Immunohistochemical study on distribution of proliferating cells and tumor formations in intestinal tracts of 1,2-dimethylhydrazine-treated rats

1,2-dimethylhydrazine을 투여한 rat 장관의 증식세포 분포와 종양발생에 대한 면역조직화학적 연구

  • Kang, Won-hwa (College of Veterinary Medicine Gyeongsang National University) ;
  • Kwak, Soo-dong (Institute of Animal Medicine, Gyeongsang National University)
  • 강원화 (경상대학교 수의과대학) ;
  • 곽수동 (경상대학교 동물의학연구소)
  • Received : 1996.06.17
  • Published : 1997.03.25

Abstract

The present investigation was focussed mainly on the development of the tumors and proliferating cells on the intestinal tracts of 1, 2-dimethyl-hydrazine(DMH)-treated young or adult rats. 26 rats(Wistar, 14 young rats weighting approximately 130~180gm and 12 adult rats weighting approximately 500~550gm) were given subcutaneously once weekly with 20mg of DMH/kg body weight(BW)/week for 8~22 weeks. Individual body weight were recorded weekly at the same day and time. The rats were killed at 8, 13, 15. 17, 19, 21 and 22 weeks. The intestinal tracts were opened longitudinally and carefully examined for tumors. The localization, number, and size of tumors were noted. Tumor-bearing areas were dissected out and fixed on neutral buffered 10% formalin and normal-looking mucosa from 8~22 weeks rats were also taken for fixation. Paraffin sections were stained by H-E for histopathological examination or with immunohistochemical stain for bromodeoxyuridine(Brdur) positive cells. 1. The growth proportion of body weight appeared to be decreased in the DMH-treated young rats than in control young rats and body weight of DMH-treated adult rats appeared to be 13.4% or less lower than weighted on 0 week. 2. Macroscopically, the developed tumors in the intestinal tracts were not observed as early as the 13 weeks after DMH treatment. The number of developed tumors per rat was found to be 14.3, 18.8, 22.3 in 15, 17 and 22 weeks. The numbers of tumors in intestinal regions per rat were 2.1, 4.3, 5.4, 2.5 in duodenum, jejunum, ilium and colon on 15 weeks, 2.3, 6.4, 7.8, 2.3, on 17 weeks, and 2.7, 9.3, 9.0, 1.3 on 22 weeks, respectively and the ileum and jejunum were higher in appearance rate of tumors and tumor types are dome shapes and diameter of largest tumor were 6.3mm. 3. Histopathologically, intestinal mucosa were thickened by the irregular distorted and distended crypts following hyperplasia. The tumors developed on the mucosa and submucosa and were recognized to be adenocarcinoma. 4. Immunohistochemically, the labeling index(LI) was calculated as the ratio of the number of Brdur-labeled cells to the total number of column cells of the crypts with longitudinal axis. LI of Brdur positive cells per crypt were 5.6%, 8.0% on small intestine of control and 22 week group, respectively and 3.7%, 12.7% on large intestine of control and 22 week group, respectively and were appeared to be increase in 22 week group than in control group and to be more number of proliferating cells in 22 week group than in control group. 5. LI of Brdur positive cells in 1, 2, 3, 4, 5 segments of crypt column were 11.7%, 10.7%, 3.8%, 0.6%, 0% in small intestine of control group and 23.5%, 11.8%, 2.3%, 2.4%, 0.8% in small intestine of 22 week group, and 5.4%, 7.4%, 3.8%, 1.0%, 0.4% in large intestine of control group and 29.5%, 20.3%, 5.9%, 6.3%, 1.3% in large intestine of 22 week group respectively. So results indicate that the number of proliferating cells by DMH treatment increase and were concentrated on the 1, 2 segments of crypt columns.