The role of prostaglandin E2 (PGE2) in directly stimulating metastatic spread by Lewis lung carcinoma (LLC) cells was examined with the use of an in vitro migration model for tumor dissemination. The extent to which cloned metastatic and nonmetastatic LLC cells migrated out of glass capillary tubes in vitro reflected their capacity to form pulmonary metastases in vivo. The addition of PGE2 to metastatic LLC cells further stimulated their migration. Other cyclooxygenase products, besides PGE2, did not stimulate the migration of metastatic LLC cells. Nonmetastatic LLC cells did not migrate out of capillary tubes, even in the presence of exogenous PGE2. the amount of PGE2 secreted by cloned LLC cells was quantitated by a radioimmunoassay. Nonmetastatic LLC cells secreted more PGE2 than did the metastatic LLC cells. When the nonmetastatic LLC cells were either mixed with or placed adjacent to cloned metastatic LLC cells, the migration by the metastatic LLC cells was stimulated. The ...

Rigorous and systematic pre-clinical studies are necessary and essential to establish the efficacy and safety of Oriental herbs and formulas in order to transform traditional herbal practices into evidence-based medicine. Here we evaluated the anti-cancer activities of the ethanol extract of Ka-mi-kae-kyuk-tang (KMKKT), a formula of ten Oriental herbs, with a battery of in vitro and in vivo mechanism-based biomarkers involving angiogenesis, apoptosis and metastasis. The results show that KMKKT suppressed the vascular endothelial responses by inhibiting basic fibroblast growth factor (bFGF)-induced ERK1/2 phosphorylation, cell migration as well as tube formation in the human umbilical vein endothelial cell model, and decreased the hypoxia-induced HIF1α and vascular epithelial growth factor (VEGF) expression in the mouse Lewis lung carcinoma (LLC) cells in vitro, and inhibited the bFGF-induced angiogenesis in chick chorioallantoic membrane model, and in the Matrigel plugs in mice. Intraperitoneal ...

Evidence-Based Complementary and Alternative Medicine (eCAM) is an international peer-reviewed, Open Access journal that seeks to understand the sources and to encourage rigorous research in this new, yet ancient world of complementary and alternative medicine.

Abstract: Several classes of flavonoids [flavanoids (1-10), flavonol (11), isoflavones (12-18), isoflavanones (19-22), isoflavans (23-26), chalcones (27-30), auronol (31), pterocarpans (32-37), 2-arylbenzofuran (38), and neoflavonoid (39)] and lignans (40-42) isolated from the MeOH extract of Brazilian red propolis were investigated for their cytotoxic activity against a panel of six different cancer cell lines including murine colon 26-L5 carcinoma, murine B16-BL6 melanoma, murine Lewis lung carcinoma, human lung A549 adenocarcinoma, human cervix HeLa adenocarcinoma, and human HT-1080 fibrosarcoma cell lines ...

The surface exposure of CRT and ERp57 was increased in CT26 clones derived from cells transiently exposed to nocodazole that contained close to twice the DNA content of parental cells (which we refer to as hyperploid cells), although the surface expression of most other membrane proteins was unaltered (Fig. 1D and fig. S8). This hyperploidy-associated increase in CRT exposure was also observed in mouse Lewis lung carcinoma (LLC) and fibrosarcoma MCA205 cells, as well as in human cancer cell lines (fig. S9). As compared to their parental counterparts, hyperploid clones exhibited constitutive PERK and eIF2α phosphorylation (Fig. 1E). Interruption of the CRT exposure pathway reduced the clonogenic potential of hyperploid cells (Fig. 1, F and G), suggesting a functional link between the ER stress-associated CRT exposure pathway and the fitness of hyperploid cells.. Because hyperploidization is linked to CRT exposure, we wondered whether cancer cells with increased DNA content might be subjected ...

