Animal Models Of Lung Cancer Research Articles

Rho GTPases RhoA and Rac1 Mediate Effects of Dietary Folate on Metastatic Potential of A549 Cancer Cells through the Control of Cofilin Phosphorylation

Folate, an important nutrient in the human diet, has been implicated in cancer, but its role in metastasis is not established. We have shown previously that the withdrawal of medium folate leads to the inhibition of migration and invasion of A549 lung carcinoma cells. Here we have demonstrated that medium folate regulates the function of Rho GTPases by enabling their carboxyl methylation and translocation to plasma membrane. Conversely, the lack of folate leads to the retention of these proteins in endoplasmic reticulum. Folate also promoted the switch from inactive (GDP-bound) to active (GTP-bound) GTPases, resulting in the activation of downstream kinases p21-activated kinase and LIM kinase and phosphorylation of the actin-depolymerizing factor cofilin. We have further demonstrated that in A549 cells two GTPases, RhoA and Rac1, but not Cdc42, are immediate sensors of folate status: the siRNA silencing of RhoA or Rac1 blocked effects of folate on cofilin phosphorylation and cellular migration and invasion. The finding that folate modulates metastatic potential of cancer cells was confirmed in an animal model of lung cancer using tail vein injection of A549 cells in SCID mice. A folate-rich diet enhanced lung colonization and distant metastasis to lymph nodes and decreased overall survival (35 versus 63 days for mice on a folate-restricted diet). High folate also promoted epithelial-mesenchymal transition in cancer cells and experimental mouse tumors. Our study provides experimental evidence for a mechanism of metastasis promotion by dietary folate and highlights the interaction between nutrients and metastasis-related signaling.

DNMT3B Overexpression by Deregulation of FOXO3a-Mediated Transcription Repression and MDM2 Overexpression in Lung Cancer

DNA methyltransferase 3B (DNMT3B) contributes to de novo DNA methylation and its overexpression promotes tumorigenesis. However, whether DNMT3B is upregulated by transcriptional deregulation remains unclear. We studied the transcriptional repression of DNMT3B by forkhead O transcription factor 3a (FOXO3a) in lung cancer cell, animal, and clinical models. The results of luciferase reporter assay showed that FOXO3a negatively regulated DNMT3B promoter activity by preferentially interacting with the binding element FOXO3a-E (+166 to +173) of DNMT3B promoter. Ectopically overexpressed FOXO3a or combined treatment with doxorubicin to induce FOXO3a nuclear accumulation further bound at the distal site, FOXO3a-P (-249 to -242) by chromatin-immunoprecipitation assay. Knockdown of FOXO3a resulted in an open chromatin structure and high DNMT3B mRNA and protein expression. Abundant FOXO3a repressed DNMT3B promoter by establishing a repressed chromatin structure. Note that FOXO3a is a degradation substrate of MDM2 E3-ligase. Cotreatment with doxorubicin and MDM2 inhibitor, Nutlin-3, further enforced abundant nuclear accumulation of FOXO3a resulting in decrease expression of DNMT3B leading to synergistic inhibition of tumor growth and decrease of methylation status on tumor suppressor genes in xenograft specimens. Clinically, lung cancer patients with DNMT3B high, FOXO3a low, and MDM2 high expression profile correlated with poor prognosis examined by immunohistochemistry and Kaplan-Meier survival analysis. We reveal a new mechanism that FOXO3a transcriptionally represses DNMT3B expression and this regulation can be attenuated by MDM2 overexpression in human lung cancer model. Cotreatment with doxorubicin and Nutlin-3 is a novel therapeutic strategy through epigenetic modulation.

Alternative animal model of urethane-induce lung cancer: A pilot study

Alternative animal model of urethane-induce lung cancer: A pilot study

Treatment Guidance from the Blood

Circulating tumor cells from small-cell lung cancer yield genomic information and animal models.

