed for cells subjected to the hypergravity, as compared to the parental MCF-7 cells. Enhanced expression of E-cadherin and phosphorylated pY576-FAK were observed in 10 g exposed cells but no impact on the expression of Erk, pErk, FAK and p53 was detected. Conclusion: The prolonged exposure of MCF-7 cells to 10 g force targets cell-cell and cell-substrate interactions.
AIM:To assess ultrasound (US) method informativeness in the recurrence detection and treatment monitoring of patients with cervical cancer metastases in the vagina via comparing the US and magnetic resonance imaging (MRI) data.MATERIALS AND METHODS:42 patients with recurrence of cervical cancer were examined by transvaginal US and MRI. Data on radiation diagnostics of cervical cancer metastasis in the vagina were compared with pathomorphological data. US and MRI data on the metastatic tumors size were compared.RESULTS:The diagnostic efficiency of US and MRI for recurrence detection was determined. The sensitivity, specificity and accuracy of US were 92.8; 93.3 and 93.3%, respectively, and of MRI - 95.2; 96.6 and 95.8%, respectively. The informativeness of MRI was higher than US in cervical cancer recurrences detection, but the difference was not significant (p > 0.05). US can provide data as informative as MRI for estimation of metastases sizes in the vagina. The mean difference between MRI measurements and US measurements of the metastases volume was 0.79 mm (95% CI 0.62-0.95 cm3).CONCLUSIONS:US as a cheaper and simpler method could be an alternative for metastases detecting and treatment monitoring, especially if there are contraindications to MRI.
AIM To determine the effect of 2-[(3-carboxy-1-oxoprogy1) amino]-2-deoxy-D-glucose (COPADG) on proliferation and apoptosis of human hepatocellular carcinoma cells (Hep G2). METHODS Hep G2 cells were cultured in RPMI-1640 medium in the presence of various concentrations of COPADG. Cell proliferation was determined by MTT assay. Apoptosis was determined by fluorescence microscopy, transmission electron microscopy, agarose gel electrophoresis of DNA fragmentation, and flow cytometry. RESULTS At the concentration ranging between 1-30 microM, COPADG potently inhibits growth and induce apoptosis of Hep G2 cells. CONCLUSIONS COPADG could effectively induce apoptosis in Hep G2 cells. It may be potentially useful as a new agent for treatment of human hepatocellular carcinoma.
In the review, the role of integrin receptors as well as of some of their ligands of the extracellular matrix in cases of parotid neoplasms is discussed, in particular, the data concerning the expression of various types of integrin adhesion molecules in benign and malignant tumors of the parotid, the expression of fibronectin, tenascin, and collagen type IV etc. The role of dysregulation of the expression of these proteins in invasion and metastasis of parotid tumors is discussed.
UNLABELLED CpG DNA are potent immunostimulator and currently being tested as adjuvant in immunotherapy of various diseases. THE AIM of this study was to investigate the effects of a single dose of bacterial CpG DNA challenge on response of murine immune system organs and on a zeta-potential of different lymphoid organ cells and peritoneal macrophages in normal and tumor-bearing mice. METHODS Indexes and cellularity of immune organs of mice were evaluated. Z-potential was measured by cellular electrophoresis. RESULTS Subcutaneous administration of CpG DNA induced local transitory hyperplasia of lymph nodes, moderate increase of splenic index and total spleen cellularity, whereas intraperitoneal injection of CpG DNA caused full-blown spleen enlargement and increase of splenocyte content, and also mice showed transitory aseptic peritonitis. Response of thymus on challenge of CpG DNA was bi-phase: initial phase - response on antigen as stimulus, and late proliferative phase. It was noted that tumor growth does not affect zeta-potential in peritoneal macrophages and mononuclear lymphocytes, but causes increase of zeta-potential on thymocytes and decreases it in lymphocytes from lymph nodes. Furthermore, single administration of CpG DNA normalizes of thymocytes and lymph nodes lymphocytes zeta-potential and increases it in peritoneal macrophages and mononuclear lymphocytes. CONCLUSION The findings demonstrate a close correlation between the hyperplasia of lymphoid follicles induced by challenge of CpG DNA and increase of their cellularity. Observed Z-potential alterations of immune system cells after CpG DNA immunization evidence on more significant polyanion accumulation on the surface of splenic macrophages and mononuclear cells.
