De novo programmed cell death in oral cancer.
ABSTRACT The importance of programmed cell death or apoptosis in the maintenance of tissue homoeostasis and the pathogenesis of oral cancer was analysed in relation to apoptosis regulatory proteins, tissue proliferation and tumour histology.
The extent of apoptosis was defined by morphological criteria and the TUNEL (terminal deoxy nucleotidyl transferase-mediated dUTP biotin nick end labelling) assay. p53, bax, bcl-2 and cyclin D1 expression was evaluated by immunocytochemistry. The presence of mutant p53 was analysed using a mutant p53-specific ELISA. An inverse correlation was observed between TUNEL reactivity and histology of the lesion (r = -0.555, P = 0.0001). There was also correlation between TUNEL reactivity and immunoreactivity of apoptosis regulatory proteins. p53 (r = 0.641, P = 0.00023), bcl-2 (r = -0.642, P = 0.00014) and bax (r = 0.651, P = 0.00002). The presence of mutant p53 protein showed an inverse correlation to the extent of apoptosis (r = - 0.301, P = 0.00063). Significant correlation was evident between the bax/bcl-2 ratio and TUNEL (r = 0.652, P = 0.00001) as well as between cyclin D1 and TUNEL reactivity (r = 0.577, P = 0.00001).
Results from this study suggest that apoptosis decreases as histological abnormality increases. Apoptotic regulatory proteins are also altered in a histologically dependent manner. Deregulated proliferation occurs simultaneously with decreased apoptosis during tumour progression in the oral mucosa.
- SourceAvailable from: Zvi Laster[Show abstract] [Hide abstract]
ABSTRACT: Biological markers are necessary for predicting prognosis of salivary malignancies and better understanding the pathogenesis of salivary cancer. We analysed terminal deoxynucleotidyl transferase (TdT)-mediated biotinylated deoxyuridine-triphosphate (dUTP)-biotin nick-end labelling (TUNEL), p53 and Ki67 expression in 66 patients with malignant salivary tumours by immonohistochemistry, and correlated the data with survival, disease-free survival, tumour grade, stage, and local and distant metastasis. TUNEL efficiently predicted poor prognosis in salivary malignancies. The 5-year (5Y) survival probability dropped significantly with the level of TUNEL staining (from 83% in negatively stained tumours to 57 and 24% in TUNEL positively stained levels 1 and 2, respectively), (P=0.042). Extensive Ki67 staining (in addition to TUNEL) reduced the 5Y-survival rate even further and addition of positively stained p53 dropped the 5Y-survival rate to 0. The correlation rates between TUNEL and Ki67 was 58% (P=0.0001), and between TUNEL and p53 it was 50% (P=0.035). Concurrently, TUNEL correlated with metastasis, extracapsular spread, grade and stage. The correlation between TUNEL, p53 and Ki67 staining and survival probabilities, and the pathological grade, stage and metastasis spread of salivary malignancies makes this a highly effective tool in patient follow-up and prognosis.British Journal of Cancer 05/2007; 96(7):1101-6. DOI:10.1038/sj.bjc.6603655 · 4.82 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: Premalignant/potentially malignant-oral lesions and conditions such as oral submucous fibrosis are known to transform into oral cancer. The malignant transformation is often associated with changes at the genetic level that in turn is reflected by the altered expression of proteins related to cell cycle, proliferation, and apoptosis. To evaluate the expression of p53, Ki67 (MIB), bcl2, and bax in oral submucous fibrosis and oral squamous cell carcinoma. To assess the immunohistochemical expression of p53, Ki67 (MIB), bcl2, and bax in 50 cases of oral submucous fibrosis (OSF) and ten each of normal and oral squamous cell carcinoma (OSCC). The labeling indices (LI) of OSF and OSCC were comparable for p53 and Ki67.The p53 LI ranged from 7.9 to 71.9 in OSF and 65.2 to 85.9 in OSCC, and for Ki67 it ranged from 4.39 to 43.23, 18.35 to 42.33, respectively. The p53, Ki67, and bax profiles of OSF and OSCC were altered compared to the normal and these markers could be used as surrogate markers of malignant transformation in OSF.Journal of Oral and Maxillofacial Pathology 05/2011; 15(2):148-53. DOI:10.4103/0973-029X.84478
- [Show abstract] [Hide abstract]
ABSTRACT: The Bcl-2 family of proteins regulates a late step in the apoptosis pathway. Bcl-2 protein is believed to be involved in imparting resistance to programmed cell death or apoptosis induced by chemotherapeutic agents and radiation. The anti-apoptotic function of the Bcl-2 protein appears to be modulated by its ability to heterodimerize with other members of the gene family, predominantly Bax, a protein favouring induction of apoptosis. Susceptibility to undergoing apoptosis may, therefore, be dependent on the ratio between Bcl-2 and Bax. Both Bax and Bcl-2 are regulated by the tumour-suppressor protein p53. The present study therefore aims to study the significance of the Bcl-2:Bax ratio, p53 expression and apoptosis in paediatric acute lymphoblastic leukaemia (ALL). Expression of Bax, Bcl-2 and p53 was determined by immunocytochemistry, and apoptosis was evaluated by an enzymatic end-labelling technique using biotin-dUTP and further confirmed by annexin binding. The presence of mutant p53 was determined using a mutant-p53-specific enzyme-linked immunosorbent assay (ELISA). A total of 32 cases and 20 controls were evaluated. Bcl-2 was found to be expressed in 22/32 of the ALL cases. Pretreatment (spontaneous) apoptosis was observed in 23/32 cases. The mean pretreatment apoptotic index was 11.34 +/- 2.04% with a median value of 7.5%. There was a positive correlation between apoptosis and Bax expression (r = 0.5044; P = 0.0038). There was good correlation between the immunoreactivity of p53 and detection of mutant p53 by ELISA (r = 0.4605; P = 0.0079). The apoptosis index showed a negative borderline correlation to the expression of Bcl-2 protein (r = -0.3181; P = 0.076). There was an inverse correlation between extent of apoptosis and the presence of mutant p53 protein (r = -0.4732; P = 0.006). p53 protein expression also showed a correlation with both Bcl-2 (r = 0.4647; P = 0.007) and Bax (r = 0.4128; P = 0.018). The Bcl-2/Bax ratio, however, showed no significant correlation with apoptosis (r = -0.3131; P = 0.08) or with p53 expression. No significant association was evident between clinical and laboratory parameters with the Bcl-2/Bax protein expression except lymphadenopathy (r = 0.5774; P = 0.03). However, Bax expression showed a borderline correlation with the immediate tumour response to chemotherapy (r = -0.338; P = 0.0628). These patients are being followed-up to look for any association between clinical outcome, Bcl-2/Bax ratio and apoptosis.Journal of Cancer Research and Clinical Oncology 02/2000; 126(1):62-7. DOI:10.1007/s004320050010 · 3.01 Impact Factor