SU-E-T-217: Intrinsic Respiratory Gating in Small Animal CT

Purpose: Preclinical animal models of lung cancer can provide a controlled test-bed for testing dose escalation or function-based-treatment-planning studies. However, to extract lung function, i.e. ventilation, one needs to be able to image the lung at different phases of ventilation (in-hale / ex-hale). Most respiratory-gated imaging using micro-CT involves using an external ventilator and surgical intervention limiting the utility in longitudinal studies. A new intrinsic respiratory retrospective gating method was developed and tested in mice. Methods: A fixed region of interest (ROI) that covers the diaphragm was selected on all projection images to estimate the mean intensity (M). The mean intensity depends on the projection angle and diaphragm position. A 3-point moving average (A) of consecutive M values: Mpre, Mcurrent and Mpost, was calculated to be subtracted from Mcurrent. A fixed threshold was used to enable amplitude based sorting into 4 different phases of respiration. Images at full-inhale and end-exhale phases of respiration were reconstructed using the open source OSCaR. Lung volumes estimated at the 2 phases of respiration were validated against literature values. Results: Intrinsic retrospective gating was accomplished without the use of any external breathing waveform. While projection images were acquired at 360 different angles. Only 138 and 104 projections were used to reconstruct images at full-inhale and end-exhale. This often results in non-uniform under-sampled angular projections leading to some minor streaking artifacts. The calculated expiratory, inspiratory and tidal lung volumes correlated well with the values known from the literature. Conclusion: Our initial result demonstrates an intrinsic gating method that is suitable for flat panel cone beam small animal CT systems. Reduction in streaking artifacts can be accomplished by oversampling the data or using iterative reconstruction methods. This initial experience will enable freebreathing small animal micro-CT imaging to fuel longitudinal studies of lung function.

Oleanane triterpenoids in the prevention and therapy of breast cancer: current evidence and future perspectives.

Breast cancer is one of the most frequently diagnosed cancers and major cause of death in women in the world. Emerging evidence underscores the value of dietary and non-dietary phytochemicals, including triterpenoids, in the prevention and treatment of breast cancer. Oleanolic acid, an oleanane-type pentacyclic triterpenoid, is present in a large number of dietary and medicinal plants. Oleanolic acid and its derivatives exhibit several promising pharmacological activities, including antioxidant, anti-inflammatory, hepatoprotective, cardioprotective, antipruritic, spasmolytic, antiallergic, antimicrobial and antiviral effects. Numerous studies indicate that oleanolic acid and other oleanane triterpenoids modulate multiple intracellular signaling pathways and exert chemopreventive and antitumor activities in various in vitro and in vivo model systems. A series of novel synthetic oleanane triterpenoids have been prepared by chemical modifications of oleanolic acid and some of these compounds are considered to be the most potent anti-inflammatory and anticarcinogenic triterpenoids. Accumulating studies provide extensive evidence that synthetic oleanane derivatives inhibit proliferation and induce apoptosis of various cancer cells in vitro and demonstrate cancer preventive or antitumor efficacy in animal models of blood, breast, colon, connective tissue, liver, lung, pancreas, prostate and skin cancer. This review critically examines the potential role of oleanolic acid, oleanane triterpenoids and related synthetic compounds in the chemoprevention and treatment of mammary neoplasia. Both in vitro and in vivo studies on these agents and related molecular mechanisms are presented. Several challenges and future directions of research to translate already available impressive preclinical knowledge to clinical practice of breast cancer prevention and therapy are also presented.

Inhibition of human lung cancer cell proliferation and survival by wine

BackgroundCompounds of plant origin and food components have attracted scientific attention for use as agents for cancer prevention and treatment. Wine contains polyphenols that were shown to have anti-cancer and other health benefits. The survival pathways of Akt and extracellular signal-regulated kinase (Erk), and the tumor suppressor p53 are key modulators of cancer cell growth and survival. In this study, we examined the effects of wine on proliferation and survival of human Non-small cell lung cancer (NSCLC) cells and its effects on signaling events.MethodsHuman NSCLC adenocarcinoma A549 and H1299 cells were used. Cell proliferation was assessed by thymidine incorporation. Clonogenic assays were used to assess cell survival. Immunoblotting was used to examine total and phosphorylated levels of Akt, Erk and p53.ResultsIn A549 cells red wine inhibited cell proliferation and reduced clonogenic survival at doses as low as 0.02%. Red wine significantly reduced basal and EGF-stimulated Akt and Erk phosphorylation while it increased the levels of total and phosphorylated p53 (Ser15). Control experiments indicated that the anti-proliferative effects of wine were not mediated by the associated contents of ethanol or the polyphenol resveratrol and were independent of glucose transport into cancer cells. White wine also inhibited clonogenic survival, albeit at a higher doses (0.5-2%), and reduced Akt phosphorylation. The effects of both red and white wine on Akt phosphorylation were also verified in H1299 cells.ConclusionsRed wine inhibits proliferation of lung cancer cells and blocks clonogenic survival at low concentrations. This is associated with inhibition of basal and EGF-stimulated Akt and Erk signals and enhancement of total and phosphorylated levels of p53. White wine mediates similar effects albeit at higher concentrations. Our data suggest that wine may have considerable anti-tumour and chemoprevention properties in lung cancer and deserves further systematic investigation in animal models of lung cancer.

