Induced Lung Cancer Research Articles

<em>Tinospora cordifolia</em> (Willd.) Hook. f. and Thoms. and Arabinogalactan exert chemopreventive action during B(a)P induced pulmonary carcinogenesis: Studies on ultrastructural, molecular and biochemical alterations

Tinospora cordifolia (Willd.) Hook. f. and Thoms. (Tc) is a medicinal plant gaining considerable attention owing to its wide range of pharmacological properties. Though there are ample reports which have documented its biological activity in various clinical disorders, there is a paucity of information regarding its anticancer activity particularly against B(a)P induced lung cancer. So, the present study evaluates the chemopreventive potentials of aqueous extract of Tc (Aq.Tc) and its active component; Arabinogalactan (AG) against Benzo(a)pyrene [B(a)P] induced pulmonary carcinogenesis in BALB/c mice. B(a)P resulted in significant alteration in carcinogen metabolizing enzymes (CME’s), reduced glutathione and lipid peroxidation levels. B(a)P also inflicted clastogenic damage, disturbed phospholipid saturation, protein secondary structures and glycogen content. Altogether these events resulted in the alterations in cellular arrangement of pulmonary tissue, depicting the outburst of lung carcinogenesis. However, Aq.Tc and AG significantly helped to normalize the disturbed levels of CME’s and antioxidant machinery. Clastogenic changes, phospholipid saturation, protein secondary structures and glycogen content were also alleviated with Aq.Tc and AG administration. Additionally, B(a)P+ Aq.Tc and B(a)P + AG treated groups revealed the classical features of apoptosis. The results suggest that the aquatic extract of gudichi (Aq.Tc) and Arabinogalactan (AG) modulate various key processes associated with carcinogenesis and can be used as effective chemopreventive agents.

Antioxidant, antiproliferative, and apoptotic activity of thymoquinone against benzo(a)pyrene-induced experimental lung cancer.

Several studies have suggested that increased consumption of phytochemicals is a comparatively easy and practical strategy to significantly decrease the incidence of cancer. In the present study, we have reported the protective effect of a natural compound, thymoquinone (TQ) against benzo(a)pyrene (B(a)P)-induced lung carcinogenesis in Swiss albino mice. B(a)P (50 mg/kg body weight) was administered twice weekly for four successive weeks and left until 20 weeks to induce lung cancer in mice. TQ (20 mg/kg body weight) was given orally as a pretreatment and posttreatment drug to determine its chemopreventive and therapeutic effects. B(a)P-induced lung cancer-bearing animals displayed cachexia-like symptoms along with an abnormal increase in lung weight and the activities of marker enzymes adenosine deaminase, aryl hydrocarbon hydroxylase, gamma-glutamyl transpeptidase, 5'-nucleotidase and lactate dehydrogenase; tumor marker carcinoembryonic antigen levels. Furthermore, B(a)P-induced animals showed elevated levels of lipid peroxides with subsequent depletion in the antioxidant status and histological aberrations. These anomalies were accompanied by increased expressions of proliferating cell nuclear antigen and cyclin D1 in the lung sections derived from B(a)P-induced animals. On TQ treatment, all the above alterations were returned to near normalcy. Furthermore, TQ administration in B(a)P-induced animals downregulated phosphatidylinositol 3-kinase/protein kinase B signaling pathway and induced apoptosis as evidenced by a decrease in cytochrome c, proapoptotic Bax, caspase-3, and p53 with a parallel increase in antiapoptotic Bcl-2. Our present results demonstrate the potential effectiveness of TQ as an antioxidant, antiproliferative, and apoptotic agent against B(a)P-induced experimental lung tumorigenesis.

RETRACTED: Chitosan-alginate nanoparticles of cabazitaxel: Design, dual-receptor targeting and efficacy in lung cancer model

Cabazitaxel (CZT) loaded chitosan-alginate based (CSA) nanoparticles were developed with dual targeting functions of both folate receptor and epidermal growth factor receptor (EGFR) using ionic gelation technique. The chitosan-folate conjugate was synthesized, and characterized by using FTIR, NMR and Mass spectroscopy. The physicochemical parameters and morphology of all CSA nanoparticles were examined. The degree of conjugation of folic acid and cetuximab (CTXmab) was determined by UV–Visible spectroscopy and Bradford assay, respectively. Moreover, XPS analysis also supported the presence of the ligands on nanoparticles. The cellular-uptake study performed on A-549 cells demonstrated a significant enhancement in the uptake of dual-receptor targeted CSA nanoparticles than non-targeted and single-receptor targeted CSA nanoparticles. Further, CZT-loaded dual receptors targeted CSA nanoparticles also showed significantly lower IC50 values (~38 folds) than the CZT control against A-549 cells. Further, in-vivo histopathological evaluations of dual receptor-targeted CSA nanoparticles have demonstrated better safety in Wistar rats. Moreover, its treatment on the Benzo(a)pyrene (B(a)P) induced lung cancer mice model has showed the enhanced anticancer efficacy of CZT with a prolonged survival rate.

