본문 바로가기
KERD
논문KCI 미등재

BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향

Effects of BCG on the DNA Synthesis and Ultrastructure of Mouse Gastric Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells

박경호|박대균|고정식|류인상
한국현미경학회 (Korean Society Of Microscopy)|2009.09|Applied Microscopy|제39권 제3호|pp. 205-218

1

조회

0

다운로드

1

인용

0

참고문헌

주제분류자연과학일반

초록

이 실험은 Ehrlich 종양세포를 이식한 후 BCG를 투여하였을 때, 위점막 상피세포의 형태학적 변화와 DNA합성능의 변화를 연구하 고자 시행하였다. 실험동물로는 체중 25 g 내외의 성숙한 생쥐(ICR계통)를 정상대 조군, 종양세포이식대조군(종양대조군), 종양세포이식후 BCG투여 군(BCG투여군)으로 구분하였다. 종양대조군과 BCG투여군 동물들 은 샅부위 피하에 각각 1×107의 Ehrlich 종양세포를 이식한 후, 다 음날부터 BCG (0.6×108~6.4×108 CFU, 27 mg/vial, Connaught Lab., Canada)를 하루건너 한 번씩 피부밑조직에 주사하였으며, 종 양대조군은 종양세포이식 후에 BCG 대신 0.2mL의 생리식염수를 피부밑조직에 주사하였다. 자기방사법적 관찰을 위해서는 BCG를 마지막으로 주사한 다음날 3H-thymidine (methyl-3H-thymidine: specific activity 25 Ci/mmol, Amersham Lab., England) 0.7μCi/gm를 꼬리정맥에 주사하고, 70분 후 도살하여 위조직을 떼어내어 10% formalin에 고정하였다. 자기방사표본관찰은 위점막 조직이 세로로 잘 절단된 부위를 택하여 점막근육판을 따라 점막길이 3.5mm의 위점막 조직에 분포하는 3H-thymidine 표지세포의 수를 계수하였으며, 일반조직 관찰을 위해서는 hematoxylin-eosin (H-E)염색을 시 행하였다. 전자현미경 관찰을 위해서는 떼어낸 위조직을 2.5% glutaraldehyde- 1.5% paraformaldehyde 혼합액에 고정한 후, 1% osmium tetroxide 액에 다시 고정하였으며, 고정이 끝난 조직은 탈수과정을 거쳐 araldite 혼합액에 포매하였다. 광학현미경적 관찰에서 종양대조군과 BCG투여군은 위점막 조 직에서 형태적으로 큰 변화를 볼 수 없었다. 전자현미경적 관찰에 서 BCG투여군의 점액상피세포는 전체적인 모습이 정상대조군과 종양대조군의 소견과 유사하였으나 정상대조군과 종양대조군에 비 해 수초구조와 뭇소포체(multivesicular body) 및 전자밀도가 높은 큰 미토콘드리아속과립이 자주 관찰되었다. 자기방사법적 관찰에 서 정상대조군, 종양대조군, BCG투여군은 점막길이 3.5 mm 당 출 현하는 표지세포수가 각각 380.2 (±31.35), 426.1 (±28.43) 및 301.8 (±34.63)개이었으며, BCG투여군은 정상대조군과 종양대조 군에 비하여 표지된 은입자의 수가 매우 적어서 표지과립이 겨우 구별될 정도의 세포가 많이 관찰되었다. 이상의 결과를 종합해보면 Ehrlich 종양세포를 이식한 동물에 BCG를 반복 투여하면 위점막 상피세포의 DNA합성을 효과적으 로 억제하면서도 형태적인 변화가 매우 경미하였으며, 이러한 결과 는 BCG가 항암치료 시 보조제로 사용할 수 있는 좋은 약제라고 생각된다.

Abstract

This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). 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 gastric 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. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in 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 380.2 (±31.35), 426.1 (±28.43) and 301.8 (±34.63), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.

