BACKGROUND: Immunotherapy is another treatment modality for various cancers. There is little information on the antitumor effects of immunotherapy on implanted lung cancer mouse models. Toxoplasma gondii is able to potently induce a nonspecific stimulation of the host immune system. Therefore, this study evaluated the antitumor and antimetastatic effect of nonspecific immune stimulation by T. gondii in a Lewis lung cancer mouse model. METHODS: Femals C57BL/6 mice were injected with either Lewis lung cancer cells (1 X 10(6) per mouse) or 5 cysts from the T. gondii Me49 strain with various schedules. The number of survival days, the tumor size of the implanted muscle and the histopathological findings of each group were noted. In addition to these mice, the Toxoplasma antigen(50 micro gram per mouse) or a lymphokine (0.5ml per mouse) was added to boost the immunotherapy. RESULTS: No mouse in the Toxoplasma-infected group had died, whereas the mice receiving only the cancer cells (cancer control) survived for 29.1+/-4.4days. Cancer cells were revealed from 1 week after cancer cell inoculation in the muscle and from 3 weeks in the lung of the cancer control, whereas cancer cells were found in both the preinfection control and coinfection control groups from 2 weeks and 4 weeks in the lung, respectively. The in the number of survival days were 32.4+/-3.3 in the mice receiving T. gondii 2 weeks prior to the cancer cells inoculation (preinfection control), 30.9+/-5.1 in mice received both simultaneously (coinfection control), and 34.9+/-2.9 in mice received T.gondii 2 weeks after cancer cells implantation (postinfection control). These 3 infection groups had significantly longer survival days and suppressed tumor growth than those of the cancer control. In addition to these mice, and injection with the Toxoplasma antigen alone or in combination with lymphokine resulted in a significant increase in the number of survival days. CONCLUSIONS: These findings suggest that an injection with T.gondii can induce the antitumor and antimetastatic effects in Lewis lung cancer mouse models. Moreover, these effects were increased with an injection of the Toxoplasma antigen alone or in combination with lymphokine. However, this therapy can not prevent the development of cancer.
Animals ; Appointments and Schedules ; Carcinoma, Lewis Lung* ; Coinfection ; Immune System ; Immunotherapy ; Lung ; Lung Neoplasms ; Mice* ; Toxoplasma*

The incidence of lung cancer has rapidly increased and cancer patients at a later cancer stage frequently suffer from unbearable cancer-associated pain. However, the pathophysiology of lung cancer pain has not been fully described due to a lack of appropriate animal models. This study was designed to determine the effect of Lewis lung carcinoma (LLC) cell inoculation on formalin-induced pain behavior and spinal Fos expression in C57BL/6 mice. LLC cells (1.5 × 10⁵, 2.5 × 10⁵, 3.0 × 10⁵ or 5.0 × 10⁵) were inoculated into back or peri-sciatic nerve areas. Back area inoculation was adopted to determine the effect of cancer cell circulating factors and the peri-sciatic nerve area was used to evaluate the possible effects of cancer cell contacting and circulating factors on formalin-induced pain. At postinoculation day 7, LLC cell (5.0 × 10⁵) inoculations in both back and peri-sciatic nerve area significantly increased formalin-induced paw-licking time and spinal Fos expression over those in cell-media-inoculated (control) mice. Enhanced pain behavior and spinal Fos expression were significantly suppressed by ibuprofen pretreatment (250 mg/kg). The results of this study suggest that LLC cell circulating factors and inflammatory responses may be critical in enhancing pain sensation in the early stage of lung cancer cell inoculation.
Animals ; Carcinoma, Lewis Lung* ; Formaldehyde ; Humans ; Ibuprofen ; Incidence ; Lung Neoplasms ; Mice* ; Models, Animal ; Sensation

