Tyrosine Kinase Inhibitors (TKIs) in Lung Cancer Treatment: A Comprehensive Analysis.

Lung cancer is the leading type of cancer worldwide today. Kinases play a crucial role in mediating the signaling pathways, and it directs to control several necessary cellular processes. Conversely, the deregulation of tyrosine kinases leads to oncogenic conversion, uncontrolled cell proliferation and tumorigenesis. Tyrosine kinases are largely deregulated in lung cancer and specifically in non-small cell lung cancer (NSCLC). Therefore, the inhibition of pathogenic kinases is a breakthrough development in cancer research, treatment and care, which clinically improve the quality of life. In the last decades, various single or combination inhibitors are approved by U.S Food and Drug Administration (FDA) and commercially available in clinics, and currently, several preclinical studies are ongoing and examining the kinase inhibitors. However, many gaps remain in understanding the mechanisms of kinase inhibitors and their selectivity. In this analysis, we focus on a class of receptor and non-receptor tyrosine kinase inhibitors and their novel role in lung cancer.

Anti-diabetic activity of crude polysaccharide and rhamnose-enriched polysaccharide from G. lithophila on Streptozotocin (STZ)-induced in Wistar rats.

The aim of the present study was to elucidate the anti-diabetic effects of the crude polysaccharide and rhamnose-enriched polysaccharide derived from G. lithophila on streptozotocin (STZ)-induced diabetic Wistar rats. Treatment with crude polysaccharide and rhamnose-enriched polysaccharide showed increases in body weight and pancreatic insulin levels and a decrease in blood glucose levels compared with control diabetic rats. The blood concentrations of total cholesterol (TC), triglycerides (TGs), low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) decreased, and high-density lipoprotein (HDL) increased both in the crude polysaccharide- and rhamnose-enriched polysaccharide-treated rats. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels increased, and malondialdehyde (MDA) levels decreased in the livers, kidneys and pancreases of crude polysaccharide- and rhamnose-enriched polysaccharide-treated rats. Immunohistological examination further confirmed that restoration of the normal cellular size of the islets of Langerhans and the rebirth of ß-cells were found to be greater in the body region than in the head and tail regions of the pancreas. The crude polysaccharide- and rhamnose-enriched polysaccharide-treated diabetic rats showed normal blood glucose levels and insulin production, and reversed cholesterol levels and enzymatic actions. Therefore, rhamnose-enriched polysaccharide from G. lithophila acts as a potent anti-diabetic agent to treat diabetes and can lead to the development of an alternative medicine for diabetes in the future.

Meta-analysis of functional expression and mutational analysis of c-Met in various cancers.

Comprehensive genomic profiling is expected to revolutionize cancer therapy. c-Met signaling is responsible for tumorigenesis in various cancers. In this prospective, we present the prevalence of c-Met mutations and copy number alterations across various solid tumors. We used major databases like cBioportal, PubMed, and COSMIC for c-Met mutation and amplification data collection from various cancers. Our result shows complete details about c-Met mutation and its clinical data of various cancers. Hotspot mutation of human c-Met protein reveals that repeatedly and most mutated regions and these hotspots may be a diagnostic tool for cancer confirmation. Amino acid and nucleotide changes and their prevalence were reported in a number of individual cancers. However, we collectively present the amino acid and nucleotide changes in various cancers in this review. Our collection of data for c-Met mutation and its distribution in different cancer tissue is showing that the missense mutation is the major one in all type of cancers. Copy number variation data showing amplification and deletion of human c-Met from various tumor types, lung and central nervous system tumors showing high amplification comparatively other types.

Multi-elemental concentration in different body parts of Sepiella inermis by inductively coupled plasma mass spectrometry.

The present study examined the accumulation of metal on Sepiella inermis from the Mudasalodai Landing Center, from southeast coastal region of India. Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine metals including aluminium, boron, cadmium, cobalt, chromium, copper, iron, manganese, magnesium, nickel, lead and zinc. The results showed that metal concentration in S. inermis detected in the head, arm, mantle, eye, ink, liver and nidamental gland with higher concentration of magnesium up to 992.78 mg/kg, and tentacle showed maximum concentration of aluminium 306.72 mg/kg. Further, copper found in low concentration ranges from 0.04 to 0.55 mg/kg in different parts of S. inermis. Heavy metal like cadmium detected high in tentacle with 0.24 mg/kg, and the manganese present in eye was 0.55 mg/kg. However, no accumulation of nickel was found in the tentacle part.

