Search

Hyaluronic acid-based nanoparticles can be promising tools for gene delivery in in vivo reprogramming CAR T cells, as well-designed carriers are needed for targeting and transducing circulating T cells.

Cisplatin (CDDP) induces senescence characterized by senescence-associated secretory phenotypes (SASP) and the unfolded protein response (UPR). In this study, we investigated the proteins related to the UPR during the senescence cell fate. Strikingly, we found that one of the critical ER-resident proteins, GRP78/BiP, was significantly altered. Here we show that GRP78 levels differentially expressed depending on non-small lung cancer subtypes. GRP78 indeed regulates the evasion of senescence in adenocarcinoma A549 cells, in which the increased GRP78 levels enable them to re-proliferate after CDDP removal. Conversely, GRP78 is downregulated in the senescence H460 cells, making them lacking senescence evasion capability. We observed that the translational regulation critically contributed to the GRP78 protein levels in CDDP-induces senescence. Furthermore, the increased GRP78 level during senescence confers resistance to senolytic drug, Bortezomib, as observed by a twofold increase in IC50 in A549 senescence cells compared to the wild-type. This observation is also consistent in the cells that have undergone genetic manipulation by transfection with pcDNA3.1(+)-GRP78/BiP plasmids and pSpCas9(BB)-2A-Puro containing guide RNA sequence targeting GRP78 exon 3 to induce the overexpression and downregulation of GRP78 in H460 cells, respectively. Our findings reveal a unique role of GRP78 on the senescence evasion cell fate and senolytic drug resistance after cisplatin-based chemotherapy.

During an investigation of rare actinobacteria, isolate SC076T was isolated from a soil sample collected from Sichang Island, Chonburi Province, Thailand. The strain showed the highest 16S rRNA gene similarity to Saccharothrix australiensis DSM 43800T (98.6%) and Saccharothrix espanaensis DSM 44229T (98.6%). The zigzag morphology of the spore chain was observed on the aerial mycelia. meso-Diaminopimelic acid was detected in the peptidoglycan. Whole-cell sugars contained rhamnose, ribose, mannose glucose and galactose. Polar lipids were phosphatidylethanolamine, hydroxyphosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol mannoside, phosphatidylinositol, unidentified ninhydrin-positive glycolipid, unidentified glycolipid and four unidentified lipids. The menaquinones were MK-9(H8), MK-9(H4), MK-9(H2) and MK-9(H0). The predominant fatty acids were iso-C16 : 0, iso-C15 : 0 and anteiso-C17 : 0. The draft genome of SC076T was 8040245 bp with a G+C content of 72.5 mol%. The results of genomic analysis between strain SC076T and the related type strains showed that the digital DNA–DNA hybridization and average nucleotide identity values among the strains were 23.6-32.8% and 77.7–86.8 %, respectively, which are lower than the thresholds used to distinguish strains from others of the same species. Based on the taxonomic evidence, strain SC076T represents a novel species of the genus Saccharothrix for which the name Saccharothrix obliqua sp. nov. is proposed. The type strain is SC076T (=TBRC 14540T=NBRC 115117T).

Heart failure (HF) has reached epidemic proportions in developed countries, affecting over 20 million people worldwide. Despite modern medical and device therapies, 60–70% of HF patients still die within 5 years of diagnosis as it relentlessly progresses through pervasive apoptotic loss of cardiomyocytes. Although fortilin, a 172-amino-acid anti-p53 molecule, is one of the most expressed proteins in the heart, its precise role there has remained unknown. Also unclear is how cardiomyocytes are protected against apoptosis. Here, we report that failing human hearts express less fortilin than do non-failing hearts. We also found that mice lacking fortilin in the heart (fortilinKO-heart) die by 9 weeks of age due to extensive cardiomyocyte apoptosis and severe HF, which suggests that fortilin sustains cardiomyocyte viability. The lack of fortilin is also associated with drastic upregulation of p53 target genes in the hearts. The heart-specific deletion of p53 in fortilinKO-heart mice extends their life spans from 9 to 18 weeks by mitigating cardiomyocyte apoptosis. Our data suggest that fortilin is a novel cardiac p53 inhibitor and that its inadequate expression in failing hearts and subsequent overactivation of the p53 apoptosis pathway in cardiomyocytes exacerbates HF.

