Mouse Squamous Cell Carcinoma Research Articles

Application of surface-enhanced Raman in skin cancer by plasma

We have developed a mouse squamous cell carcinomas (SCC) model by diniethylbenzanthracene (DMBA) and ultraviolet (UVB). A silver colloid as SERS-active substrates is used for detecting the blood plasma of mouse. The relative intensity of the band at 942 and 1499 cm−1 is higher in SCC model than in healthy one. Therefore, it can be used as an important “fingerprint” in order to diagnose these diseases. Results show us how to get high signal-to-noise ratio of biological macromolecules surface-enhanced Raman scattering spectra in blood plasma. And also offer useful help for understanding the rich molecular structure information in biological tissues. It provides a molecular spectroscopy way for early detection of disease in blood plasma.

Increased basic fibroblast growth factor release and proliferation in xenotransplanted squamous cell carcinoma after combined irradiation/anti-vascular endothelial growth factor treatment

Novel strategies of cancer therapy combine irradiation and anti-angiogenic active compounds. However, little is known concerning the undesired cellular and molecular effects caused by this novel treatment concept. We used a mouse squamous cell carcinoma (SCC) xenotransplantation model to evaluate the potential undesired effects which compromise the success of this therapeutic combination. SCCs were subcutanously implanted in nude mice. Animals were treated with a fractionated irradiation scheme (5x4Gy) alone or in combination with daily injections of anti-vascular endothelial growth factor (VEGF) antibodies. Controls remained untreated. Before and after treatment, resonance imaging (MRI), ultrasound and near-infrared spectrometry were used to evaluate tumor vessel integrity. Finally, tumors were explanted and VEGF, basic fibroblast growth factor (bFGF), vessel density, proliferation and apoptotic activity were analyzed by immunohistochemistry. Irradiation caused VEGF release and we found evidence for VEGF-mediated vessel protection. In the tumors derived from the combined treatment, blood volume was decreased, and apoptotic indices were increased. Remarkably, bFGF levels and proliferative indices were also increased. Combined irradiation/anti-VEGF treatment resulted in the desired VEGF depletion and increased tumor cell apoptosis. Nonetheless, bFGF and proliferation also increased, possibly suggesting a compensatory response. The application of additional targeted drugs may help develop more effective SCC treatments.

Multiple Bowen's Disease: In a Veteran Who Participated in Vietnam War

Dear Editor: Many etiological factors have been suggested for Bowen's Disease (BD). However, the relationship between tetrachlorodibenzo-p-dioxin (TCDD) and BD has not yet been clarified. We report here on a case detailing TCDD as a possible cause of multiple BD. A 63-year-old-man presented with multiple erythematous and scaly plaques over the whole body, which had first appeared over 10 years prior (Fig. 1). He had a history of contact with defoliants while he participated in the Vietnam War (2 years), 40 years ago and was treated for peripheral neuropathies that were possibly related to Agent Orange. The patient was otherwise in good health. The physical examination revealed six scaly erythematous papules and plaques on his back, arm, leg, and finger. According to the patient the lesions occurred spontaneously. Skin biopsies from the back and finger were performed. Histologic examination showed full thickness epidermal atypia with adnexal involvement (Fig. 2). The clinical appearance of the skin lesion and the microscopic descriptions were consistent with a diagnosis of multiple BD. Fig. 1 Multiple scaly (A) erythematous papules and plaques on the back, (B) right 4th finger, (C) left arm, and (D) right leg. Fig. 2 The skin biopsy specimen of the lower back showed full-thickness involvement by atypical keratinocytes. Large, round and hyperchromatic nuclei with mitoses were contained within the dyskeratotic cells (H&E, ×200). TCDD is a kind of dioxin isomer, and TCDD is contained in Agent Orange, which was used as a defoliant during the Vietnam War from 1961 to 1970. TCDD has a half-life of approximately 8 years in humans and it accumulates primarily in fatty tissues over time, so even small exposures may eventually reach dangerous levels1. Both the National Toxicology Programs and the International Agency for Research on Cancer have classified TCDD as a human carcinogen. Over the past several years a number of studies have described the cutaneous effects of TCDD toxicity. The common TCDD-induced skin lesions are chloracne, porphyria cutanea tarda, hyperpigmentation, and malignant fibrous histiocytomas of the skin. In Korea, 2 cases of BCC, 1 case of SCC, and 1 case of merkel cell carcinoma in Vietnam Veterans exposed to TCDD have been reported2,3. Ikuta et al.4 have reported that in primary cultured keratinocytes, TCDD increases post-confluent proliferation and late differentiation. TCDD also inhibits culture-induced senescence, which is a form of permanent cell cycle arrest. Wyde et al.5 identified the development of cutaneous papillomas and squamous cell carcinomas in mice exposed to dermal (≥ 52 ng/kg) and oral (893 ng/kg) TCDD. Thus, TCDD can modulate the differentiation of keratinocytes and promote tumor formation in the skin. BD is SCC in situ, and this has the potential to progress to invasive SCC. The known etiological factors of BD include: (1) irradiation such as sun exposure, photochemotherapy and, radiotherapy (2) carcinogens such as arsenics (3) immunosuppression such as therapeutic immune suppressions and, acquired immune deficiency syndrome, and (4) oncogenic human papilloma virus (HPV) types, such as HPV 16. Although BD has a predilection for sun-exposed areas, the BD seen in non-sun exposed areas suggest a strong association of BDs with arsenic or HPV. In the present case, PCR of the skin tissue was negative for HPV. Since our patient denied a history of arsenic exposure, we believe that the multiple BD in our case was likely associated with the TCDD exposure.

