Natural remedies to combat aberrant hallmark signatures including altered glycosylation in oral carcinoma

Background: Tobacco associated oral cancers remain a major concern in India with higher incidence and mortality making it an Indian-centric burning issue. To combat this dreadful disease, we investigated effects of certain natural compounds on the hallmark signatures including glycosylation transcripts levels in oral carcinoma. Methods: The tongue carcinoma cells- SAS cells were treated with tobacco compounds, natural compounds and Cisplatin. RNA was isolated from the cells and converted to cDNA. RT-qPCR was performed to evaluate expression levels of various genes. Results: The treatment of tobacco compounds resulted in similar pattern of altered makers (ST3GAL1, NEU3, FUT5, FUT6, MMP2, BCL2) as observed in tobacco habituated patients. The treatment of Curcumin resulted in down regulation of FUT8 and MMP2 which are known to have a significant association with disease progression and metastasis. Furthermore, Curcumin treatment also resulted in up regulation of the good prognostic glycosylation transcript marker i.e. FUT3 showing its protective effect against the tumor invasion and metastasis. Butein treatment resulted in the down regulation of the worst prognostic indicators i.e. FUT8 and MMP2 in a dose dependent manner. Piceatannol treatment showed better protective effects via down regulation of the markers related to the aggressive disease progression (ST3GAL2, FUT5, FUT8, MMP2, VEGFC). Conclusion: The study provides novel approach of targeting aberrant hallmark signatures including glycosylation with natural compounds which may open the possibility of promising therapeutic strategies using natural compounds alone or in combination with other conventional therapies to alleviate the present scenario of this dreadful disease in India

Clinical Significance of Telomere Length and Associated Proteins in Oral Cancer

Purpose: Telomere shortening is an important event during carcinogenesis. Although studies suggest role of multiple proteins in telomere length regulation, there is dearth of reports in oral cancer which is a leading malignancy in Asian countries especially in India. Thus the present study was carried out to study these mechanisms and explore the pathways involved in telomere—telomerase regulation and identify possible prognostic markers to understand the biology of oral tumors for better treatment approaches. Methods: Telomere length was determined by Southern Hybridisation method, telomeric repeat binding factor (TRF) 1 and 2 expression was detected by Western blot method and telomerase activation by telomeric repeat amplifi cation protocol. Statistical analysis was done using SPSS (Version 10) software. Results: Significant shortening of telomeres was seen in the tumor tissues as compared to normal tissues. Poor prognosis was observed in the patients with higher telomere length in malignant tissue, higher tumor to normal telomere length ratio (T/N TRF LR). Expression of TRF-2 but not TRF-1 protein was signifi cantly higher in the malignant tissues. We also observed telomerase activation in 75 malignant tissues.Conclusions: Our results reveal signifi cant clinical usefulness of telomere length, T/N TRF LR and telomerase activation in the prognosis of oral cancer patients. TRF-2 overexpression in malignant tissues appears to play an important role in telomere length shortening in oral cancer. Abbreviations: TRF—Terminal restriction fragment; TRF-1—telomeric repeat binding factor-1; TRF-2—telomeric repeat binding factor-2; T/N TRF LR—Tumor/ Normal TRF length ratio

Cysteamine Suppresses Invasion, Metastasis and Prolongs Survival by Inhibiting Matrix Metalloproteinases in a Mouse Model of Human Pancreatic Cancer

Background: Cysteamine, an anti-oxidant aminothiol, is the treatment of choice for nephropathic cystinosis, a rare lysosomal storage disease. Cysteamine is a chemo-sensitization and radioprotection agent and its antitumor effects have been investigated in various tumor cell lines and chemical induced carcinogenesis. Here, we investigated whether cysteamine has anti-tumor and anti-metastatic effects in transplantable human pancreatic cancer, an aggressive metastatic disease. Methodology/Principal Findings: Cysteamine’s anti-invasion effects were studied by matrigel invasion and cell migration assays in 10 pancreatic cancer cell lines. To study mechanism of action, we examined cell viability and matrix metalloproteinases (MMPs) activity in the cysteamine-treated cells. We also examined cysteamine’s anti-metastasis effect in two orthotopic murine models of human pancreatic cancer by measuring peritoneal metastasis and survival of animals. Cysteamine inhibited both migration and invasion of all ten pancreatic cancer cell lines at concentrations (,25 mM) that caused no toxicity to cells. It significantly decreased MMPs activity (IC50 38–460 mM) and zymographic gelatinase activity in a dose dependent manner in vitro and in vivo; while mRNA and protein levels of MMP-9, MMP-12 and MMP-14 were slightly increased using the highest cysteamine concentration. In vivo, cysteamine significantly decreased metastasis in two established pancreatic tumor models, although it did not affect the size of primary tumors. Additionally, cysteamin

