Pyrene-induced Lung Cancer Research Articles

Formulation and evaluation of cetuximab functionalized phospholipid modified nanocrystals of paclitaxel for non-small cell lung cancer therapy

Present work aims to prepare Soluplus stabilized, phospholipid-modified, and cetuximab-conjugated paclitaxel nanocrystals (NCs) as stable nanocarriers for targeted drug delivery. The NCs, prepared using concurrent antisolvent precipitation cum cold crystallization method followed by probe sonication, were found to be monodispersed particles with sub-200 nm size. The microscopic analysis uncovered rod and spherical anisotropy for Soluplus stabilized (PTX-NCs) and phospholipid modified (Lipid/PTX-NCs) nanocrystals, respectively. The formation of amorphous PTX-NCs and subsequent coating with phospholipid was confirmed by solid-state characterization using differential scanning calorimetry (DSC), X-ray diffraction (XRD), and Fourier transform Infrared Spectroscopy (FTIR). X-ray Photoelectron Spectroscopic (XPS) analysis, indicated successful conjugation of cetuximab on NCs surface. Lipid coating rendered a sustained drug release behaviour to NCs at physiological pH. In vitro cell line studies confirmed the improved cellular internalization and better apoptosis induction capability of NCs, consequently resulting in enhanced efficacy of PTX against A549 cancer cells. Moreover, in Benzo[a] pyrene-induced lung cancer model, Cmab/Lipid/PTX-NCs showed significant improvement in tumor inhibition potential in comparison to pure PTX. The prepared Cmab/Lipid/PTX-NCs also exhibited improved pharmacokinetics performance, avoided off-target distribution, and showed a reduction in systemic toxicity. The findings of this study indicate the promising potential of the prepared cetuximab-functionalized phospholipid-coated paclitaxel nanocrystals in lung cancer therapy.

Circ0087385 promotes DNA damage in benzo(a)pyrene-induced lung cancer development by upregulating CYP1A1.

Increasing environmental genotoxic chemicals have been shown to induce epigenetic alterations. However, the interaction between genetics and epigenetics in chemical carcinogenesis is still not fully understood. Here, we constructed an in vitro human lung carcinogenesis model (16HBE-T) by treating human bronchial epithelial cells with a typical significant carcinogen benzo(a)pyrene (BaP). We identified a novel circular RNA, circ0087385, which was overexpressed in 16HBE-T and human lung cancer cell lines, as well as in lung cancer tissues and serum exosomes from lung cancer patients. The upregulated circ0087385 after exposure to BaP promoted DNA damage in the early stage of chemical carcinogenesis and affected the cell cycle, proliferation, and apoptosis of the malignantly transformed cells. Overexpression of circ0087385 enhanced the expression of cytochrome P450 1A1 (CYP1A1), which is crucial for metabolically activating BaP. Interfering with circ0087385 or CYP1A1 reduced the levels of ultimate carcinogen benzo(a)pyrene diol epoxide (BPDE) and BPDE-DNA adducts. Interfering with CYP1A1 partially reversed the DNA damage induced by high expression of circ0087385, as well as decreased the level of BPDE and BPDE-DNA adducts. These findings provide novel insights into the interaction between epigenetics and genetics in chemical carcinogenesis which are crucial for understanding the epigenetic and genetic toxicity of chemicals.

In Vivo Cancer Microenvironment Responsive Glycan Receptor-Targeted Nanoparticles for Gemcitabine Delivery to Benzo[a]pyrene-Induced Lung Cancer Model.

Targeted gemcitabine (GEB) loaded 5-N-acetyl-neuraminic acid (Neu5Ac) assembled chitosan nanoparticles (CA-NPs) were formulated by ionotropic gelation process and evaluated for physicochemical and morphological characterization, in vitro and in vivo studies in A-549 cells and lung cancer mice model, respectively. The mean diameter of GEB-CA-Neu5Ac-NPs determined by dynamic light scattering was 161.16 ± 7.70 nm with a polydispersity index (PDI) value of 0.303 ± 0.011 and its zeta potential and entrapment efficiency (%EE) were 40.3 ± 3.45 mv and 66.11 ± 1.94%, respectively. The in vitro cellular uptake studies showed that glycan receptor-targeted nanoparticles deliver significantly more amount (p < 0.001) of GEB into the A-549 lung cancerous cells than non-targeted nanoparticles. The cytotoxicity study using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay clearly demonstrated that GEB-CA-Neu5Ac-NPs have lower IC50 value (6.39 ± 3.78 µg/ml) than others groups that showed that the greater lung cancerous cells inhibition potential of targeted nanoparticles. The in vivo biodistribution of the GEB-loaded 5-N-acetyl-neuraminic acid conjugated chitosan nanoparticles was revealed that targeted nanoparticles showed higher accumulation and retention for an extended period of time due to the active targeting ability of Neu5Ac to glycan receptors. Histopathological examination showed significant recovery in the physiological architecture upon administration of targeted nanoparticles. The glycan receptor-targeted nanoparticles treated groups showed a significant decline in the number of metastatic lung epithelial cells, as compared to the untreated positive control group (p < 0.001) confirming higher anticancer efficacy of the GEB-CA-Neu5Ac-NPs.