AIM To develop a rationally designed new nitrogen mustard namely Fluorenhymustine (compound 2), where N,N'-bis(2chloro-ethyl)amino group, the established anticancer functionality, is attached to the 2-ethyl fluorenone hydantoin moiety. MATERIALS AND METHODS Starting from fluorenone hydantoin, a 3-step synthetic procedure was followed to obtain the title compound. 4-(4-Nitrobenzyl)pyridine was used to assess its chemical alkylating activity. Murine tumors (Ehrlich ascites carcinoma (EAC) and Sarcoma-180 (S-180)) were used to assess its in vivo activity. Its cytotoxicity was determined in vitro in MCF-7 human breast tumor cell line, toxicity - in vivo in normal and EAC bearing mice. 3H-Thymidine and 3H-Uridine were employed to study its inhibitory effect on DNA and RNA synthesis respectively in S-180 tumor cells in vitro. RESULTS Alkylating activity of fluorenmustine exceeded that of N-di(2-chloroethyl)amine used as a standard alkylating compound. It has displayed an excellent and reproducible antitumor activity in vivo against EAC and S-180 comparable to that of 5-fluorouracil judging by the increase in median survival times of treated animals. It also significantly increased the life span of mice bearing advanced tumors for 6 days before the drug challenge. However in vitro screening in MCF-7 did not reveal any significant cytotoxicity. The compound did not adversely affect hematopoiesis at its optimum dose. Drug-induced hepatotoxicity and nephrotoxicity were also not detected. It inhibited the synthesis of DNA and RNA in S-180 tumor cells at 8 microM concentration. CONCLUSION Results indicated promising chemotherapeutic potential of Fluorenhymustine.
AIM:To study the humoral immune response to tumor-associated carbohydrate epitopes (TF, Tn and alphaGal) in patients with breast cancer and healthy donors, the putative impact of the chemotherapy and to evaluate if the level of antibody to these epitopes might be beneficial or detrimental for the patients with breast cancer.MATERIALS AND METHODS:The humoral immune response to TF, Tn and alphaGal was studied in 133 patients with breast cancer, including the patients at stage II-III (n = 44) before and after neoadjuvant chemotherapy (10 patients received cyclophosphamide/methotrexate/fluorouracyl (CMF) chemotherapy regimens, 34 patients received cyclophosphamide/doxorubicin/fluorouracil (CAF)), and in controls (healthy donors and patients with fibroadenoma). Fully synthetic carbohydrate hapten-polyacrylamide conjugates were used as antigens in ELISA for anti-carbohydrate antibody determination. The correlation analysis between the level of anti-carbohydrate antibodies and the stage of cancer, histological grade, expression of TF and alphaGal epitopes in tumor tissue, patient's survival was performed.RESULTS:The level of anti-carbohydrate antibodies varied between individuals with no significant correlation between IgG immune response to the three epitopes. Lower levels of antibodies were observed at advanced stages of cancer. Neoadjuvant chemotherapy stimulated antibody production to Tn and alphaGal epitopes (increase > 50%) in about one third of patients. Immunosuppression, decrease in antibody levels, was observed only in 4.5-13.6% of cases. High levels of TF-antigen specific IgG antibody before surgery were associated with a better survival time of stage II breast cancer patients.CONCLUSION:The widely used regimens of neoadjuvant chemotherapy (such as CMF, CAF) can stimulate the immune response to tumor-associated carbohydrate epitopes in some patients. The high levels of anti-TF antibody before surgery are associated with a better survival of stage II breast cancer patients. This may indicate that the selection of immunopotentiating regimens of neoadjuvant chemotherapy might be beneficial for the host.