Hypoxic lung cancer cell-derived exosomal miR-21 mediates macrophage M2 polarization and promotes cancer cell proliferation through targeting IRF1

BackgroundHypoxia is the hallmark of the tumor microenvironment (TME) and plays a critical role during the progress of tumor development. A variety of microRNAs (miRNAs) transmitted by tumor-derived exosomes were involved in intercellular communication. We aimed to elucidate the precise mechanism by which tumor cell-derived exosomes promote lung cancer development by affecting macrophage polarization under hypoxic conditions.MethodsCD163 signal in tumor tissue from lung cancer patients was detected by immunohistochemical (IHC). The M2 polarization-related markers were assessed by flow cytometry and western blot. Exosomes were isolated from normoxic and hypoxic lung cancer cell culture and characterized by transmission electron microscope (TEM), dynamic light scattering (DLS), and western blot. RNA sequencing was performed to show the abnormally expressed miRNAs in exosomes from normoxic and hypoxic lung cancer cell culture. In addition, CCK-8 and clone formation assays were used to assess cell proliferation. Dual luciferase reporter assay was used to evaluate the relationship between miR-21 and IRF1. For in vivo experiment, the male nude mice were injected with H1299 cells with exosomes and miR-21 mimic treatment.ResultsFirstly, we found a strong CD163 signal in tumor tissue from lung cancer patients by IHC. Subsequently, we co-cultured lung cancer cell line H1299 with M0 macrophage THP-1 and found that H1299 in a hypoxic environment promoted THP-1 M2 polarization. PKH67 fluorescence staining experiments confirmed that exosomes of H1299 origin were able to enter THP-1 and induced M2 polarization. RNA sequencing of exosomes showed that miR-21 level was significantly higher in the hypoxic culture group compared to the normoxic group. Subsequent cellular assays showed that miR-21 inhibited the expression of IRF1 by targeting it. In addition, the overexpression of IRF1 reversed the role of miR-21 on macrophage M2 polarization. Finally, we have confirmed through animal experiments that either hypoxic environment or high miR-21 level promoted tumor progression.ConclusionsHigh miR-21 level in hypoxic environments promoted macrophage M2 polarization and induced lung cancer progression through targeting IRF1.

Frankincense essential oil nanoemulsion specifically induces lung cancer apoptosis and inhibits survival pathways

BackgroundThe volatile fraction of frankincense (Boswellia sacra) oleogum was extracted, formulated in nanoemulsion and tested against lung cancer A549 cell line. First, the gum was hydro-distilled to isolate the volatile fraction (essential oil), which was analyzed via gas chromatography to identify its major volatile constituents. Then, the oil was formulated in two water-based nanoemulsions which differ from one another in the presence of propylene glycol (PG), which is used in the formulation step as a co-surfactant. The pure essential oil as well as its major volatile compound (α-pinene), its two nanoemulsions and a reference drug (Doxorubicin) were evaluated against lung cancer A549 cell lines and WI-38 normal lung cells. The evaluation included cytotoxicity (MTT and IC50), apoptosis (flow cytometric analysis) in addition to genetic assessments for some intrinsic and extrinsic genes relevant to apoptosis and survival pathways.ResultsChromatographic analysis of frankincense essential oil revealed that α-pinene is the major volatile compound which constituent about 60% of that oil. Emulsification of the oil using the low energy technique gave nanoemulsions having major intense particles population (85–90%) with z-average diameter below 20.0 nm. Frankincense oil nanoemulsion fabricated with (PG) showed the best cytotoxic activity toward lung cancer A549 cell compared to PG-free nanoemulsion, α-pinene and the reference drug doxorubicin, along different incubation periods. Flow cytometric analysis also indicated that PG-containing nanoemulsion can induce cancer cells toward apoptosis better than the other formula and the pure oils. The same nanoemulsion was found to upregulate the pro-apoptotic genes [DR5, FAAD, Caspase 8 (Cas8), p53, and Bax] and downregulate the anti-apoptotic and reoccurrence genes (Bcl-2, NF-kB, and STAT-3). Most importantly, the PG-containing nanoemulsion had the least cytotoxic effect on the normal WI-38 lung cells.ConclusionsThese results point out to the potentials of frankincense essential oil (rich in α-pinene) and its PG-nanoemulsion as a promising adjuvant from plant-source to potentiate the activity of the systematic anti-lung cancer drugs.