The endoplasmic reticulum, the cytoplasmic organelle that matures a massive amount of nascent secretory polypeptides, is particularly sensitive to stress. Endoplasmic reticulum stress causes unfolded proteins to populate the organelle, eliciting the unfolded protein response. During the unfolded protein response, GRP78—an endoplasmic reticulum master stress regulator—detaches from three endoplasmic reticulum stress sensors (IRE1α, PERK, and ATF6) and allows them to activate the apoptotic signaling pathway. Fortilin, a pro-survival molecule, is known to inhibit apoptosis by binding and inhibiting p53, but its role in endoplasmic reticulum stress-induced apoptosis remains unknown. Here, we report that fortilin directly interacts with the cytoplasmic domain of IRE1α, inhibits both kinase and endoribonuclease (RNase) activities of the stress sensor, and protects cells against apoptotic cell death at both cellular and whole animal levels. Our data support a role of fortilin in the unfolded protein response and its potential participation in human diseases caused by unfolded protein response., IRE1α is an ER stress sensor, whose activity induces apoptosis. Here, the authors report that fortilin, a pro-survival factor, with yet unknown roles in ER stress, interacts with active IRE1α, inhibits both its kinase end RNase activities, and protects cells from apoptosis both in vitro and in vivo.

Lung cancer is one of leading cancer-related causes of mortality worldwide. The propensity for metastasis in this disease undoubtedly contributes to the low survival rate and final poor outcome of patients. Lung cancer metastasis involves specific molecular pathways that confer the cancer cells abilities to survive in circulation, be motile, and form new tumors. Recent evidences have widened the understanding of cancer disseminating basis and emphasized the role of genetic and epigenetic imperfections, epithelial to mesenchymal transition, and cancer stem cells. The present chapter aims to illustrate metastatic processes of lung cancer, shedding more light on such key molecular paradigms. A deeper understanding of the mechanisms of lung cancer metastasis may lead to novel strategies in cancer preventive and therapeutic approaches.

Epithelial-mesenchymal transition (EMT) is implicated in cancer pathological processes, particularly cancer invasion and metastasis. The present study demonstrated that EMT was critical for the metastasic potential of lung cancer cells isolated from a patient. P1 primary lung cancer cells were found to exhibit increased anoikis resistance compared with established A549, H23 and H460 lung cancer cells. Results of migration and invasion assays revealed that the invasion capability of P1 and A549 cells was higher than that of H23 and H460 cells. However, the migration of P1 cells was similar to that of H23 and H460 cells while A549 demonstrated a superior migrating ability. Western blot analysis indicated that while E-cadherin levels in all lung cancer cells were identified as comparable, P1 cells expressed the highest levels of N-cadherin. In the present study, detachment of cells was demonstrated for the first time to stimulate further transition of E-cadherin to N-cadherin. In addition, this obervation was more pronounced in P1 cells. These observations highlight the importance of EMT in cancer metastasis. In order to study the effect of ethnicity on cancer cell behavior, in the future a large number of Thai patient-derived cell lines must be analyzed.

169 ISSN 1758-1966 10.2217/LMT.13.13 © 2013 Future Medicine Ltd Lung Cancer Manage. (2013) 2(3), 169–171 The translocation of cancer cells to other parts of the body is the process of cancer metastasis, and this term has frightened people with lung cancer for years. Recently, any method that prevents cancer metas­ tasis is of great interest in cancer research prospects. Based on the fact that without metastasis, the tumor can be eliminated entirely at its primary site, a better under­ standing of the body’s key defense mecha­ nism against metastasis, named anoikis, may result in new strategies that prevent the progression of cancers, aid conven­ tional cancer treatment and reduce cancer relapse.