참고문헌 (39)

  1. 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)
  2. Bartlett GL, "Suppression of murine tumor growth by immune reaction to the Bacillus Calmette-Guern strain of Mycobactrium bovis", J Natl Cancer Inst, 48, pp.245~257 (1972)
  3. Cameron IL, "Cell proliferation and renewal in the mammalian body in : Cellular and Molecular Renewal in the Mammalian Body", pp.45~85 (1971), Academic Press
  4. Chirigos MA, "Immunomodulators: Current and future development and application", Thymus, 19(1), pp.S7~S20 (1992)
  5. Chung JW, "Human Tissue Biology", pp.510~523 (2006), Soonoonsa Publishing
  6. DeHaven JI, "Antibiotic and steroid therapy of massive systemic Bacillus Calmette-Guerin toxicity", J Urol, 147, pp.738~742 (1992)
  7. El-Demiry MIM, "Local immune responses after intravesical BCG treatment for carcinoma in situ", Brit J Urol, 60, pp.543~548 (1987)
  8. Fawcett DW, "A textbook of histology", pp.460~472 (1994), Chapman and Hall
  9. Ghadially FN, "Ultrastuctural pathology of the cell and matrix", pp.310~313 (1997), Butterworths
  10. Hagen SJ, "Role of vacuolation in the death of gastric epithelial cells", Am J Physiol, 272(1), pp.C48~58 (1997)
  11. Helander HF, "The cells of the gastric mucosa", Int Rev Cytol, 70, pp.217~289 (1981)
  12. Kantani-Matsumoto A, "Mucus release of surface mucous cells of the mouse stomach with special reference to cell maturation stages and dietary conditions", Arch Histol Jpn, 50(3), pp.273~282 (1987)
  13. Karam SM, "Dynamics of epithelial cells in the corpus of the mouse stomach. II. Outward migration of pit cells", Anat Rec, 236(2), pp.280~296 (1993)
  14. Karam SM, "Origin and migratory pathways of the eleven epithelial cell types present in the body of the mouse stomach", Microsc Res Tech, 31(3), pp.193~214 (1995)
  15. Kataoka K, "Panoramic observation of the mouse gastric mucosa by superwide-field electron microscopy", Arch Histol Jpn, 47(2), pp.209~221 (1984)
  16. Kataoka K, "Histogenesis of the mouse gastric mucosa, with special reference to type and distribution of proliferative cells", Arch Histol Jpn, 47(5), pp.459~474 (1984)
  17. Kim MS, "Ultrastuructural alteration in the gastric chief cells of mouse, induced by 5-fluorouracil or mitomycin C", The Korean J Anat, 38(5), pp.421~431 (2005)
  18. Klimpel GK, "BCG-induced suppressor cells I. Demonstration of a macrophage-like suppressor cell that inhibits cytotoxic T cell generation in vitro", J Immunol, 120, pp.563~569 (1978)
  19. Ko JS, "Ahn ET: Effects of BCG or AG60 on gastric parietal cells of the mouse implanted with Ehrlich carcinoma cells", The Korean J Anat, 35(6), pp.8~542 (2002), 대한해부학회
  20. Ko JS, "Ultrastuructural alteration induced 5-fluorouracil or mitomycin C on the gastric parietal cells of mouse", The Korean J Anat, 35(5), pp.363~375 (2002)
  21. Kudo S, "The role of lymphocytes and histiocytes as mechanism of action of BCG of bladder cancer", Nippon Hinyokika Gakkai Zasshi, 84, pp.303~312 (1993)
  22. Lamm DL, "BCG in perspective: Advances in the treatment of superficial bladder cancer", Eur Urol, 27(2), pp.2~8 (1995)
  23. Lamm DL, "Enhanced natural killer (NK) cell activity with BCG and vitamin treatment (abstract 991)", J Urol, 151, pp.475A~ (1994)
  24. Lee ER, "Dynamic histology of the antral epithelium in the mouse stomach; IV. Ultrastructure and renewal of gland cells", Am J Anat, 172, pp.241~259 (1985)
  25. Magami Y, "Cell kinetics of slow renewing cell populations in mice stomach", J Gastroenterol Hepatol, 17, pp.262~269 (2002)
  26. Mathe G, "Immune manipulation by BCG administered before or after cyclophosphamide for chemoimmunotherapy of L1210 leukemia", Eur J Cancer, 10, pp.661~670 (1974)
  27. McKhann CF, "Immunotherapy of melanoma with BCG: Two fatalities following intralesional injection", Cancer, 35, pp.514~520 (1975)
  28. Miyauchi M, "Ontogeny of the rat parietal cell; Analysis using anti-parietal cell antibody and transmission electron microscopy", Kaibogaku Zasshi, 74(2), pp.197~207 (1999)
  29. Murayama Y, "Morphological and functional restoration of parietal cells in helicobactor pylori associated enlarged fold gastritis after eradicatin", Gut, 47(2), pp.313~314 (2000)
  30. Ogata T, "Morphological studies on the translocation of tubulovesicular system toward the intracellular canaliculus during stimulation of the gastric parietal cell", Microsc Res Tech, 48(5), pp.282~292 (2000)
  31. Ozogul C, "An ultrastructural investigation in stomach epithelial cells of mice during pregnancy and early lactation", Acta Gastroenterol Belg, 66(2), pp.137~143 (2003)
  32. Popiela T, "Postoperative immunochemotherapy (BCG+5-FU) in advanced gastric cancer", Anticancer Res, 8, pp.1423~1428 (1988)
  33. Ratliff TL, "Requirement of a thymus dependent immune response for BCG-mediated antitumor activity", J Urol, 137, pp.155~158 (1987)
  34. Roitt I, "Immunology", pp.33~44 (1993), Mosby-Year Book
  35. 류인상, "BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의미세구조에 미치는 영향", 한국현미경학회지, 35(3), pp.153~163 (2005), 한국현미경학회
  36. Rytkonen J, "BCG vaccine modulates intestinal and systemic response to beta-lactoglobulin", Pediatr Allergy Immunol, 15(5), pp.408~414 (2004)
  37. Suzuki H, "Ammonia-induced apoptosis is accelerated at higher pH in gastric surface mucous cells", Am J Physiol Gastrointest Liver Physiol, 283(4), pp.G986~995 (2002)
  38. 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)
  39. 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)