The surface exposure of CRT and ERp57 was increased in CT26 clones derived from cells transiently exposed to nocodazole that contained close to twice the DNA content of parental cells (which we refer to as hyperploid cells), although the surface expression of most other membrane proteins was unaltered (Fig. 1D and fig. S8). This hyperploidy-associated increase in CRT exposure was also observed in mouse Lewis lung carcinoma (LLC) and fibrosarcoma MCA205 cells, as well as in human cancer cell lines (fig. S9). As compared to their parental counterparts, hyperploid clones exhibited constitutive PERK and eIF2α phosphorylation (Fig. 1E). Interruption of the CRT exposure pathway reduced the clonogenic potential of hyperploid cells (Fig. 1, F and G), suggesting a functional link between the ER stress-associated CRT exposure pathway and the fitness of hyperploid cells.. Because hyperploidization is linked to CRT exposure, we wondered whether cancer cells with increased DNA content might be subjected ...

Authors: Riordan NH, Meng X, Riordan HD.. ABSTRACT: Recruitment of new blood vessels plays a crucial role in tumor survival and growth. Several agents that act as angiogenesis inhibitors are currently being investigated as anti-tumor agents. Proteoglycan extract (PGM) was tested for anti-angiogenic, immunostimulatory, and anti-neoplastic activity. PGM is a non-toxic extract of the ubiquitous plant, Convolvulus arvensis. In the chicken egg chorioallantoic membrane assay PGM inhibited new blood vessel growth in a dose-dependent manner. Results were 18, 55, and 73% inhibition at concentrations of 50, 100, and 200 mcg, respectively. PGM significantly inhibited tumor growth in the mouse fibrosarcoma (S-180 Kun Ming 3-4 wk old mixed male/female, 10 animals per group, 250-1000 mcg daily doses for 14 days), and mouse Lewis lung carcinoma (C57, 6 wk old mixed male/female, 10 animals per group, 250-1000 mcg daily doses for 14 days) models. Inhibition (54-77% inhibition by weight compared to controls, up ...

R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv, Ukraine. [email protected]. Introduction: It is known that the tumor growth is accompanied by changes of redox carriers in the mitochondrial electron transport chain. These changes may result in decrease of energy metabolism. The functional state of the mitochondrial electron transport chain also defines the intensity of production and accumulation of the oxygen and nitrogen reactive species (ROS and NOS), which affect different biological processes and can promote cell damage. It has already been observed that non-cytotoxic doses of oxyresveratrol (OXY) showed significant antineoplastic and anti-metastatic effects and prolonged the life of animals with tumors. Aim: To analyze the influence of OXY on the mitochondrial iron-sulfur centers and nitrosyl-hem complexes of tumor cells during Lewis lung carcinoma growth. Materials and Methods: EPR method was used to analyze the OXY influence on the ...

Background: Lung cancer is the most common human cancer. There are numerous studies that have sought to discover genetic impact factors associated with it. The study reported on in this article is a systematic search for the genetic networks underlying the Lewis lung carcinoma (LLC) model.Methods: LLC-Gene relation data were extracted from the ResNet 11 Mammalian database, containing 175 LLC candidate genes (nodes). Pathway Enrichment Analysis, Sub-Network Enrichment Analysis, Network Connectivity Analysis, and Network Metrics Analysis were conducted to study network attributes and select the top nodes (genes). LLC-Drug and Drug-Gene relation data examined LLC-Gene relations at the small molecule level.Results: A total of 166 of 175 genes that were enriched in 134 LLC candidate pathways (p < 1e-07), demonstrated strong gene-gene interactions. Metrics analysis revealed 4 genes, IL6, TNF, VEGFA, and HIF1A, as top LLC candidates as measured by replication frequency, network centrality, and functional