Abstract B08: Triptolide has antitumor effects in lung cancer via Wnt pathway inhibition.

Abstract Introduction: Triptolide is an extract of the herb Tripterygium wilfordii (Tw), which has been used to treat autoimmune disorders in traditional Chinese medicine. It has also been reported to have anti-cancer effects in vitro, although little is known about its mechanism of action. The Wnt/B-catenin pathway has been implicated in regulating cancer stem cells in lung cancer, making this pathway a potential target for novel lung cancer therapies. Methods: Here we tested the efficacy of triptolide in several lung cancer cell lines. We elucidated a Wnt pathway mechanism for triptolide effect in lung cancer, first by using a Wnt focused cDNA array in vitro. We examined the effect of triptolide on tumor cell death using flow cytomotery and a beta-galactosidase assay. We tested the ability of triptolide to prevent development of lung tumors in the E160D transgenic mouse model of lung cancer, which carries a somatic mutation in the DNA repair enzyme flap endonuclease-1 (FEN-1). Twelve month old mice were fed triptolide 1mg/kg daily for 2 months and were sacrificed at 18 months along with a control group. Results: Triptolide had more potent therapeutic efficacy in multiple lung cancer cell lines compared with cisplatin and paclitaxel, in vitro. In H460 and A549 cell lines, we found that triptolide resulted in Wnt pathway silencing via overexpression of Wnt inhibitory factor-1 (WIF-1), which occurs due to promoter demethylation. Addition of recombinant Wnt3a, which causes Wnt pathway activation, partially rescues the effect of triptolide on lung cancer cell lines. Triptolide resulted in cellular senescence as demonstrated by beta-galactosidase assay. Finally, in the E160D transgenic mouse model of lung cancer, a two month course of oral triptolide treatment decreased the development of lung tumors from 70% to 30% at 18 months (p=0.01). There were no observed clinical toxicities at the dosage of triptolide used. Conclusions: Triptolide has potent anti-cancer effects in vitro in lung cancer cell lines, and significantly decreases lung tumor development in a transgenic mouse model. Triptolide suppresses the Wnt pathway by causing upregulation of WIF-1a by promoter demethylation. Future studies are needed to test this promising and novel Wnt-targeting therapy in lung cancer animal models, and to identify biomarkers which may predict sensitivity in particular subgroups of lung cancer patients. Citation Format: Youqiang Li, Li Zheng, Binghui Shen, Jae Kim, Dan Raz. Triptolide has antitumor effects in lung cancer via Wnt pathway inhibition. [abstract]. In: Proceedings of the AACR-IASLC Joint Conference on Molecular Origins of Lung Cancer; 2014 Jan 6-9; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2014;20(2Suppl):Abstract nr B08.

Animal models of lung cancer

In order to make a rational and effective cure measure for lung cancer,people need to establish suitable animal model.Spontaneous or induced model needs long time to form tumor and the rate is not steady.Xenograft mouse model is widely used in laboratories because of its steady tumor formation rate and less time in oncogenesis.Engineered mouse model which is based on genetic technology not only helps us to understand pathological process of lung cancer,but also can provide an ideal preclinical model of targeted therapy experiment,and it will be the important development direction of lung cancer animal model. Key words: Lung neoplasms; Models, animal

Cytosolic PhospholipaseA2 Inhibition with PLA-695 Radiosensitizes Tumors in Lung Cancer Animal Models