Neonatal mice do not have increased sensitivity to induction of squamous cell carcinomas

Squamous cell carcinoma (SCC) is linked with the lifelong cumulative effect of ultraviolet radiation (UVR). In contrast, epidemiological data have shown that sunburn in childhood is a stronger risk factor for cutaneous malignant melanoma than continuous UVR, indicating a higher carcinogenic sensitivity early in life. We investigated how a high neonatal dose of UVR affects the development of SCC in mice irradiated later in life. We used simulated solar radiation (sun) and solarium radiation (solarium). Ninety-nine C3.Cg/TifBomTac-immunocompetent hairless mice received 0, 25 or 35 standard erythema doses (SED) UVR when they were 4 days old followed by 4 SED sun or 4 SED solarium three times/weekly from 9 weeks of age. Tumours developed faster in mice treated with 35 SED UVR + 4 SED sun compared with 4 SED sun, but no change was observed in the cumulative dose required to achieve tumours. Tumours also developed faster in mice treated with 35 SED UVR + 4 SED solarium compared with 4 SED solarium, and a difference was also observed in the cumulative dose required to achieve tumours. If the Skin Cancer Utrecht-Philadelphia-murine spectrum was used to weigh the delivered irradiance instead of the International Commission on Illumination erythema action spectrum, tumours developed after the same accumulated dose. In conclusion, this study does not indicate increased sensitivity to induction of SCC early in life.

FAK deletion promotes p53-mediated induction of p21, DNA-damage responses and radio-resistance in advanced squamous cancer cells.

Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that is elevated in a variety of human cancers. While FAK is implicated in many cellular processes that are perturbed in cancer, including proliferation, actin and adhesion dynamics, polarisation and invasion, there is only some limited information regarding the role of FAK in radiation survival. We have evaluated whether FAK is a general radio-sensitising target, as has been suggested by previous reports. We used a clean genetic system in which FAK was deleted from mouse squamous cell carcinoma (SCC) cells (FAK −/−), and reconstituted with exogenous FAK wild type (wt). Surprisingly, the absence of FAK was associated with increased radio-resistance in advanced SCC cells. FAK re-expression inhibited p53-mediated transcriptional up-regulation of p21, and a sub-set of other p53 target genes involved in DNA repair, after treatment with ionizing radiation. Moreover, p21 depletion promoted radio-sensitisation, implying that FAK-mediated inhibition of p21 induction is responsible for the relative radio-sensitivity of FAK-proficient SCC cells. Our work adds to a growing body of evidence that there is a close functional relationship between integrin/FAK signalling and the p53/p21 pathway, but demonstrates that FAK's role in survival after stress is context-dependent, at least in cancer cells. We suggest that there should be caution when considering inhibiting FAK in combination with radiation, as this may not always be clinically advantageous.

