Expression Pattern of the Cancer Stem Cell Marker "Nestin" in Leukoplakia and Oral Squamous Cell Carcinoma.

OBJECTIVE: The aim of the present study was to determine and compare the expression pattern and localization of nestin, in an attempt to explore its role in oral carcinogenesis. METHODS: Western blot and immunohistochemistry analysis were performed to study the expression pattern of nestin in normal mucosa, leukoplakia, and oral squamous cell carcinoma samples. Nestin expression was evaluated in the keratinocytes and blood vessels of all the samples and compared with various clinico-pathological parameters. RESULTS: Nestin expression was increased in samples of leukoplakia and oral squamous cell carcinoma when compared with normal mucosa. Among leukoplakia samples, the expression was increased in cases without dysplasia compared to cases with dysplastic features. In cases of oral squamous cell carcinoma, the expression of nestin was found to be decreased with the loss of differentiation. Neoangiogenesis status determined by nestin expression showed an increasing expression from normal mucosa through leukoplakia, to oral squamous cell carcinoma. CONCLUSION: This study has two major findings: (1) identification of nestin as an effective indicator of neoangiogenesis, and (2) nestin may be used as a marker in predicting the early changes in oral carcinogenesis.

Preventive effect of D-carvone during DMBA induced mouse skin tumorigenesis by modulating xenobiotic metabolism and induction of apoptotic events.

The structural integrity and excellent immune system of the skin makes it a protective covering, inspite of its exposure to hazardous compounds. In the present study, the chemopreventive efficacy of D-carvone was studied in 7, 12-dimethylbenz[a]anthracene (DMBA) induced skin carcinogenesis. DMBA (25 µg in 0.1 m L-1acetone) was used to induce skin cancer in Swiss albino mice. Animals were randomly divided into six groups of six animals in each. Different concentrations of D-carvone (10, 20, 30 mg/kg body weight) were used to assess its anticancer effect. Tumor incidence, tumor volume, tumor burden, histological examination and levels of phase I and phase II detoxification agents were analyzed in experimental animals. Further, expression of p53 and various apoptotic proteins including- Bcl-2, Bax was analyzed using immunohistochemistry and enzymatic expression of apoptotic proteins caspase-3 and caspase-9 was carried out by using ELISA. We observed 100% tumor incidence in DMBA-painted animals and our results showed that D-carvone at 20 mg dose significantly prevents skin carcinogenesis. Our results also showed decreased levels of phase I enzymes (Cyt P450 and-Cyt b5) with increased levels of phase II enzymes (GR, GST and GSH) and increased expression of Bax, caspase-3 and caspase-9 with decreased expression of mutated p53 and Bcl-2 in animals treated with DMBA and D-carvone at 20 mg dose. The results of the present study suggest that D-carvone can be used as a chemopreventive agent against skin cancer, as it induces apoptosis in cancer. However, further studies are warranted to check chemopreventive efficacy of D-carvone on cell proliferation, angiogenesis, and metastasis before going to human trial.

Practical significance of utilizing fine needle aspiration cytology as an adjunct diagnostic aid in the preoperative presumptive diagnosis of ameloblastoma.

BACKGROUND: Cytological reports of ameloblastoma are relatively rare in the literature. Appropriate cytologic diagnosis may play a significant role in its preoperative presumptive diagnosis, especially when incisional biopsy findings are inadequate. AIM: To systematically study the detailed cytomorphologic features of ameloblastoma and to evaluate the role of fine needle aspiration cytology (FNAC) in its preoperative diagnosis. MATERIALS AND METHODS: In this study, FNAC was done on 26 cases of intra-osseous jaw lesion, clinically diagnosed as odontogenic tumor or developmental odontogenic cysts and detailed cytopathological interpretation was carried out and the results were correlated with the corresponding histopathology. RESULTS: Of the 26 cases, 15 were found to be ameloblastoma and sensitivity of FNAC in the diagnosis of ameloblastoma was found to be 86.6%. None of the intra-osseous jaw lesion was false positively diagnosed as ameloblastoma in FNAC and hence the specificity was found to be 100%. CONCLUSION: Presence of cohesive epithelial cell clusters exhibiting smaller basaloid cells with peripherally placed tall columnar cells and occasional large squamous cells either adjoining the basaloid epithelial clusters or in isolated group aids in the specific cytological diagnosis of ameloblastoma and FNAC offers an excellent diagnostic aid that may play a significant role in preoperative presumptive diagnosis of ameloblastoma along with incisional biopsy.

Chlorophyllin abrogates canonical Wnt/ß-catenin signaling and angiogenesis to inhibit the development of DMBA-induced hamster cheek pouch carcinomas.

BACKGROUND: Chlorophyllin, a water soluble semi-synthetic food-grade derivative is reported to exhibit a wide range of beneficial health effects. We investigated the effect of chlorophyllin supplementation on Wnt/ß-catenin and vascular endothelial growth factor (VEGF) signaling in the 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis model. METHODS AND RESULTS: Hamsters were divided into 4 groups. The right buccal pouches of group 1 and 2 hamsters were painted with 0.5 % DMBA for 14 weeks. Group 2 animals received in addition chlorophyllin (4 mg/kg bw) in the diet. Group 3 animals received chlorophyllin alone and group 4 animals served as control. mRNA and protein expression of components of Wnt, VEGF, and PI3K/Akt signaling pathways were analyzed by RT-PCR and Western blot analysis. Dietary chlorophyllin administration suppressed the development of HBP carcinomas by altering the expression of several components of the Wnt/ß-catenin signaling pathway. This was associated with inhibition of angiogenesis as evidenced by decreased expression of the proangiogenic factors HIF-1α, VEGF, and VEGFR2. Chlorophyllin administration also downregulated the expression of histone deacetylases involved in epigenetic regulation of tumor angiogenesis. CONCLUSION: Dietary chlorophyllin that abrogates Wnt/ß-catenin and VEGF signaling by targeting a multitude of key signaling molecules is an attractive candidate for preventing tumor progression.