Benzo(A)Pyrene-Induced Lung Cancer: Chemo Protective Effect of Coronarin D in Swiss Albino Mice.

Lung cancer is considered one of the most prevalent cancers worldwide and also has a high death rate. The prevalence of lung cancer is high in developed countries than in developing countries due to the lifestyle changes and quality of air. Coronarin D is a diterpene, which is isolated from the Hedychium coronarium. It demonstrated several pharmacological properties such as anti-allergic, anti-inflammatory, antimicrobial, and anticancer activities. In the current investigation, the potential of Coronarin D on the B(a)P-induced lung cancer was studied in the experimental mice model. The B(a)P-administrated animals exhibited a reduced level of immune cells, IgG, IgM, immune complexes, SOD, and CAT. The B(a)P-administrated animals expressed high levels of IgA, LPO, xenobiotic markers, tissue marker, tumor marker, and proinflammatory cytokines. On treatment with Coronarin D, the level of neutrophils, lymphocytes, leucocytes, and absolute neutrophils was elevated in the B(a)P-administered mice. The immune complex was augmented in the Coronarin D-treated animals in comparison with B(a)P-treated mice. The level of IgG and IgM was increased, whereas the level of IgA was reduced in the Coronarin D-treated animals. The level of LPO was downregulated, whereas the level of SOD and CAT was upregulated in Coronarin D-treated animals. The expression level of xenobiotic markers, tissue marker, tumor marker, and proinflammatory cytokines was reduced in the Coronarin D-treated animals. The histopathological results revealed that lung tissues of Coronarin D-treated animals had less alveolar damage with decreased hyperplasia. These findings suggest that the Coronarin D can be utilized as a potent chemopreventive agent for treating lung cancer in the future.

Vanillic acid attenuates cell proliferation, xenobiotic enzyme activity, and the status of pulmonary mitochondrial enzymes in lung carcinoma.

The purpose of the study is to determine the anti-proliferative and mitochondrial status of benzo(a)pyrene-induced lung cancer in Swiss albino mice, as well as the modulatory effect of vanillic acid on it. B(a)P had altered levels of lysosomal enzymes, xenobiotic-metabolizing enzymes, cell proliferation, inflammation, and mitochondrial abnormalities, whereas treatment with VA treatment significantly reversed the aforementioned activities. According to the findings, VA greatly reduces lung carcinogenesis by restoring antioxidants and xenobiotic-enzyme levels, consequently proving to be an anti-proliferative and anti-inflammatory drug against lung cancer in mice. PRACTICAL APPLICATIONS: As we all know, lung cancer is on the rise all over the world. A recent study demonstrated that vanillic acid protects against B(a)P in experimental mice. According to the findings, VA considerably suppresses lung carcinogenesis by restoring lysosomal enzyme levels, xenobiotic-metabolizing enzyme levels, and mitochondrial activities, effectively functioning as an anti-proliferative and anti-inflammatory therapy against lung cancer. According to the most recent study, vanillic acid can be used as a defensive medicine in the treatment of lung cancer.

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.