Targeting of the Tumor Suppressor GRHL3 by a miR-21-Dependent Proto-Oncogenic Network Results in PTEN Loss and Tumorigenesis

Despite its prevalence, the molecular basis of squamous cell carcinoma (SCC) remains poorly understood. Here, we identify the developmental transcription factor Grhl3 as a potent tumor suppressor of SCC in mice, and demonstrate that targeting of Grhl3 by a miR-21-dependent proto-oncogenic network underpins SCC in humans. Deletion of Grhl3 in adult epidermis evokes loss of expression of PTEN, a direct GRHL3 target, resulting in aggressive SCC induced by activation of PI3K/AKT/mTOR signaling. Restoration of Pten expression completely abrogates SCC formation. Reduced levels of GRHL3 and PTEN are evident in human skin, and head and neck SCC, associated with increased expression of miR-21, which targets both tumor suppressors. Our data define the GRHL3-PTEN axis as a critical tumor suppressor pathway in SCC.

Cyclooxygenase-dependent signaling is causally linked to non-melanoma skin carcinogenesis: pharmacological, genetic, and clinical evidence

Cyclooxygenase (COX)-derived prostaglandins (PGs) exhibit manifold functions in acute and chronic skin inflammation induced by a number of physical (ultraviolet (UV) light, wounding) and chemical (12-O-tetradecanoylphorbol 13-acetate (TPA), arachidonic acid) noxious stimuli. Depending on the challenge and the context, constitutively expressed COX-1 or the transiently induced COX-2 isoform are of relevance. Moreover, squamous cell carcinoma (SCC) of skin is a prominent example of epithelial neoplasia that consistently overexpresses COX-2 in the parenchyme and the mesenchyme of premalignant and malignant lesions, while COX-1 expression remains unaltered. Pharmacological, clinical, and experimental animal studies as well as a few epidemiological studies document the importance of PG signaling in non-melanoma skin cancer including SCC and basal cell carcinoma (BCC) in humans and mice. Increased levels of PGE(2) and PGF(2α) in premalignant and/or malignant epithelial skin cancers are due to the constitutive upregulation of enzymes involved in PG biosynthesis, such as COX-2, and downregulation of the tumor suppressor gene 15-hydroxy-prostaglandin dehydrogenase (15-PGDH), which is involved in the inactivation of PG, thus counteracting the activities of COX. Most remarkably, genetic studies show that mice which are deficient in COX-2 or COX-1 are protected from the development of SCC when applying the multi-stage chemical carcinogenesis protocol. Conversely, the forced overexpression of COX-2 in the proliferative basal compartment of the stratified skin epidermis results in spontaneous hyperplasia and dysplasia in transgenic mice and furthermore a sensitization for cancer development by conferring an auto-promoted skin phenotype. In multi-stage carcinogenesis, it also becomes clear that aberrant COX-2 overexpression and activity are causally involved in tumor promotion and tumor progression rather than initiation. In contrast, using as inducer of carcinogenesis the complete carcinogen UV B light, depletion of COX-2 but not of COX-1 makes mouse skin resistant for SCC, indicating that here, only COX-2 is essential. Depending on the type of challenge, COX-2-dependent signaling contributes to the pre-invasive growth of the skin epidermis by a delayed onset of terminal differentiation, or stimulation of hyperproliferation and survival. With respect to BCC, the genetic ablation of COX-2 but also of COX-1 leads to a strongly reduced tumor burden in the skin of Patched (Ptch)1(+/-) mice, which due to the deletion of a Ptch1 allele, spontaneously develop BCC resembling human familial basal cell nevus syndrome and sporadic BCC. Nonsteroidal anti-inflammatory drugs and the COX-2-selective inhibitors (COXibs) exhibit impressive efficacy inhibiting tumor burden in various mouse models of SCC and BCC. Most importantly, in humans the interruption of COX-2 signaling is an effective strategy to treat and chemo-prevent non-melanoma skin cancer in individuals who are at high risk for the disease. However, any potential beneficial effect of this medicine has to be balanced against the adverse effects that are known to be associated with these drugs in a subset of patients.

Prevention of K-Ras- and Pten-mediated intravaginal tumors by treatment with camptothecin-loaded PLGA nanoparticles.