Background The enhancement of migration is critical for facilitating cancer cell metastasis. Method Lung cancer H23 cells were transfected with either a caveolin-1 (Cav-1) overexpression or shCav-1 plasmid and further subjected to cell migration assays and lamellipodia characterization. The regulation of Cav-1 via an ATP-dependent tyrosine kinase (Akt) pathway was further examined by Akt knockdown in Cav-1 overexpressing cells and migratory behavior investigations. Results Here, we demonstrate for the first time that overexpression of Cav-1 in human lung cancer H23 cells significantly increased the formation of lamellipodia, whereas the suppression of Cav-1 using shRNA transfection had the opposite effect. Consistent with an increase in lamellipodia, Cav-1 overexpressing cells exhibited increased migratory activity in comparison to their parental, control-transfected, H23 cells. The induction of lamellipodia was demonstrated to occur via the Akt pathway because the addition of the Akt inhibitor LY294002 inhibited lamellipodia in both Cav-1-overexpressing and H23 cells. Additionally, transient transfection with Akt-siRNA significantly inhibited the formation of lamellipodia and the migration of Cav-1-overexpressing H23 cells. In addition, Cav-1 levels and the migratory action of other lung cancer cells, namely, H460 and A549, were assessed, and the migration of these cells was found to be correlated with the basal Cav-1 level. Conclusion These data showed that Cav-1 enhances cancer cell migration through Akt-mediated lamellipodia formation. Our results provide novel insights regarding the molecular mechanism controlling cancer cell migration, leading to a better understanding of cancer cell biology.

Nitric oxide (NO) in tumor microenvironment may have a significant impact on metastatic behaviors of cancer. Noncytotoxic doses of NO enhanced anoikis resistance and migration in lung cancer H23 cells via an increase in lamellipodia, epithelial-mesenchymal transition (EMT) markers including vimentin and snail, and caveolin-1 (Cav-1). However, the induction of EMT was found in Cav-1-knock down cells treated with NO, suggesting that EMT was through Cav-1-independent pathway. These effects of NO were consistently observed in other lung cancer cells including H292 and H460 cells. These findings highlight the novel role of NO on EMT and metastatic behaviors of cancer cells.

The effect of extended exposure of cancer cells to nitric oxide (NO), an endogenous mediator frequently found increased in tumors, is largely unknown. In the present study, the effect of long-term NO exposure on chemotherapeutic resistance was investigated in lung cancer cells.The effect of long-term exposure of human lung cancer cells to NO on susceptibility to chemotherapeutic agents-induced apoptosis was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, co-staining of Hoechst 33342 and propidium iodide (PI), and Annexin V detection. The expression of survival-related proteins was analyzed by western blot. Gene manipulation was used for evaluation of the effect of expression proteins on susceptibility of the cells to chemotherapeutic agent-mediated death.Long-term NO exposure for 7-14 days rendered the lung cancer cells resistant to cisplatin, doxorubicin, and etoposide dose- and time-dependently. The underlying mechanism was found to involve the adaptive responses of the cells, by increasing survival due to increase in the level of caveolin-1 (CAV1) and anti-apoptotic B-cell lymphoma-2 (BCL2), and up-regulation of activated protein kinase B (AKT). The gene manipulation study revealed that the increase of activated AKT and BCL2 was responsible for the resistance to all tested drugs, while the up-regulation of CAV1 only attenuated cell death mediated by doxorubicin and etoposide. Interestingly, NO-mediated drug resistance was found to be reversible when cells were further cultured in the absence of NO for five days.These findings reveal the novel role of NO in the tumor environment, in attenuating chemotherapeutic susceptibility and this could be beneficial in contriving strategies to treat the disease.

Knowledge regarding substances that attenuate motility of cancer cells has gathered significant attention, as they benefit the development of novel anticancer strategies. The anti-migration and anti-invasion activities of artonin E, extracted from bark of Artocarpus gomezianus, were investigated in lung cancer cells in this study.Cytotoxicity and antiproliferative effects of artonin E were examined by 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Migration and invasion assays were performed on H460, H23, A549 and H292 human lung cancer cells. Cell morphology was determined by phalloidin-rhodamine staining. Motility-related proteins were investigated by western blotting.Artonin E exhibited anti-migration and anti-invasion activities in H460 cells. Cell morphology revealed that treatment of the cells with non-toxic concentrations of artonin E resulted in a decrease of activated focal adhesion kinase (FAK), downstream protein kinase B (AKT) activation, and Cell division cycle-42 (CDC42), all of which were associated with the anti-motility effect of this compound. Artonin E inhibited invasion and migration of other lung cancer cells, namely H292, H23 and A549 cells.These results suggest that artonin E may be a promising candidate for anti-metastasis use.