ITCH facilitates proteasomal degradation of TXNIP in hypoxia- induced lung cancer cells.

BackgroundLung cancer (LC) is one of the most common cancers and a leading cause of cancer‐related deaths worldwide. In many pathological conditions, particularly in the tumor microenvironment, cells and tissues frequently exist in a hypoxic state.Here, we evaluated Itchy E3 ubiquitin protein ligase (ITCH) expression in LC cells following hypoxia treatment.MethodsLC cell lines were treated with hypoxic condition. Cell migration, invasion, inflammation, reactive oxygen species (ROS) production, and apoptosis of LC cells were determined by wound healing assay, Transwell invasive assay, ELISA, DCFH‐DA staining, and flow cytometry, respectively. qPCR and WB were used to determine the expression of ITCH and TXNIP. Co‐IP was performed to assess the interaction between ITCH and TXNIP.ResultsITCH expression was downregulated in LC cells under hypoxic conditions. Next, LC cells were subjected to hypoxic conditions and changes in cell viability and metastasis were determined. Hypoxic conditions resulted in increased migration and invasion abilities of LC cells. Intracellular reactive oxygen species (ROS) production, inflammation, and apoptosis were also promoted by hypoxia. We found that ITCH overexpression led to the proteasomal degradation of thioredoxin‐interacting protein (TXNIP), whereas the expression of the ITCH C830A mutant did not affect TXNIP levels in LC cells. The gain‐of‐function experiment demonstrated that migration, invasion, ROS generation, inflammation, and apoptosis of hypoxia‐conditioned LC cells were ameliorated by ITCH overexpression, whereas the ITCH C830A mutant did not cause any changes in these phenotypes. Furthermore, the contribution of TXNIP knockdown and ITCH overexpression to the hypoxia‐induced features in LC cells with ITCH C830A was found to be similar.ConclusionOur results suggest a novel mechanism underlying the changes in ITCH‐mediated malignant phenotypes of hypoxia‐conditioned LC cells via TXNIP.

Connexin 43 overexpression induces lung cancer angiogenesis in vitro following phosphorylation at Ser279 in its C-terminus.

Blocking angiogenesis can inhibit tumor growth and metastasis. However, the mechanism underlying regulation of lung cancer angiogenesis remains unclear. The gap junction protein connexin 43 (Cx43) is implicated in angiogenesis. The aim of the present study was to determine the role of Cx43 in angiogenesis in vitro and its signaling pathways. Human pulmonary microvascular endothelial cells were transfected with Cx43-targeting siRNA or Cx43-overexpressing recombinant plasmid vector. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to determine Cx43, zonula occludens-1 (ZO-1), E-cadherin, β-catenin, von Willebrand factor (vWF), and plasminogen activator inhibitor-1 (PAI-1) mRNA and protein expression levels, respectively. Tyr265, Ser279, Ser368, and Ser373 phosphorylation levels in the C-terminus of Cx43 and intracellular and membranal Cx43 contents were determined using western blotting. Additionally, immunofluorescence, tube formation, Cell Counting Kit-8, and Transwell migration assays were performed. The results revealed that compared with that in the control samples, Cx43, ZO-1, E-cadherin, β-catenin, vWF, and PAI-1 mRNA and protein expression were significantly increased in the Cx43 overexpression group and significantly decreased in the Cx43-knockdown group. Moreover, the phosphorylation level of Ser279 as well as cell proliferation and migration rates were markedly increased in the Cx43 overexpression group, and tube formation revealed that the potential of angiogenesis was also increased. Conversely, in the Cx43-knockdown group, the phosphorylation level of Ser279 and cell proliferation and migration rates were reduced, and the potential of angiogenesis was greatly impaired. Under Cx43 overexpression, membranal Cx43 content was significantly increased, whereas under Cx43 knockdown, it was significantly reduced. Therefore, Cx43 overexpression could induce pulmonary angiogenesis in vitro by promoting cell proliferation and migration and activating ZO-1, E-cadherin, β-catenin, vWF, and PAI-1. This may be achieved by promoting phosphorylation and activation of the intracellular signal site Ser279 at the C-terminus of Cx43.