Primary squamous cell carcinoma of the vagina is an uncommon disease that often exhibits few symptoms before reaching an advanced stage. Topical intravaginal therapies for resolving precancerous and cancerous vaginal lesions have the potential to be non-invasive and safer alternatives to existing treatment options. Two factors limit the testing of this approach: lack of a preclinical intravaginal tumor model and absence of safe and effective topical delivery systems. In this study, we present both an inducible genetic model of vaginal squamous cell carcinoma in mice and a novel topical delivery system. Tumors were generated via activation of oncogenic K-Ras and inactivation of tumor suppressor Pten in LSL-K-RasG12D/+PtenloxP/loxP mice. This was accomplished by exposing the vaginal epithelium to a recombinant adenoviral vector expressing Cre recombinase (AdCre). As early as 3 weeks after AdCre exposure exophytic masses protruding from the vagina were observed; these were confirmed to be squamous cell carcinoma by histology. We utilized this model to investigate an anticancer therapy based on poly(lactic-co-glycolic acid) (PLGA) nanoparticles loaded with camptothecin (CPT); our earlier work has shown that PLGA nanoparticles can penetrate the vaginal epithelium and provide sustained CPT release. Particles were lavaged into the vaginal cavity of AdCre-infected mice. None of the mice receiving CPT nanoparticles developed tumors. These results demonstrate a novel topical strategy to resolve precancerous and cancerous lesions in the female reproductive tract.

Effects of heme oxygenase-1 on induction and development of chemically induced squamous cell carcinoma in mice

Heme oxygenase-1 (HO-1) is an antioxidative and cytoprotective enzyme, which may protect neoplastic cells against anticancer therapies, thereby promoting the progression of growing tumors. Our aim was to investigate the role of HO-1 in cancer induction. Experiments were performed in HO-1+/+, HO-1+/−, and HO-1−/− mice subjected to chemical induction of squamous cell carcinoma with 7,12-dimethylbenz[a]anthracene and phorbol 12-myristate 13-acetate. Measurements of cytoprotective genes in the livers evidenced systemic oxidative stress in the mice of all the HO-1 genotypes. Carcinogen-induced lesions appeared earlier in HO-1−/− and HO-1+/− than in wild-type animals. They also contained much higher concentrations of vascular endothelial growth factor and keratinocyte chemoattractant, but lower levels of tumor necrosis factor-α and interleukin-12. Furthermore, tumors grew much larger in HO-1 knockouts than in the other groups, which was accompanied by an increased rate of animal mortality. However, pathomorphological analysis indicated that HO-1−/− lesions were mainly large but benign papillomas. In contrast, in mice expressing HO-1, most lesions displayed dysplastic features and developed to invasive carcinoma. Thus, HO-1 may protect healthy tissues against carcinogen-induced injury, but in already growing tumors it seems to favor their progression toward more malignant forms.

Chemopreventive effects of wild carrot oil against 7,12-dimethyl benz(a)anthracene-induced squamous cell carcinoma in mice

Context: Daucus carota L. ssp. carota (Apiacea) is widely distributed throughout the world and has many uses in traditional medicine.Objective: The present study investigates the chemopreventive effects of oil extract of D. carota umbels on 7,12-dimethyl benz(a)anthracene (DMBA)-induced skin cancer in mice.Materials and methods: D. carota oil extract (DCOE) was prepared by extracting the dried umbels with 50:50 acetone:methanol. Skin papilloma were initiated by DMBA and promoted by 12-O-tetradecanoyl phorobol-13-acetate (TPA). The extract was administered to animals via gavage (0.02 mL of 100% oil), intraperitoneal (0.3 mL of 2% oil), and topical (0.2 mL of 5, 50, and 100% oil) routes for 20 weeks. Tumor appearance, incidence, yield, and volume were compared with those of a non-treated control group.Results: Topical 100% treatment delayed tumor appearance, and inhibited tumor incidence and yield by 40 and 89%, respectively. Topical 50% treatment inhibited tumor incidence and yield by 30 and 83%, respectively, whereas the 5% treatment inhibited tumor yield by 36%. Tumor volume was decreased by 99, 91, and 70% following topical treatments with 100, 50, and 5% oil, respectively. Intraperitoneal treatment inhibited tumor yield by 43%, and decreased tumor volume by 85%, whereas gavage treatment showed minimal effects on both. Intraperitoneal and topical treatment decreased infiltration and hyperplasia with an increase in the level of hyperkeratosis.Conclusion: These findings demonstrate that DCOE has remarkable antitumor activity against DMBA-induced skin cancer compared with non-treated animals paving the ground for further investigations.