Nanoparticles from Actinobacteria: A Potential Target to Antimicrobial Therapy.

Nanoparticles have gained significant importance in the past two decades, due to their multifaceted applications in the field of nanomedicine. As our ecosystems and habitats are changing due to global warming, many new diseases are emerging continuously. Treating these costs a lot of money and mostly ends up in failure. In addition, frequent use of antibiotics to control the emerging diseases has led the pathogens to develop resistance to antibiotics. Hence, the nanoparticles are targeted to treat such diseases instead of the costly antibiotics. In particular, the biosynthesized nanoparticles have received considerable attention due to their simple, eco-friendly and promising activity. To highlight, microbial mediated nanoparticles have been found to possess higher activity and thus have a promising role in antimicrobial therapy to fight against the emerging drug-resistant pathogens. In this context, this review article is aimed at highlight the role of nanoparticles in the field of nanomedicine and importance of actinobacteria in the nanoparticle synthesis and their need in antimicrobial therapy. This is a comprehensive review, focusing on the potential of actinobacteria-mediated nanoparticles in the field of nanomedicine.

Bioremediation of wastewater through a quorum sensing triggered MFC: A sustainable measure for waste to energy concept.

A mission for fast advancement has constrained us to unpredictably tap various natural assets. The reckless utilisation of fossil fuels led unmanageable wastes which have greatly affected our health and environment. Endeavours to address these difficulties have conveyed to the frontal area certain creative natural solutions particularly the utilisation of microbial digestion systems. In the previous two decades, the microbial fuel cell (MFC) innovation has caught the consideration of the researchers. The MFCs is a kind of bio-electrochemical framework with novel highlights, for example, power production, wastewater treatment, and biosensor applications. Lately, dynamic patterns in MFC inquire about on its synthetic, electrochemical, and microbiological perspectives have brought about its observable applications. The MFCs have begun as a logical interest, and in numerous regards, these remaining parts to be the situation. This is especially a result of the multidimensional uses of this eco-accommodating innovation. The innovation relies upon the electroactive microorganisms, prominently known as exoelectrogens. In the first place, it is the main innovation that can create energy out of waste, without the contribution of outer/extra energy. Modification of electrodes with nanomaterials, for example, gold nanoparticles and iron oxide nanoparticles or pretreatment techniques, for example, sonication and autoclave disinfection have indicated promising outcomes in improving MFC execution for power generation and wastewater treatment. The MFC innovation has been likewise explored for the remediation of different heavy metals and hazardous components, and to recognize the poisonous components in wastewater. What's more, the MFCs can be adjusted into microbial electrolysis cells to produce hydrogen energy from different natural sources. This article gives a thorough and cutting-edge appraisal of the novel magnitudes of the MFC.

Chemical structure and biological properties of a polysaccharide isolated from Pleurotus sajor-caju.

Herein, a polysaccharide obtained from Pleurotus sajor-caju was fractionated via anion-exchange column chromatography and purified using gel permeation column chromatography. The chemical characterization of the polysaccharide indicated that it contained 90.16% total carbohydrate, 0% protein, 12.7% ash and 5.2% moisture; on the other hand, the carbon, hydrogen and nitrogen contents were found to be 31.53, 4.28 and 3.01%, respectively. The polysaccharide has the molecular weight of 79 kDa; the chemical structure of the polysaccharide is →6)α-d-Glciv(1→6)α-d-Glciii(1→6)ß-d-Glcii(1→6)α-d-Glci(1→units. The polysaccharide exhibited the DPPH radical scavenging activity of 21.67-68.35% at 10-160 µg ml-1, ABTS radical scavenging activity of 16.01-70.09% at 25-125 µg ml-1, superoxide radical scavenging activity of 24.31-73.64% at 50-250 µg ml-1, hydroxyl radical scavenging activity of 16.64-63.51% at 25-125 µg ml-1 and reducing power of 0.366-1.678% at 10-120 µg ml; further evaluation of the polysaccharide revealed its anticancer activity of 18.61-63.21% at 100-500 µg ml-1 concentration against the AGS human gastric carcinoma cell line. The active principle of the polysaccharide may be used in the food and pharmacological industry in the future.