CXCL13 in Cancer and Other Diseases: Biological Functions, Clinical Significance, and Therapeutic Opportunities

The development of cancer is a multistep and complex process involving interactions between tumor cells and the tumor microenvironment (TME). C-X-C chemokine ligand 13 (CXCL13) and its receptor, CXCR5, make crucial contributions to this process by triggering intracellular signaling cascades in malignant cells and modulating the sophisticated TME in an autocrine or paracrine fashion. The CXCL13/CXCR5 axis has a dominant role in B cell recruitment and tertiary lymphoid structure formation, which activate immune responses against some tumors. In most cancer types, the CXCL13/CXCR5 axis mediates pro-neoplastic immune reactions by recruiting suppressive immune cells into tumor tissues. Tobacco smoke and haze (smohaze) and the carcinogen benzo(a)pyrene induce the secretion of CXCL13 by lung epithelial cells, which contributes to environmental lung carcinogenesis. Interestingly, the knockout of CXCL13 inhibits benzo(a)pyrene-induced lung cancer and azoxymethane/dextran sodium sulfate-induced colorectal cancer in mice. Thus, a better understanding of the context-dependent functions of the CXCL13/CXCR5 axis in tumor tissue and the TME is required to design an efficient immune-based therapy. In this review, we summarize the molecular events and TME alterations caused by CXCL13/CXCR5 and briefly discuss the potentials of agents targeting this axis in different malignant tumors.

Bromelain and Olea europaea (L.) leaf extract mediated alleviation of benzo(a)pyrene induced lung cancer through Nrf2 and NFκB pathway.

Lung cancer is the most aggressive as well as deadly form of cancer and most of the lung cancer cases are involved in direct smoking or passive smoking. Oxidative stress and pulmonary inflammation regulated by some transcription factors like Nrf2, NF-κB etc. play important roles in lung cancer. Various combinations of therapies are currently attributed to lung cancer treatment. A plethora of evidence supports that the consumption of plant-derived foods can prevent chronic diseases like cancer. Leaves of olive (Olea europaea L.) are rich in phenolic compounds which are having antioxidant and anti-inflammatory property. Also, bromelain from pineapple juice and from pineapple stem is a potent anti-inflammatory agent. We took a pragmatic approach to prevent carcinogenesis by supplementing the combination of these two extracts. In this study, we have tried to evaluate the amelioration of various hallmarks associated with benzo(a)pyrene-induced lung carcinogenesis upon the combinatorial treatment of ethanolic olive leaf extract (EOLE) and bromelain. We have studied the role of EOLE in amelioration of BaP-induced oxidative stress in the lung. As several reports of anticancer activity of bromelain are available, we have combined EOLE with bromelain to study their protective role against BaP-mediated lung damage. Changes in DNA integrity, LPO level in lung after EOLE-treated animal were examined. Then, we have evaluated the synergistic role of EOLE and bromelain. We have found that EOLE in combination with bromelain was able to increase the translocation of Nrf2 from cytoplasm to nucleus and decrease the translocation of NF-κB from cytoplasm to nucleus. Combination of treatment also reduced the expression of TNFα, IL-6, and some matrix metalloproteinases in lung tissue. Our findings suggest that EOLE and bromelain can synergistically reduce the BaP-induced lung carcinogenesis associated with inflammation and oxidative stress via regulating the expression of various inflammatory markers and also modulating the activity of pulmonary antioxidant armories.

Chemoprotective effect of atorvastatin against benzo(a)pyrene-induced lung cancer via the inhibition of oxidative stress and inflammatory parameters.

BackgroundLung cancer affects approximately 9% of women and 17% of men worldwide, and has a mortality rate of 17%. Previously published studies have suggested that oxidative stress expansion can lead to lung cancer. The aim of the current study was to analyze the possible inhibitory pathway of atorvastatin against lung cancer cells in an in vivo model.MethodsThe cytotoxic effects of atorvastatin on lung cancer cell lines H460 and A549 were analyzed, as well as cell cycle arrest and cell morphology. Benzo(a)pyrene (BaP) was used for the induction of lung cancer in experimental rats, and atorvastatin (5, 10, and 20 mg/kg body weight) was used for treatment in a dose-dependent manner. Body weight and lung tumors were calculated at regular intervals. Antioxidants, pro-inflammatory cytokines, phase I and II antioxidant enzymes, polyamine enzymes, and apoptosis markers were determined at end of the experimental study.ResultsCell cycle arrest occurred at the G2/M phase after atorvastatin treatment. Atorvastatin increased cytochrome C expression and caspase activity in a dose-dependent manner, and increased the activity of antioxidative enzymes, such as GPx, SOD, GST, reduced glutathione, and catalase, and reduced the level of nitrate and LPO. It also altered the xanthine oxidase (XO), Lactic Acid Dehydrogenase (LDH), quinone reductase (QR), UDP-glucuronosyltransferase (UDP-GT), adenosine deaminase (ADA), Aryl hydrocarbon hydroxylase (AHH), 5'-nucleotidase, cytochrome P450, cytochrome B5 and NADPH cytochrome C reductase levels. Atorvastatin was found to modulate polyamine enzyme levels, such as histamine, spermine, spermidine, and putrescine, and significantly (P<0.001) reduced the pro-inflammatory cytokine levels, such as tumor necrosis factor-α. Interleukin (IL)-6 and interleukin-1β (IL-1β) increased caspase-3 and caspase-9 levels in a dose-dependent manner.ConclusionsOur findings indicate that atorvastatin can inhibit lung cancer through apoptosis.