Nitric oxide (NO) found in the vicinity of lung cancer cells may play a role in the regulation of cancer cell behaviors. To explore the possible effects of NO on cell motility, human lung cancer cells were exposed to nontoxic concentrations of NO for 0–14 days, and the migratory characteristics of the cells were determined. The present study found that long-term treatment with NO significantly enhanced cell migration in a dose- and time-dependent manner. Furthermore, we found that the increased migratory action was associated with the increased expression of caveolin-1 (Cav-1), which in turn activated the focal adhesion kinase (FAK) and ATP-dependent tyrosine kinase (Akt) pathways. Notably, the NO-treated cells exhibited an increased number of filopodia per cell, as well as an increase in the levels of cell division cycle 42 (Cdc42) protein. Together, these results indicate that extended NO exposure has a novel effect on cell migration through a Cav-1-dependent mechanism, a finding that strengthens our understanding of cancer biology.

Anoikis, or detachment-induced apoptosis, is recognized as a key inhibitory process of cancer metastasis. Since lung cancer cells possess an ability to resist anoikis, resulting in a high rate of metastasis and death, the present study aimed to investigate the possible anoikis-sensitizing effect of artonin E (AE).AE was extracted from bark of Artocarpus gomezianus. Anoikis sensitization of AE was investigated in H460, A549 and H292 human lung cancer cells. The level of anoikis-related proteins was determined by western blot analysis and viable cells were measured by the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) method.AE was shown to enhance anoikis of H460 cells in a dose-dependent manner. We investigated the underlying mechanisms of AE on anoikis sensitization and found that AE sensitized the cells by down-regulating the anti-apoptotic myeloid leukemia cell sequence-1 (MCL1) protein but had no significant effect on other proteins of the B-cell lymphoma-2 (BCL2) family, including BCL2 and BCL2-associated X protein (BAX). Anoikis sensitization of AE was consistently observed in A549 and H292 lung cancer cells.The present study demonstrates a novel activity of AE on lung cancer cell anoikis for the first time which might lead to the development of a new strategy for lung cancer therapy.

The anoikis-sensitization activity of imperatorin, an active furanocoumarin component of Angelica dahurica root, is reported herein for the first time. The present study demonstrated that the imperatorin treatment at sub-toxic concentrations enhanced human lung cancer H23 cell apoptosis after detachment. A Western blot analysis showed that imperatorin significantly enhanced the p53 protein level, which subsequently down-regulated Mcl-1 protein and up-regulated Bax, while it had a minimal effect on Bcl-2 expression. In addition, an anchorage-independent growth assay was performed to support the anti-metastasis potential of imperatorin. Consistent with anoikis assay, imperatorin exhibited a strong inhibitory effect on the anchorage-independent growth of the cells. Further, this study demonstrated that imperatorin sensitizes anoikis in other lung cancer cells, namely, H292 and A549. Because anoikis was shown to be a critical hindrance in preventing cancer cell metastasis, the knowledge regarding such an activity and an underlying mechanism may lead to the development of this compound for a cancer therapy.

Advance therapies which effectively reduce mortality in lung cancer patients become a global health challenge nowadays. Sufficient knowledge regarding cellular and molecular basis of cancer progression and metastasis would possibly help in the development of novel and effective strategies for the treatment of this disease. This review focuses on the information regarding the role and mechanism of caveolin-1 (Cav-1) protein on anoikis in Non Small Cell Lung Cancer (NSCLC). NSCLC accounts of approximately 80% of all diagnosed lung cancer cases which patients frequently presented with metastatic disease. Like other cancers, caveolin-1 (Cav-1) has been shown to play an important role in regulating cancer-cells behaviors including apoptosis resistance and metastasis. Even though further investigations as well as in vivo experiments are required, these data are of great interest and may be beneficial to the development of lung cancer therapy.