E6 and E7 from Beta Hpv38 Cooperate with Ultraviolet Light in the Development of Actinic Keratosis-Like Lesions and Squamous Cell Carcinoma in Mice

Cutaneous beta human papillomavirus (HPV) types appear to be involved in the development of non-melanoma skin cancer (NMSC); however, it is not entirely clear whether they play a direct role. We have previously shown that E6 and E7 oncoproteins from the beta HPV type 38 display transforming activities in several experimental models. To evaluate the possible contribution of HPV38 in a proliferative tissue compartment during carcinogenesis, we generated a new transgenic mouse model (Tg) where HPV38 E6 and E7 are expressed in the undifferentiated basal layer of epithelia under the control of the Keratin 14 (K14) promoter. Viral oncogene expression led to increased cellular proliferation in the epidermis of the Tg animals in comparison to the wild-type littermates. Although no spontaneous formation of tumours was observed during the lifespan of the K14 HPV38 E6/E7-Tg mice, they were highly susceptible to 7,12-dimethylbenz(a)anthracene (DMBA)/12-0-tetradecanoylphorbol-13-acetate (TPA) two-stage chemical carcinogenesis. In addition, when animals were exposed to ultraviolet light (UV) irradiation, we observed that accumulation of p21WAF1 and cell-cycle arrest were significantly alleviated in the skin of Tg mice as compared to wild-type controls. Most importantly, chronic UV irradiation of Tg mice induced the development of actinic keratosis-like lesions, which are considered in humans as precursors of squamous cell carcinomas (SCC), and subsequently of SCC in a significant proportion of the animals. In contrast, wild-type animals subjected to identical treatments did not develop any type of skin lesions. Thus, the oncoproteins E6 and E7 from beta HPV38 significantly contribute to SCC development in the skin rendering keratinocytes more susceptible to UV-induced carcinogenesis.

A novel fusogenic herpes simplex virus for oncolytic virotherapy of squamous cell carcinoma

BackgroundR849 is a neurovirulent γ134.5 gene-deficient form of herpes simplex virus type 1 (HSV-1) and has LacZ genes at the deleted sites of the γ134.5 gene. HF is a spontaneously occurring, fusogenic HSV-1 strain. The purpose of this work was to generate a virus that has the syncytial character of HF, while preserving the γ134.5 gene inactivation profile of R849 virus.ResultsVero cells were infected with R849 and HF simultaneously and two viruses, RH1 and RH2, expressing the LacZ gene and inducing extensive cell fusion were selected. A polymerase chain reaction (PCR)-based analysis suggested that one copy of the γ134.5 gene is lost in RH1, whereas both copies are lost in RH2, and that the γ134.5 gene is replaced by a R849-derived DNA fragment with the LacZ gene. These viruses produced larger plaques and more progeny than the parental viruses. Infection with RH2 decreased the viability of oral squamous cell carcinoma (SCC) cells most strongly. When RH2 was injected into xenografts of oral SCC in nude mice, multinucleated cells were produced and the growth of the tumors was suppressed significantly.ConclusionThese results indicate that novel oncolytic HSV-1 vectors can be produced with the genetic background of the oncolytic HSV-1 HF, and that RH2 is deficient in γ134.5 genes and shows extensive cytopathic effects in oral SCC cells. RH2 may be useful in oncolytic virotherapy for oral SCC.

Intravenous injection of irradiated tumor cell vaccine carrying oncolytic adenovirus suppressed the growth of multiple lung tumors in a mouse squamous cell carcinoma model

Although cancer therapy using replication-selective oncolytic adenoviruses has been available for many years, its anti-tumor efficacy is suboptimal as a result of low and nonspecific infectivity that depends on coxsackie adenovirus receptor expression of the target cancer and normal cells, and generation of an anti-adenovirus neutralizing antibody. In addition, concerns of triggering a severe innate immune response against the adenovirus limit the systemic administration. We developed the carrier cell-based oncolytic virus system (CBOVS) using irradiated tumor cells as carrier cells and concealing the adenovirus (Ad-IAI.3B) inside to improve the specific infectivity. We investigated the anti-tumor effect of CBOVS in a multiple lung tumor mouse model. The ability of CBOVS to infect Ad-IAI.3B to the target cancer cells was examined in vitro in the presence of anti-adenovirus antibodies. To evaluate the systemic effect of CBOVS, we intravenously injected CBOVS into mice with lung tumors (KLN205 cell lines). CBOVS enhanced the infectivity of Ad-IAI.3B to tumor cells in the presence of anti-adenovirus antibodies in vitro. Intravenous injections of CBOVS produced an accumulation of the adenovirus in the lung-bearing tumors and produced a strong anti-tumor effect in vivo. Furthermore, lymphocytes collected from the CBOVS-treated mice induced an increase in cytokines related to the Th1 response (interferon-γ, interleukin-12) by pulsing with KLN205. These findings suggest that CBOVS could protect adenoviruses from neutralizing antibodies and systemically deliver them to lung tumors. Furthermore, CBOVS appears to have potential as a tumor cell vaccine that activates cytotoxic immunity against cancer cells.