Long-chain non-coding RNA LINC00473 antagonist with liposomal nanoparticles as carrier targets for Keap1/Nrf2/ARE signaling pathway to promote lung cancer cell apoptosis

There is a relationship between abnormal expression of LncRNA LINC00473 and lung cancer cell apoptosis. This study mainly explored the role and mechanism of LncRNA LINC00473 in lung cancer cell apoptosis. Lung cancer cell line A549 was taken and study groups were set as follows; negative control (NC) group, si-con group, si-LINC00473 group, miR-634 group, si-LINC00473+anti-miR-con group, and si-LINC00473+antimiR-634 group. The expressions of LncRNA LINC00473 and miR-634 were observed and changes of cell proliferation and apoptosis-related indicators (Cyclin D1, Cleaved-caspase-3, cell survival rate, apoptosis rate) after interfering with expression of LncRNA LINC00473 and miR-634 respectively. Compared to HT-ori3 cells, the expression of LncRNA LINC00473 was higher while that of miR-634 was lower. Compared with the other two groups, the expressions of LncRNA LINC00473, Cyclin D1 and cell survival rate were lowest in the si-LINC00473 group, while the expression and apoptosis rate of Cleaved-caspase-3 were highest. Compared to the other two groups, the expression of miR-634 and Cleaved-caspase-3 and apoptosis rate in the miR-634 group were highest, and the cell survival rate of Cyclin D1 was lowest (p < 0.05). LncRNA LINC00473 gene sequence and miR-634 gene 3′UTR 641–678 sequence have specific binding regions, and miR-634 is the target gene for LncRNA LINC00473. The fluorescence intensity of mutant plasmid was markedly higher than that of wild-type plasmid (p<0.05). Compared with si-con group, the si-LINC00473 and si-LINC00473+anti-miR-con groups had higher miR-634, Cleaved-caspase-3 and apoptosis rates, while the Cyclin D1, cell survival rate, Keap1, p-Nrf2, and P-ARE expressions were low, but the si-LINC00473+antimiR-634 group was opposite. The LncRNA LINC00473 targets the RNA binding to the miR-634, thereby up-regulating the miR-634 expression and down-regulating the Keap1 expression.The LncRNA LINC00473 also inhibits the Nrf2, ARE phosphorylation and Cyclin D1 expression, and also promotes Cleaved-caspase-3 expression, ultimately inducing lung cancer cell apoptosis and inhibiting cell proliferation.

Biosynthesized CdS Nanoparticle Induces ROS-dependent Apoptosis in Human Lung Cancer Cells.

The World Health Organization (WHO) estimated that the number of cancer-related deaths was 9.6 million in 2018 and 2.09 million deaths occurred by lung cancer. The American Institute for Cancer Research (AICR) also observed gender preferences in lung cancer, common in men than women. Since the past decade, nanoparticles have now been widely documented for their anti-cancer properties, which signifies that the development of nanotechnology would be a future diagnosis and treatment strategy for lung cancer. The current study aimed to investigate the role of biosynthesized CdS nanoparticles (CdS NPs) in lung cancer cells (A549). Therefore, whether the CdS NP induces lung cancer cell death and the underlying mechanism is yet to be elucidated. Literature was searched from various archives of biomedical and life science journals. Then, CdS NPs were biosynthesized and characterized by traditional and cutting-edge protocols. The CdS NP-mediated cell death was elucidated following standard protocols. CdS NPs induced cytotoxicity towards A549 cells in a dose-dependent manner. However, such a death mechanism does not go through necrosis. Intracellular reactive oxygen species (ROS) accumulation and mitochondrial membrane depolarization demonstrated that cell death is associated with intracellular ROS production. Furthermore, increased sub-G1 population, Bax expression, and decreased Bcl-2 expression revealed that the death was caused by apoptosis. CdS NPs promote apoptosis-mediated lung cancer cell death through ROS production.

Does pulmonary fibrosis increase lung cancer risk?

Abstract Background The concepts in lung fibrosis and increased lung cancer risk have arisen from clinical observations of the concurrent findings during the autopsy of lung cancer and pulmonary fibrosis. Scar carcinoma attributes the origin of carcinomas to abutting areas of scarring. The association between fibrosis and lung cancer risk immediately raises a query: whether lung fibrosis induces lung cancer? Aim The study was done to elucidate the incidence, risk factors, and clinical characteristics of lung cancer in diffuse lung fibrosis. Patients and methods A cohort study was done based on a prospective group of 50 patients with diffuse lung fibrosis to evaluate the incidence of lung cancer, as well as a retrospective group of 50 patients with lung cancer to search for background of pulmonary fibrosis between 2019 and 2020. Demographic, radiographic, and histologic characteristics were reviewed for both groups. Results Among 50 patients with interstitial lung disease, lung cancer developed in 2/50 (4%) during the follow-up, whereas in the retrospective group, 8/50 (16%) were diagnosed as having lung cancer with diffused lung fibrosis between 2019 and 2020. Lung cancer risk factors were male sex, smoking, and occupational dust exposure. Squamous carcinoma followed by adenocarcinoma was the commonest histologic form. Conclusion Evidence shows an association between interstitial lung diseases and lung cancer emergence. Epidemiological evidence differs owing to the various study designs and populations.