관련 논문AI추천

Bacillus Calmette-Guerin (BCG) 투여가 Ehrlich 종양세포를 이식한 생쥐 샘창자 상피세포의 DNA합성에 미치는 영향고정식, 김덕수, 김흥노, 박경호, 박대균한국현미경학회 · Applied Microscopy2010 · 제40권 제4호BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의미세구조에 미치는 영향고정식, 김명수, 류인상, 박경호, 박대균, 안의태한국현미경학회 · Applied Microscopy2005 · 제35권 제3호5-Fluorouracil, Mitomycin C 및 Acriflavine-Guanosine 복합제가 Ehrlich 암세포를 이식한 생쥐 위점막 점액상피세포의 미세구조에 미치는 영향고은주, 고정식, 김덕수, 박경호, 박대균한국현미경학회 · Applied Microscopy2011 · 제41권 제1호Adriamycin 및 AG60이 Ehrlich 종양세포를 이식한 생쥐 샘창자점막 상피세포의 DNA합성에 미치는 영향고정식, 김진국, 박경호, 안병준, 안의태대한체질인류학회 · 해부·생물인류학2002 · 제15권 제4호5-fluorouracil, mitomycin C, AG60이 Ehrlich 암세포를이식한 생쥐 곧창자점막 상피세포의 DNA 합성에미치는 영향고정식, 박경호, 박대균, 안의태, 정은주대한체질인류학회 · 해부·생물인류학2005 · 제18권 제3호종양치료제가 생쥐 막창자점막의 DNA 합성에 미치는 영향고정식, 나지웅, 박경호, 박대균, 안의태대한체질인류학회 · 해부·생물인류학2006 · 제19권 제4호

이 간행물의 다른 논문 (8건)

경골어류 시클리드과 Cichlasoma managuensis의 정자형성과정김석, 주경복, 이규재, 등영건, 장병수, 김동희, 송미숙pp. 219-223경골어류 잉어과 쉬리(Coreoleuciscus splendidus)의 정자형성과정등영건, 김석, 이규재, 김동희pp. 227-236경골어류 황어아과 버들치의 난자형성과정김동희, 이규재, 장병수, 김석, 정한석, 등영건pp. 237-243돌마자(Microphysogobio yaluensis)의 정소 내 생식세포에 관한 연구김재구, 김동희, 류동석pp. 245-252당뇨가 유발된 햄스터 창상치유에 미치는 벼세포 유래 GM-CSF의 효과허시현, 한규범, 정진주, 한만덕, 신길상, 김완종pp. 253-260
원문 보기