PURPOSE: Photodynamic therapy was used to lung cancer. We have made a light microscpic study on the effects of photodynamic therapy to tumor graft in skin of mice, when the power density was 600 mW/cm2 with reducing time. MATERIALS AND METHODS: These studies had been performed on sixteen C57BL/6 mice that Lewis lung carcinoma cells had been implanted. All mice were divided into four groups. One of four groups received Photogem 3 mg/kg intravenously 24 hours prior to exposure of tumor to 180 J/cm2 laser light vertically at a wavelength 635etam with a higher power density of 600 mW/cm2 than that of 400 mW/cm2 clinically. One of these group received only Photogem. The others not received Photogem and one of these irradiated with laser. The light source was the wavelength of 635 etam Diode Laser (Laxcell 2004, Bio- Optics. co. Korea) After photodynamic therapy was finished, staining and analysing of tumors were used to determine the natures and extents of injury. RESULTS: Grossly response was not observed. Histologically, there were loss of endothelium from small vessel at tumor and muscle with thrombus formation. There were focal necrosis with infiltration of inflammatory cells at tumor and adjacent tissues that irradiated with laser, regardless of administration of Photogem. CONCLUSION: Photodynamic therapy using Photogem and LASER with power density of 600 mW/cm2 destroy not only tumors incompletely but also adjacent normal tissue.
Animals ; Carcinoma, Lewis Lung* ; Endothelium ; Lasers, Semiconductor ; Lung Neoplasms ; Mice* ; Necrosis ; Photochemotherapy* ; Skin ; Thrombosis ; Transplants*

No abstract available.
Animals ; Bacteria* ; Carcinoma, Lewis Lung* ; Lactic Acid* ; Mice* ; Sarcoma 180* ; Sarcoma*

Cancer is one of the leading causes of morbidity and mortality worldwide; therefore there is a need to discover new therapeutic modules with improved efficacy and safety. Immune-(cell) therapy is a promising therapeutic strategy for the treatment of intractable cancers. The effectiveness of certain chemotherapeutics in inducing immunogenic tumor cell death thus promoting cancer eradication has been reported. Ginsenoside Rg3 is a ginseng saponin that has antitumor and immunomodulatory activity. In this study, we treated tumor cells with Rg3 to verify the significance of inducing immunogenic tumor cell death in antitumor therapy, especially in DC-based immunotherapy. Rg3 killed the both immunogenic (B16F10 melanoma cells) and non-immunogenic (LLC: Lewis Lung Carcinoma cells) tumor cells by inducing apoptosis. Surface expression of immunogenic death markers including calreticulin and heat shock proteins and the transcription of relevant genes were increased in the Rg3-dying tumor. Increased calreticulin expression was directly related to the uptake of dying tumor cells by dendritic cells (DCs): the proportion of CRT+ CD11c+ cells was increased in the Rg3-treated group. Interestingly, tumor cells dying by immunogenic cell death secreted IFN-gamma, an effector molecule for antitumor activity in T cells. Along with the Rg3-induced suppression of pro-angiogenic (TNF-alpha) and immunosuppressive cytokine (TGF-beta) secretion, IFN-gamma production from the Rg3-treated tumor cells may also indicate Rg3 as an effective anticancer immunotherapeutic strategy. The data clearly suggests that Rg3-induced immunogenic tumor cell death due its cytotoxic effect and its ability to induce DC function. This indicates that Rg3 may be an effective immunotherapeutic strategy.
Animals ; Apoptosis ; Calreticulin ; Carcinoma, Lewis Lung ; Cell Death* ; Dendritic Cells ; Heat-Shock Proteins ; Immunotherapy* ; Melanoma ; Mortality ; Panax ; Saponins ; T-Lymphocytes