Immunomodulatory effect of d-carvone in swiss albino mice with benzo(a)pyrene-induced lung cancer

Background: Globally, lung cancer is the second most cause of deaths, which accounts for approximately 29% of the cancer cases worldwide. d-Carvone is considered a vital constituent of many essential oils with immense pharmacological benefits. Objective: In this study, we examined the immunomodulatory effect of d-carvone in the Swiss albino mice model against benzo(a)pyrene (BaP)-induced lung carcinoma. Materials and Methods: BaP was orally administered to the mice (50 mg/kg body weight [bw] for 4 weeks [twice a week]). Post-BaP administration, d-carvone (20 mg/kg bw) was orally administered to the mice (20 mg/kg bw). We calculated the tumor incidence and performed the following measurements: lung and bw, hematological parameters, immune complexes (phagocytic and avidity indexes, nitroblue tetrazolium reduction, soluble immune complex levels, immunocompetent cells (leukocytes, lymphocytes, and neutrophils), immunoglobin (Ig) levels (IgG, IgA, and IgM), level of xenobiotics and enzymes that point toward hepatic dysfunction, carcinoembryonic antigen (CEA), and proinflammatory cytokines (PICs) in experimental and normal mice. The level of oxidative stress in the experimental animals was investigated. The lung tissues of investigational animals were examined for the histopathological changes. Results: According to the results, there was an increased level of lipid peroxidation and decreased level of antioxidant activity in the lung tumor samples. The counts of lymphocytes, polymorphonuclear cells, and macrophages were notably decreased and increased, respectively, by the d-carvone post-treatment. Moreover, BaP-induced inflammation that is indicated by the increased in the level of PICs and CEA proteins in lung samples. d-Carvone attenuated the levels of PICs and CEA. Conclusion: The results of this study reveal that d-carvone prevented the cells against BaP-induced inflammation in lung cancer without causing adverse effects.

The cytotoxic effect of Synadenium grantii extract against human lung carcinoma A549 cells and its role in improvement of histopathological and biomarkers changes in Benzo(a)pyrene-induced lung cancer in rats

Lung cancer is one of the most lethal cancers which is causing up to 3 million deaths annually worldwide. Therefore, management of lung cancer needs searching for new chemopreventive agents. This work was designed to inspect the chemopreventive potential of different extracts prepared from branches and leaves of Synadenium grantii for screening their effects on lung cancer cells (A549), then the most active extract was used for combating lung cancer induced in animal model. The in vitro results showed that, the methanolic extract was the most active extract against A549 cells with a notable cytotoxicity activity (IC50: 4.30±0.44 µg/ml), which was close to the activity of standard drug, doxorubicin (IC50: 3.50±0.40 µg/ml). The results of the in vivo experiment, revealed that in B(a)P-treated group, aspartate (AST) and alanine (ALT) transaminase activities as well as the levels of urea, creatinine, alpha-fetoprotein (AFP) and Phosphotylinosital 3 Kinase (PI3K) were significantly increased comparing to control group. However, treatment with S. grantii ameliorated the increase in these parameters in both after- and before-treatment groups comparing with B(a)P-treated group. This improvement in biochemical results were also supported by improving in morphological and histopathological injuries induced by B(a)P, which indicated that methanolic extract of S. grantii has a chemoprevention effect on lung cancer.