UNLABELLED:Pathways controling cell proliferation and cell survival require flexible adaptation to environmental stress. Our previous studies showed that latent membrane protein1 (LMP1) encoded by Epstein-Barr virus (EBV) could trigger the expression of Survivin, an apoptosis inhibitor and essential regulator of mitosis. The aim of the work was to analyze the role of Survivin signal pathway in mediating effects triggered by LMP1.METHODS:Tet-on LMP1 HNE2, a tetracycline-regulated LMP1-expression nasopharyngeal carcinoma cell line, was used as cell model. The subcellular location of Survivin was detected by indirect immunofluorescence and Western-blotting assay. Using Ab-knock-out and gene transfection, we introduced anti-sense PS-ODN-Survivin and anti-body to Survivin into the Tet-on LMP1 HNE2, and then the apoptosis and the proliferation of cells were analyzed by flow cytometry, cell colony formation and detection of caspase-3. The results show that upon induction of LMP1 expression, the expression of Survivin in nucleus, level of phosphorylated retinoblastoma gene (Rb), the number of cells in S stage of cell cycle, and the cell colony formation rate were higher than those without LMP1 induction; if the expression of Survivin and the nucleus translocation of Survivin were knocked by introduction of anti-sense PS-ODN-Survivin and anti-Survivin-antibodies respectively, apoptosis rates and the activity of caspase-3 increased.CONCLUSION:LMP1 could trigger the nucleus translocation of Survivin, which led to the shift of S stage and cell proliferation. LMP1 may promote cell proliferation and inhibits apoptosis via Survivin signal pathway.
AIM:Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements are excellent patient-specific targets for clonality studies and monitoring of acute lymphoblastic leukemia (ALL). THE AIMS of the study were to select the optimum panel of primers, evaluate incidence of particular types of monoclonal and oligoclonal gene rearrangements and observe alteration of rearrangement profile between diagnosis of ALL and subsequent relapse(s).METHODS:We used polymerase chain reaction (PCR) for amplification of junctional region of rearranged IgH, TCRD and TCRG genes in combination with heteroduplex analysis in polyacrylamide gel.RESULTS:TCRD gene rearrangements were detected in 64%, TCRG - in 45%, and IgH - in 79% of B-precursor ALL patients. For patients with T-ALL, TCRD gene rearrangements were found in 47%, TCRG gene - in 66%, and IgH - in 19% of cases. Evaluation of biallelic and oligoclonal rearrangements was performed in the study. The highest incidence of oligoclonal rearrangements - 25% was shown for IgH gene in patients with B-precursor ALL. Seven pair cases of patients with de novo leukemia and relapses were analyzed and revealed subclonal deviation in rearrangements of IgH or TCR genes during disease evolution.CONCLUSION:We propose a panel of 13 types of rearrangements (primer pairs) sufficient for tumor cell clonality detection in 96% of patients with ALL. Applications of PCR-based analysis of rearranged IgH, TCRD and TCRG genes for discrimination of mono- and oligoclonality and identification of the origin of relapse were demonstrated.
AIM 1) To evaluate the effect of prostaglandin E2 (PGE2) on the regulation of vascular endothelial growth factor (VEGF) expression in gastric MKN28 cells, and 2) to investigate the role of the epidermal growth factor receptor (EGFR) signal transduction pathway in any effect exerted by PGE2 on VEGF expression. METHODS MKN28 cells were incubated with the vehicle (control) or with PGE2 in the presence or absence of AG1478, a selective inhibitor of EGFR tyrosine kinase, or PD098059, a selective inhibitor of the kinase responsible for ERK2 phosphorylation (mitogen-activated protein (MAP)/extracellular signal-regulated kinase (ERK)). Real-time quantitative polymerase chain reaction and Western blot analysis were used to evaluate VEGF mRNA and protein expression. The activity of EGFR and ERK2 was measured by Western blot analysis. RESULTS PGE2 significantly up-regulated VEGF mRNA and protein expression and increased the activation of EGFR and ERK2. Incubation of MKN28 cells with AG1478 significantly reduced PGE2-induced EGFR activity, ERK2 activity, and VEGF mRNA and protein expression. Meanwhile, incubation of MKN28 with PD098059 reduced PGE2-induced ERK2 activity and VEGF mRNA and protein expression, but had no effect on EGFR activity. CONCLUSION Our data suggested that PGE2 up-regulates VEGF expression in gastric cancer cells via transactivation of EGFR-MAPK signaling pathways, which may be mechanisms underlying the contribution of COX-2 to tumor angiogenesis in gastric cancer.