BACKGROUND: Interleukin-12 (IL-12) can induce antitumor effects in vivo. This antitumor effect is associated with T cell infiltration but the effect of IL-12 on the steps of T cell migration into the tumor tissue has not been fully elucidated. This study focused on the effect of IL-12 on the tumor growth and the metastasis and on the expression of E-selectin, an adhesion molecule which is activated endothelial specific in its expression. In addition, we studied whether the expression of E-selectin is associated with the TNF-alpha, a cytokine that its production is increased by IL-12 and has functions inducing a variety of adhesion molecules. METHODS: Mice of C57BL/6 strain were injected with Lewis lung cancer cells followed by either IL-12, TNF-alpha, or normal saline by intraperitoneal route. Twenty eight days after tumor cell inoculation, metastatic nodules of lung were enumerated and immunohistochemical staining of the subcutaneous tumors were performed with monoclonal antibodies to CD4, CD8, CD16, and E-selectin. RESULTS: In IL-12 treated mice, the subcutaneously implanted Lewis lung tumors were decreased in size and the metastases were also decreased in number compared to control mice. On tumor tissues, increased infiltration of CD4+, CD8+, and CD16+ cells were observed in IL-12 treated mice compared to control mice. In control mice, E-selectin was absent on tumor vessels, but the expression of E-selectin was increased on tumor vessels of IL-12 treated mice. Administration of TNF-alpha increased not only the expression of E-selectin but also infiltrations of CD4+, CD8+, and CD16+ cells on tumor tissues. CONCLUSIONS: These results demonstrate that IL-12 inhibits tumor growth and metastases through infiltrations of inflammatory cells in mouse model of Lewis lung carcinoma and E-selectin may play a role in inflammatory cell recruitment on tumor tissue following IL-12 administration. Also, TNF-alpha may have a role as a mediator responsible for the IL-12 induced expression of E-selectin.
Animals ; Antibodies, Monoclonal ; Carcinoma, Lewis Lung* ; Cell Movement ; E-Selectin* ; Interleukin-12* ; Lung ; Lung Neoplasms ; Mice* ; Neoplasm Metastasis ; Tumor Necrosis Factor-alpha

BACKGROUND: The antitumor effects of herpes simplex virus thymidine kinase(HSV-tk) and ganciclovir(GCV) strategies for cancer gene therapy have a the following advantages:1) a direct cytotoxicity to HSV-tk modified cancer cells by GCV 2) a cell death by the local transfer of toxic metabolites from the HSV-tk modified cells to nearby unmodified tumor cells(bystander effect), and 3) in vivo bystander effect such as antitumor-immunity. Retroviral and adenoviral sequences can silence transgene expression in cells and mice. In this study, we investigated the above described advantages of HXV-tk/GCV strategy in Lewis lung cell and mouse lung cancer model using retroviral vector and adenoviral vector. Also, we observed whether the expression of a silenced gene can be reactivated by treating cell with butyrate. METHODS: Retrovirus-HSV-tk and adenovirus-HSV-tk vectors were used for the transduction of Lewis lung carcinoma(LLC) cells. The change of HSV-tk expression by butyrate was measured by Western blot.The antitumor activities containing bystander effect were observed in vivo(by MTT assay) and in vivo tumor models of various combinations of LLC and LLC-tk. RESULTS: 1. Butyrate induced the enhancement of HSV-tk expression from adenovirally transduced cells but not from retrovirally transduced cells. 2. Both retrovirus-HSV-tk and adenovirus-HSV-tk vectors with GCV treatment were effective for killing of tumor cell in vitro and suppression of LLC tumorigenicity. Bystander effect was responsible for killing of mixture of LLC-tk and LLC in vitro and in vivo-tumorigenicity model. CONCLUSION: Butyrate could augment adenoviral vector seems to be an effective approach for lung cancer therapy.
Adenoviridae ; Animals ; Butyrates ; Bystander Effect ; Carcinoma, Lewis Lung* ; Cell Death ; Genes, Neoplasm ; Genetic Therapy* ; Herpes Simplex* ; Homicide ; Lung ; Lung Neoplasms ; Mice* ; Phosphotransferases* ; Retroviridae ; Simplexvirus* ; Thymidine ; Transgenes ; Zidovudine*

BACKGROUND AND OBJECTIVEThe mouse lung cancer orthotopic model includes spontaneous lung cancer model and endotracheal transplanted model, and etc. The spontaneous lung cancer needs longer time and does not ensure the rate of the generation of the tumor; as for endotracheal transplanted model, the position and size of the tumor are instable. In this study, the 3LL cell line was orthotopically transplanted into the lung of the C57BL/6 mice, compare to the heterotopic model, to discuss their stability and transfer-characteristics. And this study was also to optimize the method of establishing lung cancer orthotopic animal model.