Lung cancer remains the leading cause of cancer deaths in the United States and the rest of the world. The advent of molecularly directed therapies holds promise for improvement in therapeutic efficacy. Cytosolic phospholipase A2 (cPLA2) is associated with tumor progression and radioresistance in mouse tumor models. Utilizing the cPLA2 specific inhibitor PLA-695, we determined if cPLA2 inhibition radiosensitizes non small cell lung cancer (NSCLC) cells and tumors. Treatment with PLA-695 attenuated radiation induced increases of phospho-ERK and phospho-Akt in endothelial cells. NSCLC cells (LLC and A549) co-cultured with endothelial cells (bEND3 and HUVEC) and pre-treated with PLA-695 showed radiosensitization. PLA-695 in combination with irradiation (IR) significantly reduced migration and proliferation in endothelial cells (HUVEC & bEND3) and induced cell death and attenuated invasion by tumor cells (LLC &A549). In a heterotopic tumor model, the combination of PLA-695 and radiation delayed growth in both LLC and A549 tumors. LLC and A549 tumors treated with a combination of PLA-695 and radiation displayed reduced tumor vasculature. In a dorsal skin fold model of LLC tumors, inhibition of cPLA2 in combination with radiation led to enhanced destruction of tumor blood vessels. The anti-angiogenic effects of PLA-695 and its enhancement of the efficacy of radiotherapy in mouse models of NSCLC suggest that clinical trials for its capacity to improve radiotherapy outcomes are warranted.

Combining bevacizumab with endostatin gets better antitumor efficacy in vivo in lung cancer animal model

背景与目的研究重组人血管内皮抑制素和贝伐珠单抗在体内对肺腺癌抑制作用的差别及联合用药的效果。方法首先建立A549肺腺癌细胞系的荷瘤Balb/c小鼠动物模型，然后将小鼠随机分为4组，对照组使用普通生理盐水每日瘤周注射。重组人血管内皮抑制素治疗组使用重组人血管内皮抑素（3 mg/kg）每日瘤周注射连续16天贝伐珠单抗治疗组使用贝伐珠单抗（5 mg/kg）每周两次瘤周注射给药。贝伐珠单抗、重组人血管内皮抑制素联合用药组使用贝伐珠单抗（5 mg/kg）每周两次瘤周注射给药+重组人血管内皮抑素（3 mg/kg）每日瘤周注射给药。治疗16天后处死所有实验鼠切取肿瘤标本比较实体瘤大小，采用Western blot的方法检测血管内皮生长因子A和C（vascular endothelial growth factor/VEGF-A, C）在各组表达情况的差异。结果重组人血管内皮抑制素和贝伐珠单抗在体内实验中均表现出了抑制肿瘤生长的作用，贝伐珠单抗作用更加明显（52.36% vs 38.68%）。联合使用可获得更好的效果（64.15%）。贝伐珠单抗只对VEGF-A有抑制作用（60.8%），重组人血管内皮抑制素对VEGF-A/C都有抑制作用（14.6%, 30.3%）。联合用药组对VEGF-A/C的抑制作用最强（79.4%, 44.2%）。结论重组人血管内皮抑制素和贝伐珠单抗都在裸鼠动物模型试验中表现出了明显的抑瘤效果，联合使用抑制肿瘤效果更加明显。恩度对肿瘤组织内VEGF-A/C都表现出了抑制作用贝伐珠单抗对VEGF-A表现出了较明显的抑制作用联合用药组对VEGF-A/C抑制作用更加明显。

Dietary phosphate: what do we know about its toxicity

Inorganic phosphate (Pi) is an essential mineral required for diverse cellular processes. Recent genetic and dietary experiments in animal models indicate that Pi may be toxic to a variety of biological processes. High dietary Pi load in such animal models resulted in an increase in oxidative stress, DNA damage that resulted in phenotypic expression of premature aging, and short life span. Further, high Pi load was reported to induce carcinogenesis in lung and skin cancer animal models. So far, translational research is limited to observational studies that show an independent association of Pi intake with morbidity and mortality across all strata of kidney function. Fast-food and processed-food consumption in an average American diet leads to large increases in daily phosphate intake. Whether such dietary patterns explain the epidemiology of disease processes in humans remains to be further investigated. Phosphotoxicity is a novel concept with a potential for a large public health impact in future, but urgent studies are needed to prove reducing Pi intake to a certain target generates better clinical outcomes.

An Experimental Model for Induction of Lung Cancer in Rats by Chlamydia Pneumoniae

To assess induction effects of Chlamydia pneumoniae (Cpn) on lung cancer in rats. A lung cancer animal model was developed through repeated intratracheal injection of Cpn (TW-183) into the lungs of rats, with or without exposure to benzo(a)pyrene (Bp). Cpn antibodies (Cpn-IgA, -IgG, and -IgM) in serum were measured by microimmunofluorescence. Cpn-DNA or Cpn-Ag of rat lung cancer was detected through polymerase chain reaction or enzyme-linked immunosorbent assay. The prevalence of Cpn infection was 72.9% (35/48) in the Cpn group and 76.7% (33/43) in the Cpn plus benzo(a)pyrene (Bp) group, with incidences of lung carcinomas in the two groups of 14.6% (7/48) and 44.2% (19/43), respectively (P-values 0.001 and <0.001 compared with normal controls). A rat model of lung carcinoma induced by Cpn infection was successfully established in the laboratory for future studies on the treatment, prevention, and mechanisms of the disease.

