Cell Transformation Assay Using Balb/c 3T3 Cells or Bhas 42 Cells for the Efficient Detection of Tumour Promoters

Abstract

In this paper, we report three interlaboratory validation studies with special interest in detecting tumour promoters. The former two validation studies were on the improved two-stage in vitro transformation assay using Balb/c 3T3 cells. The third one was on the Bhas 42 cell assay. From the results, we could confirm that, on the whole, the improved two-stage transformation assay was sensitive, reproducible and reliable, while the Bhas 42 cell assay was a very sensitive and reproducible method for detecting tumour-promoting potential. However, there was some variation in the sensitivity among the laboratories. In order to cope with this sensitivity variation, further refinement is necessary, which will increase the applicability of the methods.

Keywords

Balb/c 3T3 cells Bhas 42 cells interlaboratory validation study in vitro transformation assay promoters

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References

Tsuchiya

, and 39 co-authors (1999). An interlaboratory validation study of the improved transformation assay employing Balb/c 3T3 cells: results of a collaborative study on the two-stage cell transformation assay by the Non-genotoxic Carcinogen Study Group. ATLA 27, 685–702.

Tsuchiya

, and 35 co-authors (2002). The 2nd validation study in Japan on the modified two-stage transformation assay employing Balb/c 3T3 cells. Poster presented at The Fourth World Congress on Alternatives and Animal Use in the Life Sciences, New Orleans, LA, USA, 11–15 August 2002.

Ohmori

, Asada

, Sasaki

, Tanaka

, & Umeda

(2002). Development of an assay method for predicting tumor promoters using v-Ha-ras-transfected Balb/c 3T3 cells (BHAS 42 cells). In Program and Abstracts, Fourth World Congress on Alternatives and Animal Use in the Life Sciences, New Orleans, LA, USA, p. 268. Alternatives Congress Trust.

IARC/NCI/EPA Working Group (1985). Cellular and molecular mechanisms of cell transformation and standardization of transformation assays of established cell lines for the prediction of carcinogenic chemicals: overview and recommended protocols. Cancer Research 45, 2395–2399.

Tsuchiya

, & Umeda

(1995). Improvement in the efficiency of the in vitro transformation assay method using Balb/c 3T3 cells. Carcinogenesis 16, 1887–1894.

Tsuchiya

, & Umeda

(1997). Relationship between exposure to TPA and appearance of transformed cells in MNNG-initiated transformation of Balb/c 3T3 cells. International Journal of Cancer 73, 271–276.

Sasaki

, Mizusawa

, & Ishidate

(1988). Isolation and characterization of ras-transfected BALB/3T3 clone showing morphological transformation by 12-O-tetradecanoyl-phorbol-13-acetate. Japanese Journal of Cancer Research 79, 921–930.

Nishiyama

, Tsuchiya

, Yoshimura

, & Umeda

(2002). Three-step method of data analysis, incorporating multiple contrast method to detect carcinogenic substances in cell transformation assay. In Program and Abstracts, Fourth World Congress on Alternatives and Animal Use in the Life Sciences, New Orleans, LA, USA, p. 268. Alternatives Congress Trust.