Cancer Science & Therapy

Dermal and subdermal metastases as well as primary invasive melanoma allow an excellent opportunity to study the effects of intralesional therapy. The therapy consisted of weekly intralesional administration of nontoxic doses of granulocyte-macrophage colony stimulating factor at 500 microgram, and in case of failure to establish complete tumor response at the injection sites in 4-6 weeks; weekly intralesional interleukin-2 at 18 million international units was substituted. The results revealed complete response in patients with multiple small in transit metastases ranging in size from few millimeters to one centimeter (n=4) for over 31-48 months, but not in any of the large sclerotic lesions of 2 centimeter or more (n=3). Complete responses were elicited for 6 months in one of two patients with distant metastases who had palpable subdermal tumors treated with granulacyte-macrophage colony stimulating factor. A patient with primary cutaneous melanoma with a satellite lesion and and a large regional lymph node metastasis received both cytokines in the skin sites, one week prior to surgery, had complete tumor necrosis, intense immune response at the injection sites and at the regional lymph nodes, with over 36 months disease free survival. This could be an affective new approach to tumor-specific adjuvant immunotherapy.

Background: High-grade gliomas (HGG) are highly infiltrative malignancies, causing considerable mortality in child- and adulthood, necessitating new therapies. Novel therapies directed against multiple epidermal growth factor family (ErbB) members are potentially effective in HGG.

Aim: To assess ErbB family expression in normal brain and pediatric and adult HGG in silico and to determine radiosensitizing property of the pan-ErbB inhibitor CI-1033, in HGG cells in vitro.

Material and methods: In silico mRNA array expression analysis was performed to assess EGFR, ERBB2, ERBB3, ERBB4 gene expression in normal brain, adult and pediatric HGG. ErbB family protein expression was determined in HGG cell lines using Western blot. Sulforhodamine-B assay was used to assess cytotoxicity of CI- 1033 andclonogenic assays to determine radiosensitization. The effect on cell cycle distribution and PI3K-Akt/Ras- MAPK signalling of CI-1033 ± radiation was measured using flow cytometry.

Results: EGFR and ERBB2 were significantly overexpressed in datasets of pediatric and adult HGG. Heterogeneous protein expression of EGFR, ErbB2, 3 and 4 was observed in HGG cell lines. CI-1033 IC50 values of 1.0 μM, 2.5 μM and 4.3 μMwere found in D384MG, U-251 MG and Gli-6 cells, respectively. CI-1033 significantly sensitized Gli-6 and D384MGcells to radiation, with 24 and 48 hrs pre-treatment respectively.

Conclusion: EGFR and ErbB2 are overexpressed in adult and childhood HGG. Irreversible pan-ErbB inhibition by CI-1033 is cytotoxic and radiosensitizes HGG cell lines in vitro, warranting further in vivo studies.

Glioblastoma represents the deadliest tumors of the central nervous systems and their bleak prognosis remains unchanged over the past few decades and requiring new treatments that curb the tumor progression. Cancer cells possess high reactive oxygen species (ROS) levels when compared to their normal control counterparts. We have earlier characterized the ROS-scavenging annexin protein, BjAnn1 from Brassica juncea and demonstrated its peroxidase activity. Since annexins are widely conserved across evolutionary lines with involvement in stress alleviation, we attempted to verify the effect of ectopic expression of ROS-scavenging BjAnn1 in human glioblastoma cell lines. The multiple sequence alignment revealed that BjAnn1 showed homology to human annexin A13 (73.18%), annexin A2 (70.66%), annexin A3 (72.53%) and annexin A8 (70.03%). Stable expression of BjAnn1 in U251 and U87 cells inhibited proliferation, migration and invading abilities in correlation with significant decrease in ROS levels in comparison to the empty vector (EV)-expressing controls. Multiple gene-profiling using cDNAPCR arrays revealed a prominent transcriptional upregulation of oxidative stress responsive genes including CCS, CYB, DUOX2, FOXM, GSS, MBL2, MT3, MTL5, NME5, PXDN, SOD2 and SOD3 in BjAnn1-expressing glioblastoma cells. Western blotting confirmed the significant increase in the expression of antioxidant enzymes SOD2 (MnSOD) and SOD3 (Cu/ZnSOD) in BjAnn1-expressing cells. We also observed a significant inhibition in nuclear translocation of p65 and p50, NF-κB reporter activity and expression of NF-κB-target genes in BjAnn1 expressing cells. In summary, our study shows a prominent tumor-suppressive role of ROS-scavenging BjAnn1 in glioblastoma cell lines suggesting it as a novel candidate for efficient gene therapy in glioblastoma.