Role of aberrant glycosylation enzymes in oral cancer progression

BACKGROUND: Carcinogenesis, a multistep process involves sequential changes during neoplastic transformation. The various hallmarks of cancer aid in cell survival, proliferation, and dissemination. Aberrant glycosylation, a recently defined hallmark of cancer, is influenced by glycosylation enzymes during carcinogenesis. Therefore, the present study measured α-2,3 and α-2,6 sialyltransferase (ST), sialidase, and α-L-fucosidase activity in patients with oral precancerous conditions (OPC) and oral cancer patients. SUBJECTS: The study enrolled 100 oral cancer patients, 50 patients with OPC, 100 healthy controls, and 46 posttreatment follow-ups of oral cancer patients. Blood and saliva were collected from all the participants. MATERIALS AND METHODS: Sialidase activity was measured by spectrofluorimetric method, α-2,3 and α-2,6 ST by ELISA using biotinylated lectins, and α-L-fucosidase by spectrophotometric method. RESULTS: The results depicted increased levels of sialidase, α-2,3 and α-2,6 ST, α-L-fucosidase in patients with OPC and oral cancer patients. Receiver operating characteristic curve indicated significant discriminatory efficacy in distinguishing controls and oral cancer patients for serum and salivary sialidase and α-L-fucosidase activity, and serum α-2,6 ST. Furthermore, serum and salivary α-L-fucosidase activity and serum sialidase activity significantly distinguished controls and patients with OPC. Serum and salivary sialidase, α-L-fucosidase, and serum α-2,3 ST activity were higher in patients with metastasis as compared to nonmetastatic patients. Higher values of serum α-L-fucosidase activity were significantly associated with low-overall survival. CONCLUSION: The increased levels of enzymes correlated with tumor initiation, progression, and metastasis in oral cancer patients. The alterations in glycosyltransferases/glycosidases thus support the view of glycosylation as a hallmark of cancer

Trisomy 8 in leukemia: A GCRI experience

Trisomy of chromosome 8 is frequently reported in myeloid lineage disorders and also detected in lymphoid neoplasms as well as solid tumors suggesting its role in neoplastic progression in general. It is likely to be a disease-modulating secondary event with underlying cryptic aberrations as it has been frequently reported in addition to known abnormalities contributing to clinical heterogeneity and modifying prognosis. Here, we share our findings of trisomy 8 in leukemia patients referred for diagnostic and prognostic cytogenetic assessment. Total 60 cases of trisomy 8, as a sole anomaly or in addition to other chromosomal aberrations, were reported (January 2005-September 2008). Unstimulated bone marrow or blood samples were cultured, followed by GTG banding and karyotyping as per the ISCN 2005. Patients with +8 were chronic myeloid leukemia (CML) (36), acute myeloid leukemia (AML) (17), and acute lymphoblastic leukemia (ALL) (7). In 7 patients, trisomy 8 was the sole anomaly, whereas in 6 patients +8 was in addition to normal clone, in 47 patients, the +8 was in addition to t(9;22), t(15;17), and others, including 3 with tetrasomy 8. Only one patient showed constitutional +8. The present study will form the basis of further cumulative studies to correlate potential differential effects of various karyotypic anomalies on disease progression and survival following a therapeutic regime. To unravel the role of extra 8 chromosome, constitutional chromosomal analysis and uniparental disomy will be considered

Cysteamine inhibited cell migration and invasion of pancreatic cancer cell lines.

<p>A. For the matrigel invasion assay, cells were incubated with increasing concentrations of cysteamine in the matrigel chamber for 24 hours. The number of invaded cells on the opposite side of the membrane was counted. B and C. Wound healing assay. Cells were cultured until confluent and scratched using a sterilized yellow tip. They were incubated for 24 hours with 0–5 mM of cysteamine and the area of the wound between cell layers was measured. The black horizontal lines in figure (vehicle) depict the area at time 0 when wound was made and indicate the area of the wound when cells were scratched (B). D. For cell viability assay, pancreatic cancer cells were incubated with various concentrations of cysteamine and live cell number was counted at 24 and 48 hours. Data are expressed as mean ± S.D. of triplicate determinations. Statistical significances are shown by †: P<0.01 and ‡: P<0.001.</p

Cysteamine inhibited primary tumors and metastatic lesions in an orthotopic pancreatic cancer <i>in vivo</i>.

<p>HS766T and MIA-PaCa2 cells (2×10⁶) were implanted directly into the pancreas of 5–6 week-old female nude <i>nu/nu</i> mice and then peritoneal cavity and skin were closed using clips. Increasing doses of cysteamine were injected s.c. as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0034437#s2" target="_blank">materials and methods</a>. Primary tumors and metastatic lesions of ≥ 5 mm were counted in vehicle and cysteamine treated mice. <i>a</i>: P<0.05, <i>b</i>: P<0.01 and <i>c</i>: P<0.05, <i>d:</i> P<0.001. +: moderate ascites, ++: severe ascites, –: no ascites.</p