Overexpression of PAFAH1B1 is associated with tumor metastasis and poor survival in non-small cell lung cancer

Our previous array-comparative genomic hybridization study showed that PAFAH1B1 gene locus was amplified in lung cancer patients, suggesting that PAFAH1B1 is a potential oncogene in lung cancer. Here, we investigate the oncogenic mechanisms of PAFAH1B1 in lung cancer. PAFAH1B1 was characterized in cell and animal models of lung cancer by in vitro migration and invasion assays and in vivo metastasis studies. The mRNA and protein expression levels of PAFAH1B1 were further determined and the prognostic effects of PAFAH1B1 overexpression in lung cancer patients were analyzed. Overexpression of PAFAH1B1 enhanced migration and invasion in lung cancer cells, whereas knockdown of PAFAH1B1 decreased cell migration and invasion, and disrupted cell microtubule organization and pericellular poly-fibronectin assemblies. In vivo tumor metastasis assay confirmed that PAFAH1B1 knockdown in lung cancer cells markedly reduced their metastasis capabilities in animals. The frequencies of overexpressed PAFAH1B1 mRNA and protein were 62.4% (63/101) and 57.4% (58/101) in lung cancer patients, respectively. The clinical correlation results showed that overexpression of PAFAH1B1 was significantly associated with late stage (mRNA: P=0.008, protein: P=0.008) and poor survival in lung adenocarcinoma (P=0.020) and male patients (P=0.049). Our results provide the first evidence that PAFAH1B1 overexpression contributes to lung tumorigenesis and poor prognosis. These effects are partly mediated through disruption of microtubule network and pericellular poly-fibronectin assembly to promote migration and invasiveness of lung cancer cells.

Abstract 2835: Discovery of novel inhibitors for ECT2 as a novel therapeutic strategy for lung cancer

Abstract The Epithelial Cell Transforming sequence 2 (ECT2) proto-oncogene is a Guanine Exchange Factor (GEF) for RhoA, Rac1 and Cdc42 and is essential to the regulation of cytokinesis. ECT2 contains the Dbl homology and pleckstrin homology (PH) domains, which are the hallmarks of GEFs. ECT2 is over-expressed in primary non-small cell lung cancer (NSCLC) tumors, and injection of ECT2 transfectants into nude mice efficiently induces tumor formation. High level of ECT2 expression is associated with poor prognosis for patients with NSCLC. Knock down of ECT2 expression by small interfering RNAs effectively suppresses lung cancer cell growth, suggesting a specific role of ECT2 in lung cancer development. Taken together, ECT2 may represent an attractive molecular target for inhibiting lung tumor growth. Our studies are based on the hypothesis that ECT2 plays an important role in lung cancer progression and is a novel target for the development of drugs to treat lung cancer. We have built a model for ECT2 PH domain using protein homology modeling. Docking of new compounds selected from commercial drug-like libraries has led to the identification of novel inhibitors of ECT2 PH domain. We have identified several compounds that bind ECT2 PH domain in the low micromolar range (KD&amp;lt;1μM) as measured using surface plasmon resonance spectroscopy and that inhibit the proliferation of a panel of 23 lung cancer cell lines (EC50&amp;lt;50μM). Importantly, inhibition occurred only in mutant-KRAS cells. Two compounds were shown to inhibit actin polymerization using a phalloidin-rhodamine stain as well as in vitro tumor formation using a colony forming assay and migration using a wound healing assay. These two compounds are being evaluated for their anti-tumor activity against several human lung cancer xenografts in SCID mice. The objectives of our studies are 1) to conduct molecular mechanistic studies of the precise role of ECT2 in the development of lung cancer in vivo animal model of lung cancer; 2) to test the most active compounds in cells in culture to provide a mechanistic rationale for their biological activity and 3) to test the anti-tumor properties of the lead compounds in a in vivo model of lung cancer. The overall goals of our work are to validate ECT2 as a therapeutic target for lung cancer and to develop a novel therapeutic strategy and agents for the improved treatment and the eventual cure of lung cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2835. doi:1538-7445.AM2012-2835