Inhibition of EGFR-AKT axis results in the suppression of ovarian tumors in vitro and in preclinical mouse model.

Ovarian cancer is the leading cause of cancer related deaths in women. Genetic alterations including overexpression of EGFR play a crucial role in ovarian carcinogenesis. Here we evaluated the effect of phenethyl isothiocyanate (PEITC) in ovarian tumor cells in vitro and in vivo. Oral administration of 12 µmol PEITC resulted in drastically suppressing ovarian tumor growth in a preclinical mouse model. Our in vitro studies demonstrated that PEITC suppress the growth of SKOV-3, OVCAR-3 and TOV-21G human ovarian cancer cells by inducing apoptosis in a concentration-dependent manner. Growth inhibitory effects of PEITC were mediated by inhibition of EGFR and AKT, which are known to be overexpressed in ovarian tumors. PEITC treatment caused significant down regulation of constitutive protein levels as well as phosphorylation of EGFR at Tyr1068 in various ovarian cancer cells. In addition, PEITC treatment drastically reduced the phosphorylation of AKT which is downstream to EGFR and disrupted mTOR signaling. PEITC treatment also inhibited the kinase activity of AKT as observed by the down regulation of p-GSK in OVCAR-3 and TOV-21G cells. AKT overexpression or TGF treatment blocked PEITC induced apoptosis in ovarian cancer cells. These results suggest that PEITC targets EGFR/AKT pathway in our model. In conclusion, our study suggests that PEITC could be used alone or in combination with other therapeutic agents to treat ovarian cancer.

Role of protein kinase C ß and vascular endothelial growth factor receptor in malignant pleural mesothelioma: Therapeutic implications and the usefulness of Caenorhabditis elegans model organism.

PURPOSE: To examine the role of both protein kinase C (PKC)-ß and vascular endothelial growth factor receptor (VEGFR)-2 in malignant pleural mesothelioma (MPM) using respective inhibitors, enzastaurin and KRN633. MATERIALS AND METHODS: MPM cell lines, control cells, and a variety of archived MPM tumor samples were used to determine the protein expression levels of PKC-ß, VEGFR-2, VEGF, and p-AKT. Effects of enzastaurin and KRN633 on phosphorylation status of key signaling molecules and viability of the mesothelioma cells were determined. The common soil nematode, Caenorhabditis elegans, was treated with enzastaurin to determine its suitability to screen for highly potent kinase inhibitors. RESULTS: PKC-ß1, PKC-ß2 and VEGFR-2/KDR were overexpressed in MPM cell lines and MPM tumor tissues. Enzastaurin treatment resulted in significant loss in viability of VEGF induced cell proliferation; however, the effect of KRN633 was much less. Enzastaurin also dramatically decreased the phosphorylation of PKC-ß, its downstream target p-AKT, and surprisingly, the upstream VEGFR-2. The combination of the two drugs at best was additive and similar results were obtained with respect to cell viability. Treatment of C. elegans with enzastaurin resulted in clear phenotypic changes and the worms were hypermotile with abnormal pattern and shape of eggs, suggesting altered fecundity. CONCLUSIONS: PKC-ß1 and VEGFR-2 are both excellent therapeutic targets in MPM. Enzastaurin was better at killing MPM cells than KRN633 and the combination lacked synergy. In addition, we show here that C. elegans can be used to screen for the next generation inhibitors as treatment with enzastaurin resulted in clear phenotypic changes that could be assayed.

Ethnic differences and functional analysis of MET mutations in lung cancer.