Anticancer activity of D-LAK-120A, an antimicrobial peptide, in non-small cell lung cancer (NSCLC)

Non-small cell lung cancer (NSCLC) is a major cause of global cancer mortalities and accounts for approximately 80–85% of reported lung cancer cases. Conventional chemotherapeutics show limited application because of poor tumor selectivity and acquired drug resistance. Antimicrobial peptides (AMPs) have gained much attention as potential anticancer therapeutics owing to their high potency and high target selectivity and specificity with limited scope for drug resistance. In this study, D-LAK (D-LAK-120A), a cationic AMP, was evaluated for its anticancer efficacy in various NSCLC cell lines. D-LAK peptide demonstrated enhanced cytotoxicity in A549, H358, H1975, and HCC827 cell lines with inhibitory concentrations between 4.0 and 5.5 μM. An increase in the lactate dehydrogenase (LDH) levels and propidium iodide (PI) uptake across compromised membrane suggested membranolytic activity as an inhibition pathway. In addition, we found D-LAK induced lung cancer cell apoptosis and arrested cells in the S phase (DNA synthesis) of cell cycle. Moreover, a decreased mitochondrial membrane potential and elevated ROS levels were observed after D-LAK treatment, suggesting induction of mitochondria-mediated apoptosis. Additionally, D-LAK inhibited single cell proliferation and cancer cell migration in vitro. The tumor reduction observed in the 3D spheroid assay further suggests the potential use of D-LAK as an anticancer agent for NSCLC treatment. Our results postulate innovative insights on the anticancer mechanism of D-LAK, which may contribute to its further development into preclinical studies and a potential therapeutic.

Abstract 3839: RASSF1C and tumor microenvironment

Abstract Introduction: Tumor microenvironment (TME) plays a vital role in tumor invasion and metastasis and provides a rich environment for identifying novel therapeutic targets. The TME landscape consists of extracellular matrix (ECM) and stromal cells. ECM is a major component of TME that mediates interaction between cancer cells and stromal cells to promote invasion and metastasis. We have shown in published work that RASSF1C promotes cancer stem cell development, migration, and drug resistance, in part, by promoting EMT through a mechanism that involves up-regulation of the PIWIL1-piRNA gene axis. Interestingly, one of the target genes identified by our microarray study to be upregulated by RASSF1C is P4HA2. In cancer, P4H2A is vital for collagen posttranslational modification and folding. This leads to formation of a stiff ECM and induction of cancer stem cell marker gene expression, resulting in metastatic dissemination. High expression of P4HA2 is associated with significantly lower survival of lung cancer patients. Thus, RASSF1C could promote tumor cell ECM remodeling to induce lung cancer cell stemness, invasion and metastasis by up-regulating a novel PIWIL1-P4HA2 gene axis expression. Methods: Analysis of a microarray study using H1299 cells over-expressing RASSF1C or control vector backbone identified P4HA2 as a RASSF1C gene target. RT-PCR, immunoblots, and immunofluorescence techniques were used to confirm P4HA2 gene expression using specific primers and antibodies in cells over-expressing RASSF1C or PIWIL1 along with control cells. Kaplan-Meier analysis was performed to determine the relationship of P4HA2 expression to lung adenocarcinoma patient survival using data from The Cancer Genome Atlas (TCGA). Results: Analysis of microarray data from H1299 cells over-expressing RASSF1C or control vector backbone shows that RASSF1C up-regulates P4HA2 gene expression 2-fold. The up-regulation of P4HA2 expression by RASSF1C was confirmed by RT-PCR, immunoblots, and immunofluorescence using P4HA2 gene specific primers and antibodies. Further, H1299 cells over-expressing PIWIL1 show increased P4HA2 gene expression on immunoblots. Kaplan-Meier analysis shows that high P4HA2 expression in lung adenocarcinoma patients negatively correlates with patient survival. Thus, our findings suggest the hypothesis that RASSF1C may promote lung cancer cell ECM remodeling to induce lung cancer cell stemness, invasion and metastasis, in part, by up-regulating a novel pathway involving a PIWIL1-P4HA2 gene axis. Conclusions: Investigating the role of a RASSF1C-PIWIL1-P4HA2 gene axis in ECM remodeling in lung cancer cells will provide important mechanistic information that could, in turn, lead to identification of useful biomarkers for lung cancer prognosis and targets for therapy. Citation Format: Yousef G. Amaar, Mark E. Reeves. RASSF1C and tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3839.