IL-27 along with IL-28B ameliorates the pulmonary redox impairment, inflammation and immunosuppression in benzo(a)pyrene induced lung cancer bearing mice

AimsThe major cause behind lung cancer development is exposure to various polycyclic aromatic hydrocarbons like benzo(a)pyrene (BaP) present in tobacco smoke, motor vehicle, and industrial exhaust. BaP is reported to induce the expression of various pro-inflammatory cytokines and matrix remodeling proteins. It is also responsible for dysfunction and exhaustion of the killing capacity of CD8+ T lymphocytes, one of the important components of the immune system which can kill tumor cells. We tried to evaluate the synergistic role of IL-27 and IL-28B in modulation of BaP-induced lung carcinogenesis associated with various hallmarks like pulmonary redox imbalance, angiogenesis, inflammation and cell proliferation in lung tissue. Main methodBaP was treated to Swiss albino mice to develop lung tumor. After the confirmation of lung tumor development Swiss albino mice were treated with IL-27 and IL-28B alone or in combination intraperitoneally. Histological analysis, immunohistochemistry, biochemical assay, western blot analysis, cell cytotoxicity assay, real-time PCR assay etc. were performed to evaluate the modulatory role of IL-27 and IL-28B. Key findingsWe observed that IL-27 and IL-28B were able to suppress the expression of lung cancer-associated NFkB, COX-2, and iNOS. The expression of TNF-α, PCNA and some matrix remodeling enzymes were also modulated upon IL-27 and IL-28B treatment. Although the population of lung residing CD8+ T cells in tumor bearing lung tissue were unresponsive but the activity of systemic CD8+ cells was increased. SignificanceResults hinted that IL-27 along with IL-28B were able to ameliorate various hallmarks ranging from angiogenesis to inflammation associated with the BaP-induced lung carcinogenesis. From this study, we propose that IL-27 and IL28B can be used as immunotherapeutic agent to regulate lung cancer.

Antineoplastic potential of eupatilin against benzo[a]pyrene-induced lung carcinogenesis

Background: Most of the conservative therapies used to treat lung cancer show serious side effects. In addition, the prevalence and death rates due to lung cancer have been increasing alarmingly across the globe. Eupatilin (EUP) is a naturally occurring flavone which is primarily the active ingredient of the traditional Chinese medicine Artemisia asiatica. Materials and Methods: In this study, we evaluated the antineoplastic effect of EUP against benzo(a)pyrene-induced lung cancer in Swiss albino mice. We analyzed the level of xenobiotics, liver dysfunction enzymes (LDEs), pro-inflammatory cytokines, and histology of the liver. Furthermore, we conducted in vitro experiments (A549 cells) to elucidate the amount of cell proliferation, apoptosis, and their markers (caspases 3 and 9). Results: The EUP (30 mg/kg bw) treatment of tumor-bearing mice with EUP revealed the normal levels of xenobiotic, LDEs, antioxidant enzymes, lipid peroxidation in the liver and further carcinoembryonic antigen, pro-inflammatory marker, and histology in lung tissues. EUP inhibited the proliferation of A549 cells and induced the formation of reactive oxygen species and apoptosis by upregulating the expression of caspases 3 and 9. Conclusion: Overall, these results substantiate the anti-neoplastic effects of EUP against carcinogen-induced lung cancer in in vitro and in vivo models.

Protective effect of vanillic acid against benzo(a)pyrene induced lung cancer in Swiss albino mice.

Vanillic acid (VA) is found in high concentrations in various plants and used as traditional medicinefor various diseases. The aim of the existing study is to illustrate the protective effects of VA against benzo(a)pyrene (B(a)P)-induced lung cancer in Swiss albino mice. B(a)P (50 mg/kg b.wt.) was given orally to induce lung cancer in mice. The body weight, tumor incidence, carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and enzymatic/nonenzymatic antioxidants (superoxide dismutase, catalase, glutathione reductase, glutathione peroxidase, and glutathione) were estimated. Further histochemical investigation through hematoxylin and eosin staining was also carried out. B(a)P administered groups showed increased levels of serum pathological markers CEA,NSE along with reduced final body weight as well as decreased tissue enzymatic and nonenzymatic antioxidants activities, whereas VA treatment (200mg/kg/b.wt) along with B(a)P showed significantly reverted the above changes, which proves as prominent anticancer effects in experimentally induced lung cancer. Overall, these results suggest that VA has an efficient preventive action against B(a)P-induced lung cancer, and this is attributed to its free-radical scavenging antioxidant activities.

Anti-Inflammatory and Apoptotic Signaling Effect of Fucoxanthin on Benzo(A)Pyrene-Induced Lung Cancer in Mice.