Both caveolin-1 (Cav-1) and Mcl-1 have been implicated in the regulation of cancer cell anoikis, but their relationship and underlying mechanisms of regulation are not known. The present study demonstrated for the first time that Cav-1 regulates Mcl-1 through protein-protein interaction and inhibits its downregulation during cell anoikis in human lung cancer cells. Immunoprecipitation and immunocytochemistry studies showed that Cav-1 interacted with Mcl-1 and prevented it from degradation via the ubiquitin-proteasome pathway. Mcl-1 and Mcl-1-Cav-1 complex were highly elevated in Cav-1-overexpressing cells but were greatly reduced in Cav-1 knockdown cells. Consistent with this finding, we found that Mcl-1 ubiquitination was significantly attenuated by Cav-1 overexpression but increased by Cav-1 knockdown. Together, our results indicate a novel role of Cav-1 in anoikis regulation through Mcl-1 interaction and stabilization, which provides a new insight to the pathogenesis of metastatic lung cancer and its potential treatment.

Advances in understanding lung cancer biology and tumor markers aid clinicians in managing the disease. Cancer-associated antigen (CA)125 has garnered increasing attention in lung cancer research and may benefit the treatment and follow-up of this type of cancer. In Thai lung cancer patients, knowledge regarding ethnic differences in cancer cell biology is largely absent. We generated lung cancer cells from the pleural effusion fluids of a Thai patient and designated these as P1 cells. P1 cells were assessed for growth rate, response to chemotherapy, and the presence of tumor markers, in particular CA125 expression. Results of immunofluorescence indicated that P1 cells exhibited strong expression levels of CA125, comparable to that of established H460 lung cancer cells. Furthermore, P1 cells were analyzed for the expression of additional markers. Results revealed that H460 cells exhibited strong immunofluorescent signals from cytokeratin-19 fragments (CYFRA 21-1) and squamous cell carcinoma antigen (SCCA) while P1 presented only CYFRA 21-1 signals. We also found evidence of relative cisplatin resistance in P1 compared to the susceptibility level of established lung cancer cells. Thus, the results and methodology described in this study may aid the development of lung cancer diagnostic and therapeutic approaches and, in particular, advance understanding of ethnic differences.

Renieramycin M, has been shown to exhibit promising anticancer activity against some cancer cell lines; however, the underlying mechanism remains unknown.Renieramycin M was isolated from the blue sponge Xestospongia sp. Anticancer and antimetastatic activities of renieramycin M were investigated in human non-small cell lung cancer cells.Renieramycin M treatment caused p53 activation, which subsequently down-regulated anti-apoptotic MCL-1 and BCL-2 proteins, while the level of pro-apoptotic BAX protein was not altered. The subtoxic concentrations of renieramycin M significantly decreased invasion and migration abilities of cancer cells. In addition, this compound showed a strong inhibitory effect on anchorage-independent growth of the cells.These results reveal that renieramycin M induced lung cancer cells apoptosis through p53-dependent pathway and the compound may inhibit progression and metastasis of lung cancer cells.

In the paper titled “Long-Term Nitric Oxide Exposure Enhances Lung Cancer Cell Migration,” the Acknowledgments is corrected as follows. This work was supported by grants from the Thailand Research Fund (P. Chanvorachote) and the 90th Anniversary of Chulalongkorn University Fund (Ratchadaphiseksomphot Endowment Fund). The authors would like to thank Mr. Krich Rajprasit for his assistance in proofreading.

Background In order to gain insight into neuroprotective effects of ECa 233, a standardized extract of Centella asiatica, previously demonstrated in animal models of memory impairment induced by transient global ischemia or intracerebroventricular injection of β-amyloid, the effect of ECa 233 on neurite outgrowth of human IMR-32 neuroblastoma cell line was investigated. Methods Cells were seeded and incubated with various concentrations of ECa 233. Morphometric analysis was carried out by a measurement of the longest neurite growth of cells at 24 and 48 h. Contributing signaling pathways possibly involved were subsequently elucidated by western blot analysis. Results While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1–100 μg/ml. Western blot analysis revealed that ECa 233 significantly upregulated the level of activated ERK1/2 and Akt of the treated cells suggesting their involvement in the neuritogenic effect observed, which was subsequently verified by the finding that an addition of their respective inhibitors could reverse the effect of ECa 233 on these cells. Conclusions The present study clearly demonstrated neurite outgrowth promoting activity of ECa 233. ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed. In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.