Abstract 228: Depletion of stem-loop binding protein and elevation of polyadenylation of histone 3.1 contribute to malignant cell transformation induced by bisphenol A

Abstract Introduction: Bisphenol A (BPA) is a pervasive chemical exposure in the US and has been linked to adverse health effects in humans and mainly identified as an endocrine disruptor. However, recent studies suggests that all plastics including our daily usages for food/drink containers may leach chemicals if they are scratched or heated or stored long term and strongly suggests that at certain exposure levels of BPA in these products, may cause cancers including breast, lung, and ovarian cancers in humans. Although there are few studies about breast, lung and ovarian cancers, with BPA exposure there is an urgent need to focus on the BPA’s carcinogenic activity in the lung. Because lung cancers can progress to life threatening events and short life span once it is diagnosed, identifying the mechanism that specifically inhibits/prevents lung cancers invasion and related morbidity is of tremendous importance for reducing mortality. Therefore, in this study, we investigated the carcinogenicity of BPA using human bronchial epithelial cells. Experimental methods: To determine whether BPA alters canonical histone mRNA processing in vitro, we measured levels of polyadenylation (polyA) of canonical histone (H)3.1 mRNA as well as protein expression of Stem-loop binding protein (SLBP), and H3 protein in human bronchial epithelial BEAS-2B cells exposed to various concentrations of BPA for 96 h. Furthermore, BEAS-2B cells were chronically exposed to low doses of BPA, the cell transformation assay was performed in an anchorage-independent growth and colony formation was assessed. Arsenic was used as a positive control because our laboratory previously demonstrated that arsenic-induced polyA of H3.1 mRNA, by depletion of SLBP in vitro and that polyA of H3.1 mRNA enhanced tumor formation in nude mice. Results: Our results showed that exposure of BEAS-2B cells to BPA depleted SLBP, induced polyA of canonical histone mRNAs, and upregulated H3 protein. The results from real-time quantitative PCR showed that BPA exposure increased level of polyA H3.1 mRNA. Furthermore, our findings from an anchorage-independent growth assay also indicate that exposure of BEAS-2B cells to low doses of BPA induced malignant cell transformation. Conclusions: The present study demonstrated that chronic exposure of the cells to BPA causes malignant cell transformation, indicating that BPA is a potent human carcinogen to induce lung cancer. Our study also demonstrated that the loss of SLBP and gain of polyA H3.1 mRNA play an important role in BPA-induced carcinogenesis. Citation Format: Arul Veerappan, Zhou Zhang, Max Costa. Depletion of stem-loop binding protein and elevation of polyadenylation of histone 3.1 contribute to malignant cell transformation induced by bisphenol A [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 228.

Air pollution particles hijack peroxidasin to disrupt immunosurveillance and promote lung cancer.

Although fine particulate matter (FPM) in air pollutants and tobacco smoke is recognized as a strong carcinogen and global threat to public health, its biological mechanism for inducing lung cancer remains unclear. Here, by investigating FPM's bioactivities in lung carcinoma mice models, we discover that these particles promote lung tumor progression by inducing aberrant thickening of tissue matrix and hampering migration of antitumor immunocytes. Upon inhalation into lung tissue, these FPM particles abundantly adsorb peroxidasin (PXDN) - an enzyme mediating type IV collagen (Col IV) crosslinking - onto their surface. The adsorbed PXDN exerts abnormally high activity to crosslink Col IV via increasing the formation of sulfilimine bonds at the NC1 domain, leading to an overly dense matrix in the lung tissue. This disordered structure decreases the mobility of cytotoxic CD8+ T lymphocytes into the lung and consequently impairs the local immune surveillance, enabling the flourishing of nascent tumor cells. Meanwhile, inhibiting the activity of PXDN abolishes the tumor-promoting effect of FPM, indicating the key impact of aberrant PXDN activity on the tumorigenic process. In summary, our finding elucidates a new mechanism for FPM-induced lung tumorigenesis and identifies PXDN as a potential target for treatment or prevention of the FPM-relevant biological risks.