Fucoxanthin, a potent carotenoid present in various natural sources especially from seaweeds; it exhibits several biological effects like anti-neoplastic, anti-mutagenic, anti-diabetic, anti-obesity and anti-inflammatory actions. Fucoxanthin role in chemoprevention of lung cancer in mouse model induced using benzo(a)pyrene [B(a)P] has been presented here. Oral administration of fucoxanthin with and without B(a)P were studied, the results from our study shows that fucoxanthin significantly decreased tumor progression in mice exposed to B(a)P, the obtained data were correlated with increased antioxidant, apoptosis and decreased tumour marker and anti-apoptotic molecules. With respect to apoptosis, fucoxanthin treated animals shows increased apoptosis compared to tumor induced mice by increased expression of caspase 9 and 3 and decreased expression of anti-apoptotic Bcl2 protein. Finally, histopathological and immuno histochemical analysis also revealed that fucoxanthin shows potent anticancer agent by bringing back the damaged tissue treated with B(a)P and also decreases the expression of PCNA in cancer induced mice. The anticancer effect of fucoxanthin may be attributed by several independent mechanisms which play a important roles in the prevention of cancer development, there is also substantial evidences to show that fucoxanthin acts indirectly by increasing the antioxidant capacity of affected tissue and prepared to cope up with oxidative stress which is proved in our study. Thus from our study it is clearly established that fucoxanthin act as a persuasive anticancer drug against lung cancer.

Enhancement of tumor suppressor RAR-β protein expression by cationic liposomal-ATRA treatment in benzo(a)pyrene-induced lung cancer mice model.

Targeting the specific molecular proteins or genes which are responsible for the suppression of cancer growth is currently an emerging molecular method to treat cancer. ATRA treatment is now considered as a molecular targeted therapy for many cancers. As ATRA exhibits its therapeutic effect through its receptors, this study was focused to investigate the effect and action of liposomal-ATRA treatment on the expression of RAR-β protein which is also a tumor suppressor. The liposomal-ATRA was developed with cationic DOTAP and cholesterol by thin-film formation method. The benzo(a)pyrene(50mg/kg b.wt)-induced mice were treated with free and liposomal-ATRA(0.60mg/kg b.wt). The RAR-β protein expression in lung and liver tissue samples were analyzed by immunohistochemistry (IHC) and western blotting (WB) on the 30th and 120th days. Almost nil expression of RAR-β protein was observed in B(a)P cancer control and liposome alone-treated groups. A comparatively elevated expression was seen in the free ATRA-treated group (IHC score-2+ in lung on the 120th day with band density of 14.46 ± 1.24% in WB). Interestingly, the liposomal-ATRA treatment demonstrated a significantly (p ≤ 0.01) higher RAR-β expression in lung (35.20 ± 3.398% band intensity and score 4+ in the 120th day) than that of in ATRA alone treatment. This study results indicate that the RAR-β protein expression was suppressed by B(a)P during cancer induction even on the 30th day itself. The treatment could reactivate the suppression and the lipo-ATRA treatment could show significantly higher RAR-β expression on the 120th day, which implies that it accumulated more ATRA in target site and sustained it for enhanced action.

Reduced RAR-β gene expression in Benzo(a)Pyrene induced lung cancer mice is upregulated by DOTAP lipo-ATRA treatment

Molecular targeted therapy for specific genes is an emerging research. Retinoic Acid Receptor (RAR-β) is a key tumor suppressor which is found to be lost drastically during much cancer progression. We hence, analyzed the expression level of RAR-β gene during B(a)P induced lung cancer development in mice and studied the lung cancer targeted action of All Trans Retinoic Acid (ATRA) in DOTAP liposomal formulation. The effect of its treatment on lung cancer was determined by histopathological analysis. RAR-β gene expression was assessed by RT-PCR and qPCR. A distinct band for RAR-β gene (density - 0.5123 for lung and 0.5160 for liver) was observed in normal mice, whereas no visible band was observed in cancer induced group, indicating loss of RAR-β gene expression. Both ATRA and lipo-ATRA treated groups showed detectable RAR-β expression with relatively lesser density than the normal group. The expression was more intense in lipo-ATRA treatment (density-0.2973) compared with free ATRA treatment (density-0.1549) in lung tissues. The qPCR results also have highlighted a highly significant (p ≤ 0.01) variation RQ values between lipo-ATRA group (15.46 ± 1.54) and free ATRA group (7.58 ± 1.30) in lung tissue sample on 30th day. The mean RQ value for normal lung on 30th day was 20.86 ± 2.58 against the cancer control. The 120th day mice also showed the similar RAR-β expression pattern with further declined expression levels as there was no treatment given after 30 days. Interestingly, the lipo-ATRA treatment could show a highly significant (p ≤ 0.001) expression (12.00 ± 2.31) when compared with free ATRA treatment (3.31 ± 0.58) which implies that the lipo-ATRA formulation could result in sustained delivery of ATRA in target site. Histopathology of lung and liver on 120th day also revealed an effective therapeutic indication in lipo-ATRA treatment compared to free ATRA treatment due to lipo-ATRA's stealth property and it efficiently inhibited the metastasis to liver. These results revealed that the lipo-ATRA treatment has efficiently delivered ATRA into target site than free ATRA and in-turn it might have induced the expression of RAR-β gene or prevented loss of RAR-β gene in cancer animals.