Aim: To identify novel tuberous sclerosis complex (TSC2) binding partners by yeast two-hybrid screening. Methods: The yeast two-hybrid system DupLEX-A (TM) developed by OriGene Technologies and Mouse embryo and HeLa cells cDNA libraries were used in this study. The "bait" constructs, containing full-length and truncated form of TSC2 were prepared. The expression of all constructs in yeast was confirmed by immunoblotting with specific anti-LexA antibodies. The suitability of generated constructs for screening was tested in autoactivation and nuclear translocation assays. Screening of mouse embryo and HeLa cDNA libraries with selected baits was carried out according to manufacturer's recommendations. Positive clones were selected using double selection procedure and further confirmed in mating assay. Isolated cDNA clones were identified by automated DNA sequencing and database searching. Results: Extensive screening of two cDNA libraries from mouse embryo and HeLa cells with TSC2 baits led to the isolation of 102 positives clones. The specificity of interaction between TSC2 and binding proteins of selected clones was confirmed by mating assay for 83 clones. Sequencing of these clones indicated that they encode already known and novel TSC2-binding partners. Conclusion: The isolation of several known TSC2-binding partners, such as several isoforms of 14-3-3, demonstrates the validity of generated bait constructs and screening conditions. In addition, we have found a number of novel interactors, which encode cytoskeletal proteins and signaling molecules, such as Ser/Thr phosphatases.
Background. Measurements of DNA double strand breaks and their subsequent repair after in vitro irradiation has been suggested to be an alternative way of monitoring radiotherapeutic response. Methods: In the present study, the DNA repair kinetics (using a neutral version of the Comet assay) up to 45 min after a single dose of 2 Gy was studied as well as the gene expression profiles, before and 45 min after the irradiation, in two human lung cancer cell lines with different radiosensitivity (U-1285 and U-1810). Results: Immediately after the irradiation, both cell lines responded with increased levels of DNA damage. However, the induced damage was slightly higher in U4810 (known to be radioresistant) than in U-1285 (known to be radiosensitive), and the latter cell line also seemed to have a slightly more efficient DNA-repair. The two different lung cancer cell lines were highly heterogeneous in gene expression, both before and after the irradiation, and there was no obvious relationship between the Comet data and the microarray data. Conclusion: Given the fact that U-1810 has been classified as radioresistant and U-1285 as radiosensitive in clonogenic assays, the results of the present study indicate that radiation-induced DNA double strand breaks and DNA-repair efficiency are poor indicators of the intrinsic radiosensitivity of human lung cancer cells irradiated with a single dose in vitro.
BACKGROUND AND AIM Fusion of human dendritic cells (DCs) with tumor cells is an effective approach for delivering tumor antigens to DCs, and DC/tumor fusion cells are potent stimulators of autologous T cells. However, the integration and morphology of DC/osteosarcoma fusion cells has not been examined. This study was designed to investigate the antitumor effects of tumor vaccine produced by electrofusion between human osteosarcoma cells and DCs. METHODS In the present study, we eletrofused patient-derived DCs to autologous osteosarcoma cells. The fusion cells possessed the properties of both patient cells. After electrofusion, the cytoplasm of the two cells was integrated, whereas their nuclei remained separate entities. The intracellular structure was observed on fusion cells under the transmission electron microscope. RESULTS Coculture of patient-derived peripheral blood mononuclear cells (PBMC) with DC/tumor fusion cells resulted in activation of T cells as assessed by standard cytotoxic T lymphocytes (CTLs) assays. CONCLUSIONS The present study provides valid evidence on integration of human DCs and tumor cells and links their properties to T cell activation. The fusion cells may thus represent a promising strategy for DC-based immunotherapy of patients with osteosarcoma.
