Autoradiographic and immunohistochemical study on the proliferative kinetics of intestinal metaplasia

Abstract

In order to elucidate the proliferative behavior of the intestinal metaplasia around gastric cancer, the authors used both in vitro tritiated thymidine (3H-thymidine) autoradiography and in vivo bromodeoxyuridine (BrdUrd) immunohistochemistry for labeling the proliferative cells of the normal pyloric glands and metaplastic gastric glands. The results of the methods were comparable: The labeling pattern and the rate of labeling were very similar. In the normal pyloric mucosa, the labeled cells were confined to the isthmus region, indicating that pyloric glandular cells are normally renewed from the isthmus region. On the other hand, a zone of the labeled cells was found in the lower half of the intestinalized mucosa, indicating that cell proliferation took place deep in the mucosa, just like the case of normal intestinal glands. The labeling indices of the pyloric mucosa were 19.4% by autoradiography and 18.0% by immunohistochemistry, and that of the intestinalized gastric glands were 25.2% by autoradiography and 24.2% by immunohistochemistry. In conclusion, both 3H-thymidine autoradiography and BrdUrd immunohistochemistry showed that the proliferative kinetics of the intestinalized gastric glands was similar to that of the normal intestinal glands rather than the pyloric glands, i.e. a lower level of proliferative zone and higher labeling index were present.

Fig. 1

Representative presentation of labeled cells by in vitro ³H-thymidine autoradiography. Black-silver grains indicate the incorporation of ³H-thymidine into the DNA. H.E. X 200.

Fig. 2

Representative presentation of labeled cells by in vivo bromodeoxyuridine immunohistochemistry. Dark brown colored nuclei indicate the incorporation of bromodeoxyuridine into the DNA. Immunoperoxidase X 400.

Fig. 3

A schematic representation of the sites and distribution of labeled cells in a specimen from a 46-year-old male, obtained by in vivo bromodeoxyuridine immunohistochemical method. The site of labeled cells was indicated by the number of nuclei counted from the surface of the gland. The labeling index, the percentage of the labeled cells counted in a total number of over 1000 nuclei in the generative cell zone between the level of the uppermost labeled cells and that of the lowermost cells in each specimen in this case was 19.4% in the normal pyloric glands and 22.0% in the intestinalized glands.

Fig. 4

Autoradiograph of normal pyloric mucosa (from a 62-year-old male) obtained by in vitro incubation with ³H-thymidine for 60 min. Labeled epithelial cells are confined to the region of the isthmus of the pyloric gland. H.E. X 200.

Fig. 5

Autoradiograph of intestinalized mucosa (from a 56-year-old male) obtained by in vitro labeling with ³H-thymidine. A zone of the labeled epithelial cells is seen at the lower level of the mucosa. H.E X 400.

Fig. 6

Photomicrograph of normal pyloric mucosa (from a 46-year-old male) obtained by hematolxylin-eosin stain (6A) and in vivo bromodeoxyuridine immunohistochemical method (6B). Labeled epithelial cells are confined to the isthmus region of the gland (6B). X 100.

Fig. 7

Photomicrograph of intestinalized mucosa (from a 46-year-old male) obtained by hematoxylin-eosin stain (7A) and in vivo bromodeoxyuridine immunohstochemcial method (7B). Labeled cells are seen at the lower level of the mucosa (7B). X 100.