PURPOSE: African Americans have higher incidence and poorer response to lung cancer treatment compared with Caucasians. However, the underlying molecular mechanisms for the significant ethnic difference are not known. The present study examines the ethnic differences in the type and frequency of MET proto-oncogene (MET) mutation in lung cancer and correlated them with other frequently mutated genes such as epidermal growth factor receptor (EGFR), KRAS2, and TP53. EXPERIMENTAL DESIGN: Using tumor tissue genomic DNA from 141 Asian, 76 Caucasian, and 66 African American lung cancer patients, exons coding for MET and EGFR were PCR amplified, and mutations were detected by sequencing. Mutation carriers were further screened for KRAS2 and TP53 mutations. Functional implications of important MET mutations were explored by molecular modeling and hepatocyte growth factor binding studies. RESULTS: Unlike the frequently encountered somatic mutations in EGFR, MET mutations in lung tumors were germline. MET-N375S, the most frequent mutation of MET, occurred in 13% of East Asians compared with none in African Americans. The frequency of MET mutations was highest among male smokers and squamous cell carcinoma. The MET-N375S mutation seems to confer resistance to MET inhibition based on hepatocyte growth factor ligand binding, molecular modeling, and apoptotic susceptibility to MET inhibitor studies. CONCLUSIONS: MET in lung cancer tissues contained nonsynonymous mutations in the semaphorin and juxtamembrane domains but not in the tyrosine kinase domain. All the MET mutations were germline. East Asians, African-Americans, and Caucasians had different MET genotypes and haplotypes. MET mutations in the semaphorin domain affected ligand binding.

PAX5 is expressed in small-cell lung cancer and positively regulates c-Met transcription.

PAX5 is a nuclear transcription factor required for B cell development, and its expression was evaluated in upper aerodigestive malignancies and pancreatic cancer by immunoblotting. The PAX5 protein expression was relatively strong in small-cell lung cancer (SCLC, 11/12); however, its expression was not detected in non-SCLC (NSCLC, n=13), mesothelioma (n=7), pancreatic (n=6), esophageal (n=6) and head and neck cancer cell lines (n=12). In comparison, PAX8 and PAX3 expressions were absent or non-detectable in SCLC cell lines; however, PAX8 was expressed in most of the tested NSCLC cell lines (13/13) and also frequently in all the other cell lines. We also detected frequent expressions of PAX2 and PAX9 protein in the various cell lines. Utilizing neuroendocrine tumor samples, we found that the frequency as well as the average intensity of the expression of PAX5 increased from pulmonary carcinoid (9%, moderate and strong PAX5 expression, n=44), to large-cell neuroendocrine carcinoma (LCNC, 27%, n=11) to SCLC (33%, n=76). FISH analysis revealed no translocations of the PAX5 gene, but polyploidy in some SCLC tumor tissues (6/37). We determined that PAX5 could regulate the transcription of c-Met using luciferase-coupled reporter and chromatin immunoprecipitation analysis. In addition, the phospho-c-Met (active form) and PAX5 were both localized to the same intra-nuclear compartment in hepatocyte growth factor treated SCLC cells and interacted with each other. Finally, we determined the therapeutic translational potential of PAX5 using PAX5 knockdown SCLC cells in conjunction with Topoisomerase 1 (SN38) and c-Met (SU11274) inhibitors. Loss of endogenous PAX5 significantly decreased the viability of SCLC cells, especially when combined with SN38 or SU11274, and maximum effect was seen when both inhibitors were used. Therefore, we propose that PAX5 could be an important regulator of c-Met transcription and a potential target for therapy in SCLC.

Protein kinase C beta in malignant pleural mesothelioma.

Malignant pleural mesothelioma (MPM) is a disease with few therapeutic options. Protein kinase C beta (PKCbeta) is involved in important cellular functions. Enzastaurin (LY317615.HCl) is a novel inhibitor of PKC in clinical development. MPM cell lines (7) and patient tumor tissues (24) were evaluated for expression of PKCbeta by immunoblotting and immunohistochemistry, respectively. In-vitro cell growth assays were performed with enzastaurin with or without cisplatin. Cell migration was evaluated with the wound healing assay. Downstream signaling (survival and focal adhesion pathways) was studied by immunoblotting for related molecules in the presence of phorbol ester with or without enzastaurin. Expression for PKCbeta1 was seen in all cases, with a mean integrated optical density of 152.5 (standard deviation=95.47, n=24), whereas PKCbeta2 expression was less intense, with a mean integrated optical density of 11.45 (standard deviation=16.27, n=21). There was a trend toward lower overall survival among patients expressing above-median PKCbeta1 (P=0.064), but not PKCbeta2. Robust expression of PKCbeta1 and low expression of PKCbeta2 were observed in MPM cell lines. Treatment of MPM cell lines with enzastaurin revealed an IC50 of 5 micromol/l, and strong synergism was observed when combined with cisplatin. Wound healing assay revealed that treatment of H2461 cells with enzastaurin reduced migration by 59.2%. Enzastaurin treatment led to disruption of F-actin architecture. Downstream signaling showed reduced phosphorylation of AKT, FAK (focal adhesion kinase), p130Cas, S6 ribosomal protein, and paxillin. PKCbeta1 was expressed in the majority of MPM samples. Enzastaurin has preclinical activity against MPM, and exhibited synergism with cisplatin. PKCbeta inhibition in MPM might be able to reduce the invasiveness of MPM by affecting cytoskeletal function.

