Bacillus Calmette-Guerin (BCG) 투여가 Ehrlich 종양세포를 이식한 생쥐 샘창자 상피세포의 DNA합성에 미치는 영향
Effects of Bacillus Calmette-Guerin (BCG) on the DNA Synthesis of Mouse Duodenal Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
이 실험은 Ehrlich 종양세포를 샅부위에 이식한 후 고농도의BCG(Bacillus Calmette-Guerin)를 피부밑조직에 반복 투여하였을 때, 샘창자점막의 형태학적 변화와 함께 DNA합성지수를 비교 연구하고자 3H-thymidine을 투여한 후 자기방사법적 연구를시행하였다. 실험동물로는 체중 25 g 내외의 ICR 흰생쥐를 사용하였으며정상대조군을 제외한 종양대조군과 BCG투여군의 동물에는 샅부위 피부밑조직에 각각 1×107의 Ehrlich 종양세포를 이식하였다. BCG투여군은 종양세포이식 다음날부터 농축 건조된 BCG 를 생리식염수에 용해시킨 다음, 일정량을 하루건너 한 번씩 7회피부밑조직에 주사하였다. 종양대조군은 종양세포이식 후에 생리식염수를 하루건너 한 번씩 7회 피부밑조직에 주사하였고, 정상대조군은 종양세포를 이식하지 않은 동물을 사용하였으며 각군에는 5마리씩의 동물을 배정하였다. 자기방사법적 관찰을 위하여는 약제를 마지막으로 주사한 다음날 3H-thymidine 0.7 μCi/gm 를 꼬리에 한차례 정맥주사하고, 70분 후 도살하여 샘창자조직을 떼어내어 10% 포르마린으로 고정하였다. 관찰은 샘창자조직이 세로로 잘 절단된 창자움 부위를 택하여 점막근육판을 따라점막길이 3.5mm의 창자샘상피에 분포하는 3H-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylineosin (H-E)염색을 시행하였다. 일반조직관찰에서 종양대조군과 BCG투여군은 정상대조군에비해 샘창자 점막 고유판에 림프구와 호산성백혈구가 다수 관찰된 것 이외에는 형태적으로 큰 변화를 볼 수 없었다. 자기방사법적 연구에서 정상대조군, 종양대조군 및 BCG투여군은 점막길이3.5 mm 당 출현하는 표지세포수가 각각 632.3 (±14.47), 761.7(±27.65) 및 505.0 (±17.09)개였다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 생쥐에BCG를 반복 투여하면 샘창자 점막상피세포의 DNA합성을 유의하게 억제하면서도 형태적인 변화가 경미하였으므로 BCG는항암치료시 항암효과를 높일 수 있는 좋은 약제라고 생각된다.
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 1×107 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.: 0.03×108~0.32×108 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 μCi/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 (±14.47), 761.7 (±27.65) and 505.0 (±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.
참고문헌 (34)
- Aldwell FE, "Oral vaccination of mice with lipidencapsulated Mycobacterium bovis BCG: anatomical sites of bacterial replication and immune activity", Immunol Cell Biol, 83(5), pp.549~553 (2005)
- Barbeito CG, "Mitotic activity of the duodenal crypt enterocytes in mice transplanted with EA21a mammary carcinoma", Cell Biol Int, 26(1), pp.123~125 (2002)
- Beaulieu JF, "Hormonal regulation of epithelial cell proliferation in the fetal mouse duodenum in vitro", Anat Rec, 217(3), pp.250~255 (1987)
- Cameron IL, "Cell proliferation and renewal in the mammalian body, in : Cellular and Molecular Renewal in the Mammalian Body", pp.45~79 (1971), Academic Press
- Chirigos MA, "Immunomodulators: Current and future development and application", Thymus, 19(1), pp.7~20 (1992)
- Chung JW, "Human tissue biology", pp.538~675 (2001), Soonoonsa Publishing
- Ciccocioppo R, "Small bowel enterocyte apoptosis and proliferation are increased in the elderly", Gerontology, 48(4), pp.204~208 (2002)
- Clark WG, "Goth’s medical pharmacology", pp.704~714 (1992), Mosby Year Book
- DeHaven JI, "Antibiotic and steroid therapy of massive systemic Bacillus Calmette-Guerin toxicity", J Urol, 147, pp.738~742 (1992)
- El-Demiry MIM, "Local immune responses after intravesical BCG treatment for carcinoma in situ", Brit J Urol, 60, pp.543~548 (1987)
- Gerlovich ESh, "Proliferative properties of the epithelium of the duodenal mucosa in chronic hepatitis and liver cirrhosis", Arkh Patol, 39(1), pp.43~47 (1977)