In-vivo dose determination in a human after radon exposure: proof of principle

Radon-222 is pervasive in our environment and the second leading cause of lung cancer induction after smoking while it is simultaneously used to mediate anti-inflammatory effects. During exposure, radon gas distributes inhomogeneously in the body, making a spatially resolved dose quantification necessary to link physical exposure conditions with accompanying risks and beneficial effects. Current dose predictions rely on biokinetic models based on scarce input data from animal experiments and indirect exhalation measurements of a limited number of humans, which shows the need for further experimental verification. We present direct measurements of radon decay in the abdomen and thorax after inhalation as proof of principle in one patient. At both sites, most of the incorporated radon is removed within ~ 3 h, whereas a smaller fraction is retained longer and accounts for most of the deposited energy. The obtained absorbed dose values were 1.5 pm 0.3 µGy (abdomen, radon gas) and 1.1 pm 0.4 µGy (thorax, radon and progeny) for a one-hour reference exposure at a radon activity concentration of 55 kBq m−3. The accumulation of long-retained radon in the abdomen leads to higher dose values at that site than in the thorax. Contrasting prior work, our measurements are performed directly at specific body sites, i.e. thorax and abdomen, which allows for direct spatial distinction of radon kinetics in the body. They show more incorporated and retained radon than current approaches predict, suggesting higher doses. Although obtained only from one person, our data may thus represent a challenge for the barely experimentally benchmarked biokinetic dose assessment model.

Experimental and Theoretical Insights on Chemopreventive Effect of the Liposomal Thymoquinone Against Benzo[a]pyrene-Induced Lung Cancer in Swiss Albino Mice.

PurposeThymoquinone (TQ), a phytoconstituent of Nigella sativa seeds, has been studied extensively in various cancer models. However, TQ’s limited water solubility restricts its therapeutic applicability. Our work aims to prepare the novel formulation of TQ and assess its chemopreventive potential in chemically induced lung cancer animal model.MethodsThe polyethylene glycol coated DOPE/CHEMS incorporating TQ-loaded pH-sensitive liposomes (TQPSL) were prepared and characterized. Mice were exposed to benzo[a]pyrene (BaP) thrice a week for 4 weeks to induce lung cancer. TQPSL was administered three times a week for 21 weeks, starting 2 weeks before the first dose of BaP.ResultsThe prepared TQPSL revealed 85% entrapment efficiency with 128 nm size and −19.5 mv ζ-potential showing high stability of the formulation. The pretreatment of TQPSL showed the recovery in BaP-modulated relative organ weight of lungs, cancer marker enzymes, and antioxidant enzymes in the serum. The histopathological analysis of the tissues showed that TQPSL protected the malignancy in the lungs. The flow cytometry data revealed the induction of apoptosis and decreased intracellular ROS by TQPSL. Molecular docking was performed to predict the TQ’s affinity for eight possible anticancer drug targets linked to lung cancer etiology. The data assisted to identify the serine/threonine-protein kinase BRAF as the most suitable target of TQ with binding energy −6.8 kcal/mol.ConclusionThe current findings demonstrated the potential of TQPSL and its possible therapeutic targets of lung cancer. To our knowledge, this is the first research to outline the development of TQ formulation against lung cancer considering its low solubility as well as pulmonary delivery challenges.

USP15 negatively regulates lung cancer progression through the TRAF6-BECN1 signaling axis for autophagy induction

TNF receptor-associated factor 6 (TRAF6)-BECN1 signaling axis plays a pivotal role in autophagy induction through ubiquitination of BECN1, thereby inducing lung cancer migration and invasion in response to toll-like receptor 4 (TLR4) stimulation. Herein, we provide novel molecular and cellular mechanisms involved in the negative effect of ubiquitin-specific peptidase 15 (USP15) on lung cancer progression. Clinical data of the TCGA and primary non-small cell lung cancer (NSCLC) patients (n = 41) revealed that the expression of USP15 was significantly downregulated in lung cancer patients. Importantly, USP15-knockout (USP15KO) A549 and USP15KO H1299 lung cancer cells generated with CRISPR-Cas9 gene-editing technology showed increases in cancer migration and invasion with enhanced autophagy induction in response to TLR4 stimulation. In addition, biochemical studies revealed that USP15 interacted with BECN1, but not with TRAF6, and induced deubiquitination of BECN1, thereby attenuating autophagy induction. Notably, in primary NSCLC patients (n = 4) with low expression of USP15, 10 genes (CCNE1, MMP9, SFN, UBE2C, CCR2, FAM83A, ETV4, MYO7A, MMP11, and GSDMB) known to promote lung cancer progression were significantly upregulated, whereas 10 tumor suppressor genes (FMO2, ZBTB16, FCN3, TCF21, SFTPA1B, HPGD, SOSTDC1, TMEM100, GDF10, and WIF1) were downregulated, providing clinical relevance of the functional role of USP15 in lung cancer progression. Taken together, our data demonstrate that USP15 can negatively regulate the TRAF6-BECN1 signaling axis for autophagy induction. Thus, USP15 is implicated in lung cancer progression.