Combination of carboplatin and intermittent normobaric hyperoxia synergistically suppresses benzo[a]pyrene-induced lung cancer.

Background/AimsWe explored the effects of intermittent normobaric hyperoxia alone or combined with chemotherapy on the growth, general morphology, oxidative stress, and apoptosis of benzo[a]pyrene (B[a]P)-induced lung tumors in mice.MethodsFemale A/J mice were given a single dose of B[a]P and randomized into four groups: control, carboplatin (50 mg/kg intraperitoneally), hyperoxia (95% fraction of inspired oxygen), and carboplatin and hyperoxia. Normobaric hyperoxia (95%) was applied for 3 hours each day from weeks 21 to 28. Tumor load was determined as the average total tumor numbers and volumes. Several markers of oxidative stress and apoptosis were evaluated.ResultsIntermittent normobaric hyperoxia combined with chemotherapy reduced the tumor number by 59% and the load by 72% compared with the control B[a]P group. Intermittent normobaric hyperoxia, either alone or combined with chemotherapy, decreased the levels of superoxide dismutase and glutathione and increased the levels of catalase and 8-hydroxydeoxyguanosine. The Bax/Bcl-2 mRNA ratio, caspase 3 level, and number of transferase-mediated dUTP nick end-labeling positive cells increased following treatment with hyperoxia with or without chemotherapy.ConclusionsIntermittent normobaric hyperoxia was found to be tumoricidal and thus may serve as an adjuvant therapy for lung cancer. Oxidative stress and its effects on DNA are increased following exposure to hyperoxia and even more with chemotherapy, and this may lead to apoptosis of lung tumors.

P1.03-025 Combination Therapy with Carboplatin and Hyperoxia Synergistically Enhances Suppression of Benzo[a]Pyrene Induced Lung Cancer

P1.03-025 Combination Therapy with Carboplatin and Hyperoxia Synergistically Enhances Suppression of Benzo[a]Pyrene Induced Lung Cancer

Ferulenol, a Sesquiterpene Coumarin, Induce Apoptosis via Mitochondrial Dysregulation in Lung Cancer Induced by Benzo[a]pyrene: Involvement of Bcl2 Protein.

Ferulenol, a sesquiterpene coumarin, was extracted from "Ferula vesceritensis" and possesses pro-oxidative and anticancer effects in different types of cancer. The objective of this study is to determine whether ferulenol has an anticancer effect by regulation the Bcl2 protein expression in lung cancer induced by benzo[a]pyrene in Wistar rats. Rats in group 1 have received, intraperitoneally, olive oil and considered as controls, animals of group 2 were treated with 100 mg/kg of benzo[a]pyrene intraperitoneally in order to induce lung cancer for 24 weeks, the 3rd groups of rats received the ferulenol 50 mg/kg intraperitoneally after 24 weeks of administration of benzo[a]pyrene and the last group, the rats were treated with ferulenol alone 50 mg/kg. Treatment with ferulenol significantly increased the rate of lipid peroxidation and decrease enzymatic (CAT and GST) and non-enzymatic (GSH) anti-oxidants in benzo[a]pyrene induced lung cancer. Anticancer efficacy of ferulenol was evaluated by down-regulation of Bcl2 protein and up-regulation of Bax protein, Therefore, ferulenol can alter the functionality of lung mitochondria by increasing the production of superoxide anion and mitochondrial swelling. Together, our results depict that ferulenol can be used as pro-oxidant and chemotherapeutic agent against lung cancer.