C. elegans as a model organism for in vivo screening in cancer: effects of human c-Met in lung cancer affect C. elegans vulva phenotypes.

Cancers typically harbour several mutant forms of key cellular genes that contribute to its complex phenotype. Our lab has previously identified gain-of-function mutations in some of the receptor tyrosine kinases such as c-Met in lung cancer. In order to investigate the mutant gene in the context of a whole organism, the current choice of in vivo model is limited to the mouse. To rapidly screen the functional aspects of mutant forms of c-Met detected in lung cancer, we used the nematode C. elegans as the model organism. Transgenic worms were generated that harbour wild type or the frequently seen mutant forms of c-Met in lung cancer (c-MetR988C and c-MetT1010I). Expression of the mutant human c-Met forms in C. elegans consistently resulted in significantly low fecundity and abnormal vulval development characterized by hyperplasia. Interestingly, exposure of c-Met mutant transgenic worms to nicotine resulted in enhanced abnormal vulval development, fecundity and locomotion. Our studies provide first evidence that human c-Met mutations can be studied in C. elegans, and that carcinogens can enhance mutant c-Met function expressed in C. elegans transgenic animals. We therefore propose the use of C. elegans as a model to rapidly assess the role of cancer specific gene mutations in the context of a whole organism.

Paxillin is a target for somatic mutations in lung cancer: implications for cell growth and invasion.

Lung cancer is characterized by abnormal cell growth and invasion, and the actin cytoskeleton plays a major role in these processes. The focal adhesion protein paxillin is a target of a number of oncogenes involved in key signal transduction and important in cell motility and migration. In lung cancer tissues, we have found that paxillin was highly expressed (compared with normal lung), amplified (12.1%, 8 of 66) and correlated with increased MET and epidermal growth factor receptor (EGFR) gene copy numbers, or mutated (somatic mutation rate of 9.4%, 18 of 191). Paxillin mutations (19 of 21) were clustered between LD motifs 1 and 2 and the LIM domains. The most frequent point mutation (A127T) enhanced lung cancer cell growth, colony formation, focal adhesion formation, and colocalized with Bcl-2 in vitro. Gene silencing from RNA interference of mutant paxillin led to reduction of cell viability. A murine in vivo xenograft model of A127T paxillin showed an increase in tumor growth, cell proliferation, and invasion. These results establish an important role for paxillin in lung cancer.

Benzyl isothiocyanate-induced DNA damage causes G2/M cell cycle arrest and apoptosis in human pancreatic cancer cells.

Benzyl isothiocyanate (BITC) has been shown to inhibit chemically induced pancreatic cancer in experimental animals. However, the mechanism responsible for the anticancer effects of BITC is not clearly understood. In this study, we tested whether BITC treatment would affect the growth of Capan-2 human pancreatic cancer cells. BITC (10 micromol/L) treatment caused marked phosphorylation of H2A.x (2.6-fold) and permanent damage to Capan-2 cells. BITC-mediated G2/M arrest was associated with up-regulation of cyclin dependent kinase inhibitor p21(Waf1/Cip1) and the activation of checkpoint kinase 2, whereas the expressions of other G2/M regulatory proteins, including CyclinB1, Cdc2, and cell division cycle 25C (Cdc25C), were down-regulated by 19, 51, and 70%, respectively, compared with control. These changes resulted in a 55% inhibition of Cdc2 kinase activity. In addition, the decline in the expression of Cdc25C was completely blocked when the cells were treated with lactacystin (proteasome inhibitor) prior to BITC treatment. However, G2/M arrest and apoptosis induced by BITC were partially blocked by pretreatment of cells with lactacystin. Taken together, the results of this study suggest the involvement of multiple signaling pathways targeted by BITC in mediating G2/M cell cycle arrest and apoptosis in Capan-2 cells and warrant further investigation.