Atractylenolide II inhibits tumor-associated macrophages (TAMs)-induced lung cancer cell metastasis

Objective M2-like tumor-associated macrophages (TAMs) play a crucial role in promoting tumor proliferation, angiogenesis, and metastasis. In the current study, we investigated the relationship between macrophage polarization and the antitumor effect of Atractylenolide II (AT-II) in lung cancer cells. Materials and methods Cell viability, migration, and invasion were determined by MTT assay, wound healing assay, and transwell assay, respectively. Flow cytometry analysis showed the percentage of CD206+ cells. Gene expression was determined by real-time PCR, western blotting, and immunofluorescence staining. Lewis lung carcinoma mouse xenograft and metastasis models were used to examine the effects of AT-II on lung cancer in vivo. Results AT-II (2.5 and 5 µM) did not cause significant inhibition of A549 cell viability but markedly inhibited IL-4/IL-13-induced M2-like polarization, evidenced by the decreased expression of the M2 surface marker CD206, down-regulation of specific M2-marker genes (Arg-1, IL-10 and TGF-β) as well as inhibition of M2 macrophages-mediated invasion and migration of A549 cells. In addition, AT-II inhibited IL-4/IL-13-induced activation of the STAT6 signaling pathway that is vital in the M2-like polarization of macrophages. In animal models, administration of AT-II (50 mg kg−1, i.g., QD for 21 days) significantly inhibited tumor growth, reduced pulmonary metastatic nodules, and down-regulated the percentages of M2 macrophages (F4/80+ and CD206+) in total macrophages (F4/80+) in tumor tissues and pulmonary metastatic nodules. Conclusions AT-II effectively inhibits M2-like polarization, thereby inhibiting lung cancer cell metastasis both in vivo and in vitro, revealing a novel potential strategy for the antitumor effect of AT-II.

Influence of Artocarpus heterophyllus in benzopyrene induced lung cancer

Lung cancer a deadliest cancer worldwide and the most common cancer in the whole of Asia. The present study focused on the protective efficacy of Artocarpus heterophyllus in benzopyrene induced lung toxicity in C57BL/6 Mice. Mice were orally induced with benzopyrene and treated with hydroethanolic extract of spermoderm of Artocarpus heterophyllus (HESA) at a dose of (50mg/kg.b.wt and 100mg/kg.b.wt). The oral administration of HESA effectively suppressed lung cancer as revealed by the decrease in the extent of haematological parameters with concomitant decrease in the activities of serum markers. Significantly protein expressions of p53 and Bcl2 in the lung tissue were further confirmed that coadministration of the extract treatment however downregulated their expressions by decreasing the marker of positively stained cells and restored the histopathological alteration of lung tissue. Our results suggest that HESA can be used as a protective agent against benzopyrene induced lung cancer.

Photodynamic Therapy with an AlPcS4Cl Gold Nanoparticle Conjugate Decreases Lung Cancer’s Metastatic Potential

Cancer metastasis and the risk of secondary tumours are the leading causes of cancer related death, and despite advances in cancer treatment, lung cancer remains one of the leading causes of death worldwide. A crucial characteristic of metastases is cell invasion potential, which is mainly determined by cell motility. Photodynamic therapy (PDT), known for its minimally invasive cancer treatment approach, has been extensively researched in vitro and is currently being developed clinically. Due to their physicochemical and optical properties, gold nanoparticles have been shown to increase the effectivity of PDT by increasing the loading potential of the photosensitizer (PS) inside cancer cells, to be biocompatible and nontoxic, to provide enhanced permeability and retention, and to induce lung cancer cell death. However, effects of gold nano phototherapy on lung cancer metastasis are yet to be investigated. The aim of this in vitro study was to determine the inhibitory effects of PS-gold nano bioconjugates on lung cancer metastasis by analysing cell proliferation, migration, cell cycle analysis, and extracellular matrix cell invasion. The findings indicate that nano-mediated PDT treatment of lung cancer prevents lung cancer migration and invasion, induces cell cycle arrest, and reduces lung cancer proliferation abilities, elaborating on the efficacy of the nano-mediated PDT treatment of lung cancer.