Id1 promotes carcinogenesis and metastasis, and predicts prognosis of non-small cell lung cancer (NSCLC)-adenocarcionoma patients. We hypothesized that Id1 may play a critical role in lung cancer colonization of the liver by affecting both tumor cells and the microenvironment. Depleted levels of Id1 in LLC (Lewis lung carcinoma cells, LLC shId1) significantly reduced cell proliferation and migration in vitro. Genetic loss of Id1 in the host tissue (Id1(-/-) mice) impaired liver colonization and increased survival of Id1(-/-) animals. Histologically, the presence of Id1 in tumor cells of liver metastasis was responsible for liver colonization. Microarray analysis comparing liver tumor nodules from Id1(+/+) mice and Id1(-/-) mice injected with LLC control cells revealed that Id1 loss reduces the levels of EMT-related proteins, such as vimentin. In tissue microarrays containing 532 NSCLC patients samples, we found that Id1 significantly correlated with vimentin and other EMT-related proteins. Id1 ...

Inflammatory angiogenesis is a critical process in tumor progression and other diseases. The inflammatory cytokine IL-1β promotes angiogenesis, tumor growth, and metastasis, but its mechanisms remain unclear. We examined the association between IL-1β-induced angiogenesis and cell inflammation. IL-1β induced neovascularization in the mouse cornea at rates comparable to those of VEGF. Neutrophil infiltration occurred on day 2. Macrophage infiltration occurred on days 4 and 6. The anti-Gr-1 Ab-induced depletion of infiltrating neutrophils did not affect IL-1β- or VEGF-induced angiogenesis. The former was reduced in monocyte chemoattractant protein-1-deficient (MCP-1-/-) mice compared with wild-type mice. After day 4, clodronate liposomes, which kill macrophages, reduced IL-1β-induced angiogenesis and partially inhibited VEGF-induced angiogenesis. Infiltrating macrophages near the IL-1β-induced neovasculature were COX-2 positive. Lewis lung carcinoma cells expressing IL-1β (LLC/IL-1β) ...

Cancer-associated inflammation contributes to tumor progression and metastasis. Defective dendritic cell (DC) function causes impaired adaptive immunity to antigens expressed by tumors. We recently found that the intra-tumor inflammatory milieu recruits and skews development of DC precursors (pre-cDC) towards a novel Gr-1+ subpopulation, which has tolerogenic properties. This transformation reduced the proliferation and expansion of antigen-specific cytotoxic T cells in tumors. We sought to identify cytokines that regulate this differentiation. Isolated pre-cDC from mice spleen was incubated with tumor-conditioned medium (TCM) derived from Lewis lung carcinoma cell line. Supernatants from cultures were collected to analyze cytokine levels. Expression of Gr-1+ on DC was monitored by flow cytometry. We found that TCM induced pre-cDC to differentiate into Gr-1+ cDC. Neutralizing anti-cytokine Ab studies revealed that IL-6 play a role in promoting Gr-1+ cDC differentiation. Pre-cDC from IL-6-/- mice ...

The lymphatic system is an important route for cancer dissemination, and lymph node metastasis (LNM) serves as a critical prognostic determinant in cancer patients. We investigated the contribution of COX-2-derived prostaglandin E2 (PGE2) in the formation of a premetastatic niche and LNM. A murine model of Lewis lung carcinoma (LLC) cell metastasis revealed that COX-2 is expressed in DCs from the early stage in the lymph node subcapsular regions, and COX-2 inhibition markedly suppressed mediastinal LNM. Stromal cell-derived factor-1 (SDF-1) was elevated in DCs before LLC cell infiltration to the lymph nodes, and a COX-2 inhibitor, an SDF-1 antagonist, and a CXCR4 neutralizing antibody all reduced LNM. Moreover, LNM was reduced in mice lacking the PGE2 receptor EP3, and stimulation of cultured DCs with an EP3 agonist increased SDF-1 production. Compared with WT CD11c+ DCs, injection of EP3-deficient CD11c+ DCs dramatically reduced accumulation of SDF-1+CD11c+ DCs in regional LNs and LNM in ...