Globo H, a hexasaccharide initially identified as a ceramide-linked glycolipid from human breast cancer cell line MCF-7, is over-expressed on the surface of many common cancers, but its function is unknown. Here we demonstrated the uptake of globo H ceramide (GHCer) by human peripheral blood mononuclear cells (PBMC) upon co-culture with MCF-7 cells. Significantly, the expression of globo H on tumor-infiltrating lymphocytes was observed in 61% of globo H positive breast cancer tissues. Addition of synthetic GHCer to human PBMC, mouse splenocytes or purified CD4+ T cells inhibited their proliferative response to anti-CD3/CD28 to 60 ± 1%, 50 ± 7% and 62 ± 5% of control, respectively, and reduced the secretion of IL-2, IFN-γ and IL-4. GHCer also significantly suppressed the proliferation of splenocytes or purified CD19+ B cells to 45 ± 10% and 26 ± 3% of control in response to LPS or LPS + IL4 +CD40 ligand and their IgM production to 12 ± 5% and, 8 ± 3%, and IgG to 34 ± 9%, 35 ± 5%, respectively, with neglible induction of plasma cells. Ceramide displayed no such inhibitory effects. On the other hand, GHCer failed to raise the number of regulatory T cells, or their expression of FOXP3/CTLA4, nor did it increase apoptosis. Notably, GHCer significantly suppressed the Notch1 signaling during activation of human PBMC and murine T and B cells. Furthermore, GHCer upregulated the expression of ID3 and itch by 2.1±0.2 and 4.7 ± 0.4 fold, respectively, leading to ID3-dependent downregulation of Notch1 and itch-mediated Notch1 degradation. These results provide the first evidence for GHCer to facilitate the escape of cancer cells from immune surveillance.

Aim of the study was to determine the effect of essential oil of Wedelia chinensis (Osbeck) on lung metastasis developed by B16F-10 melanoma cell line in C57BL/6 mice. The essential oil extracted by hydro distillation method was evaluated by GC-MS for its bioactive compounds. Male C57BL/6 mice were injected with B16F-10 melanoma cells through tail vein and simultaneously treated with the essential oil of Wedelia chinensis (Osbeck). The selected dose of drug (50 μg of essential oil) showed its cytotoxicity on B16 F-10 melanoma cell line with 65.17% of death within 24hrs in-vitro. We observed a statistically significant increase in the number of apoptotic cells in essential oil treated group when compared with the normal and cancer control groups. We have also observed an increased level of expression of apoptosis inducing molecules such as p53 and caspase-3 in treated group when compared to cancer group as an indicator of utilization of such molecules for inducing apoptosis. The present investigations indicate the effect of essential oil of Wedelia chinensis (Osbeck) in suppressing the metastatic potential of B16F-10 cells in lungs as well as tumor directed angiogenesis. It has shown a powerful apoptotic effect by increasing the levels of p53 and caspase-3 enhancing apoptotic nuclei and thereby it inhibits cancer progress. This drug can be therefore taken as a key therapeutic agent to treat cancer in future with extensive molecular studies.

Bladder Cancer (BCa) is the most common malignancy arising from the urinary tract. One of the mainstays of diagnosis, staging, and therapeutic decision-making for BCa is accurate and appropriate imaging. The ability to identify metastatic disease preoperatively is of utmost importance in determining treatment. Advances in standard cross sectional imaging techniques like Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) have improved imaging of bladder cancer. Over the last decade, ¹⁸F-fluorodeoxyglucose (FDG) Positron Emission Tomography (PET) in combination with CT (¹⁸F-FDG PET/CT) has become an important non-invasive imaging modality for the preoperative staging of various malignancies. ¹⁸F-FDG PET/CT is useful for detection of metastatic disease in BCa, but the ability to detect primary bladder wall lesions remains to be elucidated. To overcome the problem with urinary excretion of ¹⁸F-FDG, new PET tracers are being tested. MRI is an accurate technique for the local staging of BCa due to its superior spatial and contrast resolution. Anatomical MRI has a modest utility in NMstaging of BCa. However, incorporation of functional MR techniques, such as diffusion weighted MRI can improve the results for lesion detection and staging and multi-parametric MRI`s role is yet to be explored widely. The aim of this review is to present the recent advances in PET/CT and MRI in BCa, with particular focus on improvements in staging.