Beyond Antiangiogenesis: Intratumorally Injected Bevacizumab Plays a Cisplatin-Sensitizing Role in Squamous Cell Carcinomas in Mice

Background: The anticancer mechanism of bevacizumab beyond antiangiogenesis remains unclear. Here, we investigated whether intratumorally injected bevacizumab could serve as an effective cisplatin sensitizer in squamous cell carcinoma (SCC) in vivo. Methods: Hela and SCC-VII experimental SCC models were established to investigate the anticancer effect of bevacizumab plus cisplatin and the underlying mechanism using immunostaining, TUNEL, and Western blot assays. Results: Bevacizumab-cisplatin therapy markedly inhibited tumor growth and significantly increased survival in both Hela- and SCC-VII-bearing mice compared with single-agent treatments and the untreated control, respectively. Immunostaining of CD34 showed that intratumorally injected bevacizumab significantly reduced microvessel density in bevacizumab-cisplatin and bevacizumab-alone groups of Hela xenografts. TUNEL assay showed that bevacizumab-cisplatin significantly promoted tumor cell apoptosis compared with single-agent treatments and untreated controls in these 2 models. Western blot showed that upregulation of cleaved caspase-3 and downregulation of Bcl-2 and p-Erk expressions are part of the molecular mechanisms beyond angiogenesis which contribute to the cooperative effect of bevacizumab plus cisplatin in the 2 SCC models. Conclusions: Bevacizumab functions not only as an angiogenesis inhibitor but also as a chemosensitizer which enhances the cytotoxicity of cisplatin and promotes apoptosis of SCC cells.

Abstract 4313: Preclinical safety and efficacy of VSV-IFN-b in a syngeneic, immunocompetent model of head and neck squamous cell carcinoma

Abstract Objective& Background: VSV, an RNA virus of the rhabdoviridae family, induces potent cytolytic effects in immortalized cells and cells with defects pathways of IFN or PKR in vitro & in vivo. To take advantage of defects in IFN response in tumors, and to improve the virus’ therapeutic window, “second generation” VSVs expressing IFN-b were developed and characterized (Obuchi et. al. 2003). The aims of this study are to evaluate the effects of VSV-IFN-b in human and rodent head & neck cancer cell lines and to assess the safety and in vivo antitumor efficacy of VSV-IFN-b in syngeneic rat head & neck cancer models. Methods: Rat (FAT-7) and mouse (SCC VII) squamous cell carcinoma cells were infected in vitro with VSV-rat-IFN-b and VSV-mouse -IFN-b. Viruses were propagated using the BHK-21 cells according to established protocols. ELISA & Western blotting were performed to identify the expression of rat-IFN-b post infection, in SCC-VII & FAT-7 cells, respectively. Viral cytotoxicity during normoxia and hypoxia, viral replication and IFN-b expression were assessed to determine sensitivity of rat and mouse SCC cells to VSV-IFN-b. An in vivo model of rat squamous cell carcinoma was established by injection of 3×106/ml of FAT-7 cells SC on the floor of the mouth in immunocompetent female Fisher-344 rats. VSV-rat-IFN-b was administered intratumorally at different doses and schedules when tumors reached a diameter of 70-100 mm3. Antitumor efficacy and toxicity were evaluated during the study. Results: VSV-IFN-b induced significant cytotoxicity and successfully propagated in FAT-7 and SCC VII during normoxia and hypoxia. IFN-b production was increased at 24 and 48 hours after viral infection. Rat SCC cells were more susceptible to VSV-rat-IFN-b than mouse cells, and the cytotoxic effects were similar to VSV-human IFN b on rat cells. Tumors were successfully induced and were palpable at 21 days after inoculation. IT administration of VSV-rat-IFN-b resulted in reduction in tumor size and improved survival compared to the non-treated controls. The antitumor effects of one dose of the virus at 5×108 pfu was more effective than higher doses (5×109) or repeated doses. IV administration of a single dose of VSV-rat-IFN-b (5×108) exerted similar antitumor effects as IT administration. No signs of acute toxicity were observed in treated rats after single or repeated IT or IV virus administration. Conclusion: VSV-IFN-b induces cytotoxicity in FAT-7 and SCC-VII cancer cells during normoxia and hypoxia. In vivo, VSV-rat-IFN-b was associated with antitumor effects and prolongation of survival compared to untreated controls. Characterization of viremia, tissue biodistribution, in vivo viral replication and gene expression in vivo are underway in preparation for future clinical evaluation of VSV-human-IFN-b in subjects with advanced head and neck cancer. 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 4313. doi:10.1158/1538-7445.AM2011-4313