Characterization of the stem-like properties of cancer stem cells (CSCs) remain indirect and qualitative, especially the ability of CSCs to undergo asymmetric cell division for self renewal and differentiation, a unique property of cells of stem origin. It is partly due to the lack of stable cellular models of CSCs. In this study, we developed a new approach for CSC isolation and purification to derive a CSC-enriched cell line (LLC-SE). By conducting five consecutive rounds of single cell cloning using the LLC-SE cell line, we obtained two distinct sub-population of cells within the Lewis lung cancer CSCs that employed largely symmetric division for self-renewal (LLC-SD) or underwent asymmetric division for differentiation (LLC-ASD ...

Figure 4. PFK15 suppresses the growth, metastatic spread, and glucose metabolism of LLC tumors in syngeneic mice. Groups of 8 C57Bl/6 mice (20 g) were injected s.c. with 0.1 mL of an LLC cell suspension (1 × 106 cells) and tumor masses were determined in a blinded fashion with Vernier calipers. Mice bearing xenografts (150-200 mg) then were randomized to DMSO or PFK15 and microcaliper measurements (A) and body mass (B) were collected daily. After 14 days, mice were euthanized and the lungs were examined for metastases (C). Tumors were extracted in a separate subgroup of mice after 4 days and analyzed for F26BP (D) and cleaved caspase 3 expression as a measure of apoptosis (E and F). A separate cohort of tumor-bearing mice underwent baseline micro-PET imaging of FDG uptake and, 24 hours later, were administered PFK15 (25/kg i.p. once) and the micro-PET scan was repeated in 45 minutes. Regions of interest in the tumor and cerebellum were quantified in quadruplicate (G) and a representative cut is ...

We next tested the prediction that the inhibitory activity of LLC CD4+CD25+ TILs should be greater than that of peripheral LN CD4+CD25+ cells. As shown in Fig. 3⇑B, there was no difference in the suppressive potency of Tregs over various ratios of T effector (Teff) to Treg cells from peripheral LNs of tumor-bearing and control mice. In contrast, TIL CD4+CD25+ cells retrieved from LLC tumors exhibited far more potent suppressive activity than LN CD4+CD25+ cells from tumor-free mice (Fig. 3⇑C). Thus, the suppressive effects of peripheral LN Tregs and TIL Tregs correlated well with their levels of TNFR2 expression. As shown in Fig. 3⇑, B and C, the potent inhibitory effect exerted by CD4+CD25+ TILs was not Ag specific because both targeted responder cells and APCs were from tumor-free mice. The phenotype of TIL CD4+CD25+ cells, which were 75∼100% TNFR2+, resembled that of normal mouse LN TNFR2+ Tregs and were indicative of an activated/memory subset (data not shown).. Although other factors ...

A carcinoma discovered by Dr. Margaret R. Lewis of the Wistar Institute in 1951. This tumor originated spontaneously as a carcinoma of the lung of a C57BL mouse. The tumor does not appear to be grossly hemorrhagic and the majority of the tumor tissue is a semifirm homogeneous mass. (From Cancer Chemother Rep 2 1972 Nov;(3)1:325) It is also called 3LL and LLC and is used as a transplantable malignancy ...

The first key takeaway for how to start an LLC quickly is that you should avoid any LLC structure that does not provide for succession. Any self-employed individual who wishes to establish an LLC has to follow the same process as any business owner who wants to pass control of his or her firm to another individual. In the case of an LLC, the person passing control (the founder or director) retains the power and authority to run the business. He or she can select the members he wants to retain, hire employees, get involved in all business decisions, and control the day-to-day operation. Therefore, an LLC provides no personal asset protection for the founder of directors. ...

An investigation of the ill vitro toxicities of a group of commercial chemicals and formulations revealed very poor ill vivo/in vitro correlations. Some were toxic to the 3T3-Ll cells, yet of very low toxicity to rats. This was partly due to the poor solubility of some of the substances, which probably caused their virtual non-toxicity to rats by oral dosage. Chemical volatility is another methodological problem for ill vitro assays. A simple modification of the FRAME KB cytotoxicity assay was successfully developed in order to prevent the underestimation of the cytotoxicities of volatile liquids ...

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