Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Effects of Bacillus Calmette-Guerin (BCG) on the DNA Synthesis of Mouse Duodenal Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells

Bacillus Calmette-Guerin (BCG) 투여가 Ehrlich 종양세포를 이식한 생쥐 샘창자 상피세포의 DNA합성에 미치는 영향

  • Ko, Jeong-Sik (Department of Anatomy, College of Medicine Soonchunhyang University) ;
  • Kim, Heung-No (Department of Anatomy, College of Medicine Soonchunhyang University) ;
  • Park, Kyung-Ho (Department of Anatomy, College of Medicine Soonchunhyang University) ;
  • Park, Dae-Kyoon (Department of Anatomy, College of Medicine Soonchunhyang University) ;
  • Kim, Duk-Soo (Department of Anatomy, College of Medicine Soonchunhyang University)
  • 고정식 (순천향대학교 의과대학 해부학교실) ;
  • 김흥노 (순천향대학교 의과대학 해부학교실) ;
  • 박경호 (순천향대학교 의과대학 해부학교실) ;
  • 박대균 (순천향대학교 의과대학 해부학교실) ;
  • 김덕수 (순천향대학교 의과대학 해부학교실)
  • Received : 2010.10.16
  • Accepted : 2010.12.20
  • Published : 2010.12.31
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Abstract

This experiment was performed to evaluate the morphological responses of the duodenal epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Bacillus Calmette-Guerin (BCG). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8\sim0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of $0.7{\mu}Ci$/g of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and duodenal tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab, England) in a dark room and dried and were placed in a light-tight box. The number of labeled epithelial cells in the duodenal mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On the light microscopic study, duodenal mucosae had no severe morphological changes following the injection of BCG. In the tumor control and BCG treated groups, a number of small lymphocytes and eosinophile leucocytes are slightly increased as compared with those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 632.3 (${\pm}14.47$), 761.7 (${\pm}27.65$) and 505.0 (${\pm}17.09$) respectively. From the above results, BCG may suppress the DNA synthesis of the duodenal epithelial cells, but does not results severe structural defect on the duodenal mucosae. And it is suggested that BCG may greatly improve the anticancer therapy on certain kind of cancer.

이 실험은 Ehrlich 종양세포를 샅부위에 이식한 후 고농도의 BCG(Bacillus Calmette-Guerin)를 피부밑조직에 반복 투여하였을 때, 샘창자점막의 형태학적 변화와 함께 DNA합성지수를 비교 연구하고자 3H-thymidine을 투여한 후 자기방사법적 연구를 시행하였다. 실험동물로는 체중 25 g 내외의 ICR 흰생쥐를 사용하였으며 정상대조군을 제외한 종양대조군과 BCG투여군의 동물에는 샅부위 피부밑조직에 각각 $1{\times}10^7$의 Ehrlich 종양세포를 이식하였다. BCG투여군은 종양세포이식 다음날부터 농축 건조된 BCG를 생리식염수에 용해시킨 다음, 일정량을 하루건너 한 번씩 7회 피부밑조직에 주사하였다. 종양대조군은 종양세포이식 후에 생리식염수를 하루건너 한 번씩 7회 피부밑조직에 주사하였고, 정상대조군은 종양세포를 이식하지 않은 동물을 사용하였으며 각 군에는 5마리씩의 동물을 배정하였다. 자기방사법적 관찰을 위하여는 약제를 마지막으로 주사한 다음날 3H-thymidine $0.7{\mu}Ci$/gm를 꼬리에 한차례 정맥주사하고, 70분 후 도살하여 샘창자조직을 떼어내어 10% 포르마린으로 고정하였다. 관찰은 샘창자조직이 세로로 잘 절단된 창자움 부위를 택하여 점막근육판을 따라 점막길이 3.5mm의 창자샘상피에 분포하는 3H-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylineosin (H-E)염색을 시행하였다. 일반조직관찰에서 종양대조군과 BCG투여군은 정상대조군에 비해 샘창자 점막 고유판에 림프구와 호산성백혈구가 다수 관찰된 것 이외에는 형태적으로 큰 변화를 볼 수 없었다. 자기방사법적 연구에서 정상대조군, 종양대조군 및 BCG투여군은 점막길이 3.5 mm 당 출현하는 표지세포수가 각각 632.3 (${\pm}14.47$), 761.7 (${\pm}27.65$) 및 505.0 (${\pm}17.09$)개였다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 생쥐에 BCG를 반복 투여하면 샘창자 점막상피세포의 DNA합성을 유의하게 억제하면서도 형태적인 변화가 경미하였으므로 BCG는 항암치료시 항암효과를 높일 수 있는 좋은 약제라고 생각된다.

Keywords

References

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