METHODSDifferent quantity of 3LL cells were inoculated into the left oxter of C57BL/6 mice to establish the heterotopic model; or suspended with Matrigel then inoculated into the left lung of C57BL/6 mice to establish orthotopic model. The survival-time of the mice was examined. The tissue was collected for the subsequent histology assay after euthanizing the mice. Microvessels density (MVD) was observed and counted by immunohistological chemistry. CD44v was detected by flow cytometry.

RESULTSTTumor-form-rate of the heterotopic group were 100%, 66.7%, 16.7%, respectively, and had no macroscopic transfer. Tumor-form-rate of the orthotopic group were 100%, 100%, 83.3%, respectively, and had widespread transfer in contralateral chest and the lung. The median survival time of the orthotopic group (38, 35, 23 days) were less than the heterotopic group (82, 72, 50 days). MVD of the orthotopic group (120.2 +/- 9.73) was higher than the heterotopic group (92.6 +/- 7.12). The expression of CD44v of orthotopic (26.46 +/- 1.56)% was higher than the heterotopic group (23.13 +/- 1.02)%.

CONCLUSIONThe lung cancer orthotopic model which established by 3LL cells transplanted into the lung of the mice is simple, dependable, repeatable and has stronger transfer characteristics than the heterotopic model.

Animals ; Carcinoma, Lewis Lung ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Lung Neoplasms ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Random Allocation

Objective This study aimed to establish a pre-metastatic niche mouse model utilizing luciferase-labeled Lewis (Luc-Lewis) lung cancer cells and to assess the efficacy of this model employing both qualitative and quantitative methods. Methods C57BL/6 mice were categorized into two groups: a normal control group and a model group, each containing 15 individual mice. The pre-metastatic niche model was established via tail vein injection of Luc-Lewis lung cancer cells. Body mass were measured daily for all groups. Tumor fluorescence signals within the mice were detected using a high-throughput enzyme marker instrument. Lung tissue specimens were harvested to evaluate metastatic progression. HE staining was used to assess histopathological changes. Real-time quantitative PCR and Western blot analysis were used to detect the mRNA and protein expression of lysyl oxidase (LOX), matrix metalloproteinase 9 (MMP9), versican (VCAN), and fibronectin (FN), which are the specific markers for the formation of the microenvironment of lung tissues before metastasis. Results Significant declines in body mass and observable lethargy were noted in the model group when compared to the control group. Distinct fluorescence signals were observed in the lung tissue of the model group, demonstrating a positive correlation with the duration of model establishment. By day 14, elevated mRNA and protein expression levels of LOX, MMP9, VCAN, and FN were significantly evident. In addition, histopathological evaluations revealed augmented interstitial thickness, alveolar atrophy and significant inflammatory cell infiltration within the lung tissues of the model group. By the 21st day, metastatic lesions manifested in the lung tissues of the model group, suggesting an approximate pre-metastatic niche maturation timeline of 14 days. Conclusion A pre-metastatic niche mouse model for Lewis lung cancer is successfully established.
Mice ; Animals ; Lung Neoplasms/pathology* ; Matrix Metalloproteinase 9 ; Mice, Inbred C57BL ; Carcinoma, Lewis Lung ; Disease Models, Animal ; RNA, Messenger ; Tumor Microenvironment

OBJECTIVETo test the hypothesis that malignant tumor metastasis is mediated also through a non-cellular, essentially molecular, mechanism in addition to the cellular pathway.

METHODSThe sex-determining region on the Y chromosome was detected as the marker of the primary tumors using PCR in Lewis lung carcinoma (LLC) in vitro and in female C57BL/6 mice bearing LLC with spontaneous metastasis. The macroscopic and microscopic metastases in the tumor-bearing mice were examined for SRY expression by PCR and in situ hybridization, using the tissues from male and female mice as the positive or negative controls.

RESULTS AND CONCLUSIONPositive SRY gene expression was detected in the metastatic foci in the LLC-bearing female mice, suggesting the origination of these tumor cells from the primary tumor foci. We have failed to verify the non-cellular metastasis hypothesis in this animal experiment, but given the limitations of this experiment, we consider further investigation still necessary for verification of this hypothesis using other methods.

Animals ; Carcinoma, Lewis Lung ; genetics ; pathology ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Neoplasm Transplantation ; Sex-Determining Region Y Protein ; genetics ; metabolism