Abstract 2679: Gold nanoparticles enhance the radiation therapy of a murine squamous cell carcinoma

Abstract The purpose of this study is to test the hypothesis that gold nanoparticle (AuNP, nanogold)-enhanced radiation therapy (nanogold radiation therapy, NRT) is efficacious when treating the radiation resistant and highly aggressive mouse head and neck squamous cell carcinoma model, SCCVII, and to identify parameters influencing the efficacy of NRT. Subcutaneous (sc) SCCVII leg tumors in mice were irradiated with x-rays at the Brookhaven National Laboratory (BNL) National Synchrotron Light Source (NSLS) with and without prior intravenous (iv) administration of AuNPs. Variables studied included radiation dose, beam energy, temporal fractionation and hyperthermia. AuNP-mediated NRT was shown to be effective for the sc SCCVII model. AuNPs were more effective at 42 Gy than at 30 Gy (both at 68 keV median beam energy) compared to controls without gold. Similarly, at 157 keV median beam energy, 50.6 Gy NRT was more effective than 44 Gy NRT. At the same radiation dose (∼42 Gy), 68 keV was more effective than 157 keV. Hyperthermia and radiation therapy (RT) were synergistic and AuNPs enhanced this synergy, thereby further reducing TCD50 s (tumor control dose 50%) and increasing long-term survivals. It is concluded that gold nanoparticles enhance the radiation therapy of a radioresistant mouse squamous cell carcinoma. The data show that radiation dose, energy and hyperthermia influence efficacy and better define the potential utility of gold nanoparticles for cancer x-ray therapy. 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 2679. doi:10.1158/1538-7445.AM2011-2679

Solanum incanum extract (SR-T100) induces human cutaneous squamous cell carcinoma apoptosis through modulating tumor necrosis factor receptor signaling pathway

The Solanum species herbs have been used to treat cancer for centuries; however, the underlying mechanisms and effectiveness in vivo remain unclear. SR-T100, extracted from the Solanum incanum, contains solamargine alkaloid as the main active ingredient. Here, we investigated the apoptosis-inducing effects of SR-T100 for targeting squamous cell carcinoma (SCC) in vitro and in vivo. We elucidated the mechanism by which SR-T100 induces apoptosis of human SCCs (A431, SCC4, SCC9, and SCC25) cells. The efficacy and safety issues were addressed regarding topical treatment of SR-T100 on UVB-induced cutaneous SCC of hairless mice and actinic keratoses (AKs) of human. SR-T100 induces apoptosis in human SCCs cell lines by up-regulating the expressions of tumor necrosis factor receptors (TNFRs) and Fas, and downstream adaptors FADD/TRADD of the TNF-α and Fas ligand signaling cascades. SR-T100 also triggered the mitochondrial apoptotic pathway, as up-regulated cytochrome c and Bax, down-regulated Bcl-X(L). Animal experiments showed that all papillomas (35/35) and 27 of 30 UVB-induced microinvasive SCCs in hairless mice disappeared within 10 weeks after once-daily application of topical SR-T100. Furthermore, 13 patients, who suffered with 14 AKs, were treated with once-daily topical SR-T100 gel and 10 AKs cured after 16 weeks, showing negligible discomforts. Our studies indicate that SR-T100 induces apoptosis of SCC cells via death receptors and the mitochondrial death pathway. The high efficacy of SR-T100 in our preclinical trial suggests that SR-T100 is a highly promising herb for AKs and related disorders.