Abstract B17: Inhibition of lung tumor progression in a metastatic mouse model following intravenous delivery of siRNA nanocomplexes

Abstract Exploiting the RNAi pathway offers the potential to advance the treatment of many diseases through highly specific and efficient silencing of gene products. Unfortunately, the requisite safe and efficient delivery of nucleic acids to target cells remains a fundamental problem for the development of RNA and DNA based therapeutics. We have developed a versatile lipopolyamine based delivery system that has been optimized for in vivo delivery by incorporating functional groups onto the core cationic lipopolyamine structure. The direct modification of the core structure (named Staramine) allows for the generation of nanoparticle formulations with a wide range of physicochemical properties and does not require the co-formulation of commercial helper or pegylated lipids. We have previously reported safe, efficient and persistent siRNA mediated transcript knockdown specifically in the lung endothelium using the Starmaine delivery system. Here we extend those observations by administering a siRNA that targets vascular endothelial growth factor receptor 2 (VEGFR-2; KDR/Flk-1) in an animal model of metastatic lung cancer. VEGFR-2, a receptor tyrosine kinase, has been shown to be mainly expressed on endothelial cells and plays a critical role in cell proliferation and differentiation and consequently is also important in angiogenesis associated tumor growth and metastasis. Repeated intravenous administration of nanocomplexes comprised of Staramine formulated VEGFR-2 siRNA (~2 mg/kg/dose) resulted in a significant (40%) reduction of VEGFR-2 transcript levels in isolated tumors from the lungs of mice compared to control injected animals. Similar levels of transcript knockdown were not achieved in tumors when using siRNAs against targets not having the high level of endothelial cell restriction that is seen with VEGFR-2. Immunohistopathology of the lungs of tumor bearing mice administered VEGFR-2 siRNA indicated a significant decrease in vascular density in lung tumors and was consistent with an overall reduction in tumor burden in the mouse lungs. These results support the continued evaluation of using Staramine to deliver siRNAs and other similar molecules as potential therapies for diseases of the lung where the vascular endothelium may be involved. Citation Format: Jason G. Fewell, Khursheed Anwer, Kevin Polach, Majed Matar, Jennifer Rice, Angela Rea-Ramsey, Jeff Sparks, Diane McClure, Casey Pence, Elaine Brunhoeber, Leslie Wilkinson. Inhibition of lung tumor progression in a metastatic mouse model following intravenous delivery of siRNA nanocomplexes [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer; 2012 Jan 8-11; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Res 2012;72(2 Suppl):Abstract nr B17.

Cytosolic PhospholipaseA 2 (cPLA2) Inhibition with Girapladib Radiosensitizes Endothelium and is Effective in Non-small Cell Lung Cancer (NSCLC) Animal Models

Cytosolic PhospholipaseA 2 (cPLA2) Inhibition with Girapladib Radiosensitizes Endothelium and is Effective in Non-small Cell Lung Cancer (NSCLC) Animal Models

Effect of magnetic fluid hyperthermia on lung cancer nodules in a murine model

The purpose of the present study was to investigate the therapeutic effect of magnetic fluid hyperthermia (MFH) induced by an alternating magnetic field (AMF) on human carcinoma A549 xenograft in nude mice. An animal model of human lung cancer was established by subcutaneous injection of human lung cancer A549 cells in BALB/c nude mice. The xenograft mice were randomly divided into four groups and each group was treated with an injection of a different concentration of magnetic fluid: control, low-dose (67.5 mg/ml), medium-dose (90.0 mg/ml) and high-dose group (112.5 mg/ml), respectively. Following the injection (24 h), the tumor was heated in an AMF for 30 min. Tumor volumes were then measured every week. The therapeutic effect was assessed by measuring the tumor volume and weight. Pathological examination was performed with a light and electronic microscope following treatment. The temperature at the surface of the tumor in the low-, medium- and high-dose groups increased to 41.3, 44.5 and 46.8°C, respectively. The tumor grew significantly slower in the medium- and high-dose groups (both p<0.05) compared to the control group. Cytoclasis and apoptosis were detected under light and electron microscopy. In conclusion, MFH induced by AMF inhibited tumor growth and promoted apoptosis of human carcinoma A549 cells in a xenograft mice model.

