Induction of Tumor Necrosis Factor and Interleukin-6 mRNA in Human Cytotrophoblast Cells Exposed to Lipopolysaccharide

Objective: The cytokines tumor necrosis factor (TNF) and interleukin-6 (IL-6) have previously been identified in placental tissue and are known to be mediators of infection-associated induction of the host immune system. This study was undertaken to better characterize the in vitro regulation of these cytokines in cytotrophoblast cells when challenged with the bacterial product lipopolysaccharide (LPS)

H19-Promoter-Targeted Therapy Combined with Gemcitabine in the Treatment of Pancreatic Cancer

Pancreatic cancer is the eighth cancer leading cause of cancer-related death in the world and has a 5-year survival rate of 1–4% only. Gemcitabine is a first line agent for advanced pancreatic therapy; however, its efficacy is limited by its poor intracellular metabolism and chemoresistance. Studies have been conducted in an effort to improve gemcitabine treatment results by adding other chemotherapeutic agents, but none of them showed any significant advantage over gemcitabine monotherapy. We found that 85% of human pancreatic tumors analyzed by in situ hybridization analyses showed moderated to strong expression of the H19 gene. We designed a preclinical study combining gemcitabine treatment and a DNA-based therapy for pancreatic cancer using a non viral vector BC-819 (also known as DTA-H19), expressing the diphtheria toxin A chain under the control of the H19 gene regulatory sequences. The experiments conducted either in an orthotopic and heterotopic pancreatic carcinoma animal model showed better antitumor activity following the sequential administration of the vector BC-819 and gemcitabine as compared to the effect of each of them alone. The results presented in the current study indicate that treatment with BC-819 in combination with gemcitabine might be a viable new therapeutic option for patients with advanced pancreatic cancer

Aerosolized BC-819 Inhibits Primary but Not Secondary Lung Cancer Growth

Despite numerous efforts, drug based treatments for patients suffering from lung cancer remains poor. As a promising alternative, we investigated the therapeutic potential of BC-819 for the treatment of lung cancer in mouse tumor models. BC-819 is a novel plasmid DNA which encodes for the A-fragment of Diphtheria toxin and has previously been shown to successfully inhibit tumor growth in human clinical study of bladder carcinoma. In a first set of experiments, we examined in vitro efficacy of BC-819 in human lung cancer cell-lines NCI-H460, NCI-H358 and A549, which revealed >90% reduction of cell growth. In vivo efficacy was examined in an orthotopic mouse xenograft lung cancer model and in a lung metastasis model using luminescent A549-C8-luc adenocarcinoma cells. These cells resulted in peri- and intra-bronchiolar tumors upon intrabronchial application and parenchymal tumors upon intravenous injection, respectively. Mice suffering from these lung tumors were treated with BC-819, complexed to branched polyethylenimine (PEI) and aerosolized to the mice once per week for a period of 10 weeks. Using this regimen, growth of intrabronchially induced lung tumors was significantly inhibited (p = 0.01), whereas no effect could be observed in mice suffering from lung metastasis. In summary, we suggest that aerosolized PEI/BC-819 is capable of reducing growth only in tumors arising from the luminal part of the airways and are therefore directly accessible for inhaled BC-819

Expression of the H19 Oncofetal Gene in Premalignant Lesions of Cervical Cancer: A Potential Targeting Approach for Development of Nonsurgical Treatment of High-Risk Lesions

Background. Recent data suggest a role for H19 gene in promoting cancer transformation and progression. Cervical cancer, progresses from high-grade lesions (CIN3). At present, it is unclear if CIN lesions express H19. Objectives. To determine H19 expression in patient samples of CIN3 as well as the ability of a construct in which the promoter from the H19 gene drives expression of the diphtheria toxin A chain (DTA) to inhibit cervical cancer cell growth in vitro. Methods. H19 transcript levels were evaluated on 10 biopsies of CIN3 using in situ hybridization. PCR was used to examine H19 expression in cervical cancer cell lines and in two samples from a patient with cervical carcinoma. Cell lines were transfected with H19-DTA to determine its impact on cell number. Results. H19 gene was expressed in the area of CIN3 in 9 out of 10 samples. RT-PCR indicated expression of H19 in cervical cancer samples and in one of the three cell lines examined. Transfection of all cell lines with H19-DTA vector resulted in inhibited cell growth. Conclusions. H19 is expressed in the majority of CIN3 samples. These results suggest that most CIN3 lesions could be targeted by H19-DTA. Further in vivo preclinical studies are thus warranted.Peer Reviewe

Histology of orthotopically induced lung tumors.

<p>Tumors were detectable as early as 10 days after intrabronchial implantation in the peribronchiolar region of the lungs (A). End stages of intrabronchially induced lung tumors were characterized by multiple, peribronchiolar and parenchymal localized tumors (B) and by tumor tissue within the luminal lung regions as well (C). Intravenous injection of A549-C8-luc, however, resulted in tumors predominately growing in the parenchyma of the lungs but not peribronchiolar or within the bronchi (D). Figure D shows parenchymal tumor tissue close to a bronchus, but not peribronchiolar. Alveolar tissue localized between the tumor and the bronchus however is condensed by the adjacent tumor nodule.</p

Survival of untreated and treated tumor-bearing mice.

<p>Mice bearing intrabronchially induced lung tumors showed a significant survival benefit (p = 0.01) when treated with BC-819 compared to animals which received no treatment (A). No survival benefit was observed in mice suffering from intravenously induced lung tumors, independent of treatment with BC-819 (B).</p

Effect of BC-819 on cell growth in different lung cancer cell lines.

<p>NCI-H460, NCI-H358 and A549 cells were transfected, using Lipofectamine 2000, with BC-819 and co-transfected with a plasmid, encoding for the reporter enzyme luciferase. As early as 24 h after transfection, luciferase activity (indirectly indicating cell growth) was reduced by at least >90% when 350 ng BC-819 was used (A). 48 hours later luciferase activity was decreased by more than 98%, except for the cell line NCI-H358. However, a luciferase decrease of more than 98% was observed in all cell lines when the amount of BC-819 was increased to 700 ng (B). The influence of BC-819 on bioluminescent A549-C8-luc was examined as well and revealed decreased luciferase and therefore reduced cell growth (more than 50%) as early as after 24 hours (C). Maximum inhibition of cell growth was reached by 48 hours after transfection, becoming apparent through more than 75% reduction in luciferase activity.</p