New Dietary Supplements from Medicinal Mushrooms: Dr Myko San—A Registration Report

In December 2010 the Ministry of Health and Social Welfare of the Republic of Croatia registered tablet preparations AGARIKON. 1 and MYKOPROTECT. 1, developed by Dr Myko San-Health From Mushrooms Co., as dietary supplements. This may be the first time for a European manufacturer to successfully register its own medicinal mushroom products in a European country. As a product with a very broad spectrum of action, officially described as a preparation for immunity strengthening and general health improvement, AGARIKON.1 is a result of 20 years of research and practice, and is based on the formulation that has achieved the best tumor growth inhibition rates-above 90% on tumor cell lines of mouse squamous cell carcinoma and fibrosarcoma. Since the usage of massive dosages of proprietary blended liquid mushroom extracts in patients with breast, colorectal, lung, and other cancers significantly improved their survival rates, alleviated side effects of standard oncological therapies, improved their quality of life, and resulted in life prolongation-the very idea is that scientifically verified medicinal mushroom products can be used as powerful biological weapons to fight human malignancies. If progressive modem medicine were redefined in a more effective and humane way, cancer mycotherapy should be a part of a broad concept of biological prevention and therapy of cancer. Also, with a very broad spectrum of action, generally formulated as "to strengthen immunity," MYKOPROTECT. 1 is intended as an important element in the prevention and fighting of serious viral infections, whether they are caused by well-known viruses (hepatitis, HIV, etc.) or newly emerging ones.

Enhanced tumor cures after Foscan photodynamic therapy combined with the ceramide analog LCL29. Evidence from mouse squamous cell carcinomas for sphingolipids as biomarkers of treatment response

To improve anticancer therapeutic success of photodynamic therapy (PDT), combination treatments represent a viable strategy. Sphingolipid analogs combined with anticancer drugs can enhance tumor response. We have shown that LCL29, a C6-pyridinium ceramide, promotes therapeutic efficacy of Photofrin-PDT in mouse SCCVII squamous cell carcinoma tumors. The long-term effect of the combination PDT + LCL29 is unknown. In this study we used the same model to test the long-term curative potential of Foscan-PDT + LCL29. We show that treatment of SCCVII tumors with the combination led to enhanced long-term tumor cure compared to PDT alone. LCL29 itself did not prevent tumor growth. All treatments triggered early increases in tumor-associated C16-ceramide, C18-ceramide, dihydrosphingosine, and global levels of dihydroceramides. PDT-evoked increases in tumor-associated sphingosine-1-phosphate and dihydrosphingosine-1-phosphate remained elevated or were attenuated after the combination, respectively; in contrast, LCL29 had no effect on these two sphingolipids. Our data demonstrate that adjuvant LCL29 improves PDT long-term therapeutic efficacy, implying translational potential of the combination. Furthermore, our findings indicate that changes in the sphingolipid profile might serve as predictive biomarkers of tumor response to treatments.

Investigation of the efficacy of a bevacizumab-cetuximab-cisplatin regimen in treating head and neck squamous cell carcinoma in mice

The efficacy of bevacizumab plus cetuximab-based chemotherapy remains unclear in head and neck squamous cell carcinoma (HNSCC). The aim of this study was to investigate the anticancer efficacy of a bevacizumab-cetuximab-cisplatin triple-agent combination in treating mouse HNSCC. SCC-VII tumor-bearing C3H mice were treated with bevacizumab, cetuximab and cisplatin either alone or in various combinations. In vivo results showed that the largest delay in tumor growth and the maximum increase in survival were not in the triple-agent combination therapy, but in the bevacizumab plus cisplatin therapy. TUNEL assay showed that the apoptosis indices in bevacizumab plus cisplatin group and a triple-agent combination group were 31.6 ± 12.0% and 6.9 ± 1.3%, respectively. Western blot showed that down-regulation of Bcl-2 and up-regulation of cleaved caspase-3 contributed to the anticancer effect of a triple-agent combination and bevacizumab plus cisplatin. Moreover, the maximum level of cleaved caspase-3 expression was not found in the triple-agent combination group but in the bevacizumab plus cisplatin group. Bevacizumab plus cisplatin therapy is better than bevacizumab-cetuximab-cisplatin triple therapy in the mouse HNSCC model. Our findings suggest that the bevacizumab-cetuximab-cisplatin regimen may not be suitable for treating HNSCC.