- Gilman AG, "Goodman and Gilman’s the pharmacological basis of therapeutics", pp.1240~1306 (1985), Macmillan Pub Co
- Groza P, "The gastrointestinal tract in hypokinetic rats", Life Sci Space Res, 17, pp.199~204 (1979)
- Helpap B, "Repair of gastric ulcer. A cell kinetic study", Virchows Arch, 392, pp.159~170 (1981)
- Holt PR, "Small intestinal crypt cell proliferation rates are increased in senescent rats", J Gerontol, 44(1), pp.9~14 (1989)
- Karam SM, "Dynamics of epithelial cells in the corpus of the mouse stomach. I. Identification of proliferative cell types and pinpointing of the stem cell", Anat Rec, 236, pp.259~279 (1993)
- Klimpel GK, "BCG-induced suppressor cells I. Demonstration of a macrophage-like suppressor cell that inhibits cytotoxic T cell genetation in vitro", J Immunol, 120, pp.563~569 (1978)
- Ko JS, "Effects of antitumor agents on the spleen of mouse implanted with ehrlich carcinoma cells: An autoradiographic study", Korean J Anat, 33(3), pp.315~326 (2000)
- Lamm DL, "BCG in perspective: Advances in the treatment of superficial bladder cancer", Eur Urol, 27(2), pp.2~8 (1995)
- Lamm DL, "Enhanced natural killer (NK) cell activity with BCG and vitamin treatment (abstract 991)", J Urol, 151, pp.475~ (1994)
- Martins MJ, "Effect of fasting on rat duodenal and jejunal microvilli", Clin Nutr, 20(4), pp.325~331 (2001)
- Mathe G, "Immune manipulatioon by BCG administered before or after cyclophosphamide for chemoimmunotherapy of L1210 leukemia", Eur J Cancer, 10, pp.661~670 (1974)
- Popiela T, "Postoperative immunochemotherapy (BCG+5-FU) in advanced gastric cancer", Anticancer Res, 8, pp.1423~1428 (1988)
- Ratliff TL, "Requirement of a thymus dependent immune-response for BCG-mediated antitumor activity", J Urol, 137, pp.155~158 (1987)
- Roitt I, "Immunology", pp.33~44 (1993), Mosby-Year Book
- Rubio CA, "DNA synthesis in the gastroduodenal mucosa during acute and chronic stress in the rat", In Vivo, 2(2), pp.143~146 (1988)
- Ryberg B, "Gastrin stimulates the self replication rate of the enterochromaffin-like cells in the rat stomach", Gastroenterology, 99, pp.935~942 (1990)
- Rytkonen J, "BCG vaccine modulates intestinal and systemic response to beta-lactoglobulin", Pediatr Allergy Immunol, 15(5), pp.408~414 (2004)
- Stevens CE, "Renewal of the mucous cell in the gastric mucosa of the rat", Anat Rec, 115, pp.231~243 (1953)
- Thompson FM, "Epithelial growth of the small intestine in human infants", J Pediatr Gastroenterol Nutr, 26(5), pp.506~512 (1998)
- Tielemans Y, "Proliferation of enterochromaffin like cells in omeprazole-treated hypergastrinemic rats", Gastroenterology, 96, pp.723~729 (1989)
- Varedi M, "A thermal injury-induced circulating factor(s) compromises intestinal cell morphology, proliferation, and migration", Am J Physiol, 277(1), pp.175~182 (1999)
- Yang X, "Systemic mycobacterial infection inhibits antigen-specific immunoglobulin E production, bronchial mucus production and eosinophilic inflammation induced by allergen", Immunology, 98(3), pp.329~337 (1999)
- Zembala M, "Tumour-cell-induced production of tumour necrosis factor by monocytes of gastric cancer patient receiving BCG immunotherapy", Cancer Immuol Immunother, 36, pp.127~132 (1993)