Pergamon zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA Oral Owl, Eur J Cancer, Vol. 3lB, No. I, pp. 32-36, 1995 Copyright it‘ 1995 Elsevm Science Ltd Printed m Great Britain. All rights reserved 096619551’95 $9.50+0.00 0964-1955(94)00041-7 zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJI Cytokinetic Studies of Oral Cancer Cells Using Bromodeoxyuridine Labelling in Relation to Factors Influencing Prognosis D. Mukhopadhyay, R. Chatterjee and R.N. Chakraborty Cytokinetics of 27 untreated oral squamous cell carcinomas were evaluated by in vitro bromodeox- yuridine (BrdU) labelling. A mononuclear cell suspension was prepared for the labelling using collagenase and DNase treatment. The labelled cells were visualised by immunofluorescence staining with an anti-BrdU monoclonal antibody. The labelling index (LI%) was calculated by determining the percentage of BrdU labelled cells. The LIs of the carcinomas ranged from 3.8 to 19.2%. The LIs of poorly-differentiated (HG3) tumours were 2-3-fold higher compared with those of well-differentiated (HGl) and moderately-differentiated (HG2) carcinomas. Results also showed statistically significant (P< 0.0005) increases in LIs from nuclear grade 1 (NGl) to nuclear grade 3 (NG3) carcinomas. Higher LIs were observed in stage III/IV (10.82 f4.62; 10.36 k4.90) than those in stage I (6.87 f2.09) and II (7.14f 1.87) carcinomas. A significant (P<O.O005) difference in LI values was found between the patients with positive and negative lymph nodes. Good correlation (r= 0.77) was exhibited between the LI values and mitotic counts (MC) of the specimens. These results on oral cancer cell proliferation seem to have prognostic implications. Keywords: HPV, carcinoma, squamous cell, bromodeoxyuridine, oral cancer, cell division, neoplastic zyxwvutsr Oral Oncol, Eur J Cancer, Vol. 31B, No. 1, pp. 32-36, 1995. INTRODUCTION SEVERAL REPORTS [l-3] have shown that the study of human tumour cell kinetics may be of value in understanding neoplastic growth. A knowledge of the cell proliferative status of tumours is indicative of their aggressiveness, sensitivity to antineoplastic agents and is also useful for prognosis [4-71. Tumour cell kinetics are well studied by a recently developed method using in viva [6, 8-111 and in vitro [3, 4, 121 bromodeoxyuridine (BrdU) labelling of cells. BrdU, a thymi- dine analogue which is incorporated into DNA synthesising cells (S-phase) can be detected using a monoclonal antibody against BrdU [ 13, 141. The non-radioactive BrdU method of analysing cell kinetics has been found to be not only faster than the autoradiographic procedure [ 15, 161 but also equally competent, as the two techniques showed good correlation [ 171. This technique has been followed extensively for cytokinetic investigations of brain tumours [4, 181, cervical tumours [19], pulmonary adenocarcinomas [20], tumours of head and neck [8, 211, lung cancer [22], hypopharyngeal and oropharyngeal cancer [9, 231, breast cancer [24], squamous cell carcinomas of oral cavity [ 121 and many other types of tumours [ 10, 11, 251. Correspondence to R. Chatterjee. D. Mukhopadhyay and R. Chatterjee are at the Department of Tumour Virology; R.N. Chakraborty is at the Department of Pathology, Chittaranjan National Cancer Institute, 37 S.P. Mukher- jee Road, Calcutta 700 026, India. Manuscript received 26 Apr. 1994; provisionally accepted 7 June 1994; revised manuscript received 10 Oct. 1994. In this paper, we describe assessment of the presence of cells in S-phase in oral cancer lesions with different histological grading. BrdU incorporation in vitro was monitored using immunofluorescence. The proliferative activity was analysed in relation to other factors, namely clinical stage, nuclear grade and mitotic activity, that are important for prognosis. In the present study we sought to evaluate the relationship between tumour cell proliferation of squamous cell carcinomas of the oral cavity and the factors influencing prognosis. Patients PATIENTS AND METHODS Out of the 49 oral cancer patients treated at the Chittaranjan Cancer Hospital, Calcutta, between June 1992 and May 1993, 27 were randomly selected for this investigation with squa- mous cell carcinomas at different sites of oral cavity (six cases of carcinoma of the left and four of the right cheek; two cases of carcinoma involving the jaw and left cheek; five of lower alveolar carcinoma; three cases of tongue cancer in the lateral margin and one at the lower surface of tongue; three with cancer of the hard palate; two with cancer of the floor of the mouth; one carcinoma of the left jaw). The specimens studied were obtained either surgically or by punch biopsy from the patients prior to the start of their treatment. The age of the patients (23 men and 4 women) ranged from 35 to 65 years (average age, 52 years). 32