MNNG-transformed Bloom syndrome B-lymphoblastoids for the detection of Hodgkin's lymphoma-associated antigen in 2D Westerns.
Twenty-four hour MNNG-exposed Bloom syndrome (BS) B-lymphoblastoid cells with the potential to form single cell colonies in soft agar and nude mouse tumour (2/6 (33%) showed a simultaneous increase in the Ras-expressing cells (using monoclonal antibody to p21 transforming protein) from 20% (at 24 h) to 85% (on day 30). In contrast, there was an absence of Ras-positive cells in MNNG-exposed fresh lymphocytes (PBMCs) from a healthy subject and a presence of only 11-18% of Ras-positive cells in normal (GA3) and unexposed BS B-lymphoblastoid cells. The Western blot analysis using sera samples from Hodgkin's lymphoma patients showed the presence of proteins of 102 and 68 kDa which in 2D Westerns were observed to be unique to BS-MNNG cells with approximate pIs of 5.3 and 5.7, respectively. It is proposed that BS-MNNG cells provide an interesting in vitro human cell model to generate unique cancer-associated antigen(s) in addition to using this system to understand the primary events associated with neoplastic transformation.
Bhalla
A
A
Human Genetics Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
Bamezai
R
R
D000951:Antigens, Neoplasm / Q000302:isolation & purification*; D001402:B-Lymphocytes; D001816:Bloom Syndrome / Q000276:immunology*; D015153:Blotting, Western; D002273:Carcinogens; D002461:Cell Line, Transformed; D006689:Hodgkin Disease / Q000276:immunology*; D006801:Humans; D008769:Methylnitronitrosoguanidine; D016283:Proto-Oncogene Proteins p21(ras) / Q000032:analysis
D000951:Antigens, Neoplasm; D002273:Carcinogens; D008769:Methylnitronitrosoguanidine; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)