Abstract 3543: Anti-tumor activity of the vascular disruption agent BNC105 in models of mesothelioma and lung cancer

Abstract BNC105 is a small molecule that exerts anti-cancer activity through disruption of tubulin polymerisation. BNC105 is unique in its ability to selectively target activated endothelial cells usually found in tumor blood vessels. A prodrug formulation, BNC105P, is under evaluation in phase II clinical trials for mesothelioma and renal cancer as a 10min IV infusion. In a xenograft model utilizing the human lung cancer cell line Calu-6, BNC105P caused significant tumor vascular disruption following a single dose. Similar levels of vascular disruption were observed in xenograft tumors arising from subcutaneous injection of the human mesothelioma cell line MSTO-211H. In addition, near complete vascular disruption was observed in lung metastatic lesions in a syngeneic kidney orthotopic model using the murine renal cancer cell line RENCA. We conducted preclinical evaluations that provide evidence of the activity of BNC105 in animal models of mesothelioma and lung cancer as a monotherapy and in combination with current standard of care therapies. Tumor doubling time, tumor growth delay and animal survival were assessed. Animal survival end points were defined in terms of tumor volume, with animals bearing tumors &amp;gt;2000mm3 being euthanised. Cisplatin and Gemcitabine are currently used in first line treatment of non small cell lung cancer. The effect of BNC105P treatment as a monotherapy or in combination with these two agents was investigated in a xenograft model of lung cancer. Mice carrying subcutaneous Calu-6 tumors were treated with two BNC105P doses 1 week apart as a monotherapy or in combination with Cisplatin or Gemcitabine. Combined therapy with BNC105P and Cisplatin resulted in reduced tumor growth. Tumor doubling time for the combination was 27 days, whereas the doubling time for treatment with BNC105P or Cisplatin monotherapies was 20 days and 10 days respectively. The tumor growth delay seen with the combination was 17 days greater compared to cisplatin alone with all animals in the combination arm showing tumor regression during this period. Significant increases in survival were also observed with 100% of animals in the combination group surviving, compared to 33% in the BNC105P and 0% in the Cisplatin treatment groups. In combination with Gemcitabine, BNC105P conferred tumor growth delay that was 3 days greater than BNC105P monotherapy and 17 days greater than Gemcitabine monotherapy. The survival rate in the combination group was 55% compared to 33 % in BNC105P monotherapy group and 0% in the Gemcitabine only group. Pemetrexed in combination with Cisplatin is the currently approved first line treatment for mesothelioma. The therapeutic potential of BNC105P was compared to Cisplatin and Pemetrexed in a xenograft model of the human mesothelioma cell line MSTO-211H. Compared to control, BNC105P increased survival rate by 40% with Cisplatin and Pemetrexed monotherapies increasing survival rates by 20%. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 3543. doi:10.1158/1538-7445.AM2011-3543

Zoledronic acid induces cell-cycle prolongation in murine lung cancer cells by perturbing cyclin and Ras expression

Zoledronic acid (ZOL) was shown earlier to prolong survival in animal models of lung cancer. The aim of this study was to examine whether alteration of intracellular cyclins, cyclin-dependent kinases, cyclin-dependent kinase inhibitors, retinoblastoma, and Ras protein expression and E2F localization are among the possible antilung cancer mechanisms driven by ZOL. Furthermore, we used geranylgeraniol to test whether the mevalonate pathway is involved in the antitumor effects of ZOL against lung cancer. Line-1 cells, a murine lung adenocarcinoma cell line, were examined. ZOL significantly slowed the growth of these cells both in vitro and in vivo. The ZOL-treated cells typically arrested at the S/G2/M phase of the cell cycle, accompanied by increased intracellular levels of cyclin A, B1, and CDC2 and decreased levels of cyclin D, p21, p27, phosphorylated retinoblastoma, and Ras. In addition, ZOL affected the distribution of E2F. When geranylgeraniol was added to the ZOL-treated cells, either in vitro or in vivo, tumor growth, cell-cycle progression, the expression of certain cyclins, and cyclin-related regulatory proteins were partially returned to that of untreated controls. Therefore, ZOL elicits cell-cycle prolongation that seems to be associated with alterations in the levels of certain cyclins and cyclin-related regulatory proteins. Furthermore, the mevalonate pathway regulates ZOL-induced murine lung cancer inhibition both in vitro and in vivo.