中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2011年
4期
246-250
,共5页
于宝华%周晓燕%张铁成%张太明%施达仁
于寶華%週曉燕%張鐵成%張太明%施達仁
우보화%주효연%장철성%장태명%시체인
淋巴瘤,大细胞,弥漫型%肿瘤移植%细胞,培养的%模型,动物
淋巴瘤,大細胞,瀰漫型%腫瘤移植%細胞,培養的%模型,動物
림파류,대세포,미만형%종류이식%세포,배양적%모형,동물
Lympthoma,large-cell,diffuse%Neoplasm transplatation%Cell,cultured%Model,animal
目的 建立人弥漫性大B细胞淋巴瘤(DLBCL)细胞株LY8裸鼠皮下移植瘤模型并观察其生长特性,为探讨淋巴瘤发病机制及治疗策略提供手段.方法 将人DLBCL细胞株LY8种植于裸鼠右前肢肩胛背侧皮下,成瘤后无菌套管针抽吸法抽取约1.5 mm×1.5 mm×1.5 mm大小组织块进行皮下种植,观察成瘤率和组织形态特点;用免疫组织化学EnVision法观察白细胞共同抗原(LCA)、CD20、CD79α、Ki-67、CD3、CD45RO、bcl-6、MUM-1、CD10、bcl-2等指标的表达;用聚合酶链反应(PCR)扩增检测移植瘤和LY8细胞株的IgH基因克隆性重排和针对3个微卫星位点(D14S68、D18S69、D20S199)的基因组DNA微卫星序列.结果 运用LY8细胞株成功进行了人DLBCL的种植,且生长稳定,已传至第9代,共移植裸鼠124只,其中114只成瘤,成瘤率达91.9%.每代移植瘤的生长特点均相似,于移植后约2周长出,约在3周左右长至最大径1.3 cm,随后即进入快速生长期,4周左右达2.0 cm大小.移植瘤形态符合DLBCL,肿瘤细胞免疫组织化学显色呈LCA、CD20、CD79α、bcl-6、MUM-1、CD10、bcl-2阳性;裸鼠移植瘤与LY8有相同的IgH基因克隆性重排,经3个位点的微卫星引物扩增后获得相同片段的产物.表明移植瘤与人DLBCL细胞相似,并证实了其生长的稳定性和可重复性.结论 建立了可稳定传代的DLBCL的裸鼠模型,为研究人DLBCL的生物学特性及试验治疗提供了较理想的动物模型.
目的 建立人瀰漫性大B細胞淋巴瘤(DLBCL)細胞株LY8裸鼠皮下移植瘤模型併觀察其生長特性,為探討淋巴瘤髮病機製及治療策略提供手段.方法 將人DLBCL細胞株LY8種植于裸鼠右前肢肩胛揹側皮下,成瘤後無菌套管針抽吸法抽取約1.5 mm×1.5 mm×1.5 mm大小組織塊進行皮下種植,觀察成瘤率和組織形態特點;用免疫組織化學EnVision法觀察白細胞共同抗原(LCA)、CD20、CD79α、Ki-67、CD3、CD45RO、bcl-6、MUM-1、CD10、bcl-2等指標的錶達;用聚閤酶鏈反應(PCR)擴增檢測移植瘤和LY8細胞株的IgH基因剋隆性重排和針對3箇微衛星位點(D14S68、D18S69、D20S199)的基因組DNA微衛星序列.結果 運用LY8細胞株成功進行瞭人DLBCL的種植,且生長穩定,已傳至第9代,共移植裸鼠124隻,其中114隻成瘤,成瘤率達91.9%.每代移植瘤的生長特點均相似,于移植後約2週長齣,約在3週左右長至最大徑1.3 cm,隨後即進入快速生長期,4週左右達2.0 cm大小.移植瘤形態符閤DLBCL,腫瘤細胞免疫組織化學顯色呈LCA、CD20、CD79α、bcl-6、MUM-1、CD10、bcl-2暘性;裸鼠移植瘤與LY8有相同的IgH基因剋隆性重排,經3箇位點的微衛星引物擴增後穫得相同片段的產物.錶明移植瘤與人DLBCL細胞相似,併證實瞭其生長的穩定性和可重複性.結論 建立瞭可穩定傳代的DLBCL的裸鼠模型,為研究人DLBCL的生物學特性及試驗治療提供瞭較理想的動物模型.
목적 건립인미만성대B세포림파류(DLBCL)세포주LY8라서피하이식류모형병관찰기생장특성,위탐토림파류발병궤제급치료책략제공수단.방법 장인DLBCL세포주LY8충식우라서우전지견갑배측피하,성류후무균투관침추흡법추취약1.5 mm×1.5 mm×1.5 mm대소조직괴진행피하충식,관찰성류솔화조직형태특점;용면역조직화학EnVision법관찰백세포공동항원(LCA)、CD20、CD79α、Ki-67、CD3、CD45RO、bcl-6、MUM-1、CD10、bcl-2등지표적표체;용취합매련반응(PCR)확증검측이식류화LY8세포주적IgH기인극륭성중배화침대3개미위성위점(D14S68、D18S69、D20S199)적기인조DNA미위성서렬.결과 운용LY8세포주성공진행료인DLBCL적충식,차생장은정,이전지제9대,공이식라서124지,기중114지성류,성류솔체91.9%.매대이식류적생장특점균상사,우이식후약2주장출,약재3주좌우장지최대경1.3 cm,수후즉진입쾌속생장기,4주좌우체2.0 cm대소.이식류형태부합DLBCL,종류세포면역조직화학현색정LCA、CD20、CD79α、bcl-6、MUM-1、CD10、bcl-2양성;라서이식류여LY8유상동적IgH기인극륭성중배,경3개위점적미위성인물확증후획득상동편단적산물.표명이식류여인DLBCL세포상사,병증실료기생장적은정성화가중복성.결론 건립료가은정전대적DLBCL적라서모형,위연구인DLBCL적생물학특성급시험치료제공료교이상적동물모형.
Objective To establish a diffuse large B-cell lymphoma (DLBCL)-mice model using human DLBCL cell line LY8, to investigate its characteristics of growth and to provide a model for in vivo study of DLBCL pathogenesis and treatment. Methods LY8 cells were injected subcutaneously into the right flank of nude mice. Harvested tumor tissues were cut into small pieces of 1.5 mm × 1.5 mm × 1.5 mm and implanted subcutaneously into nude mice. Tumor growth was visualized and the histologic characteristics were documented. Expression of LCA, CD20, CD79α, Ki-67, CD3, CD45RO, bcl-6, MUM-1, CD1O and bcl-2 were examined by using immunohistochemistry. IgH clonal rearrangement and status of three microsatellite loci (D14S68, D18S69, D20S199) in the xenografted tumor samples and the parental cell line LY8 were detected using PCR amplification followed by PAGE. Results The subcutaneous xenograft DLBCL model was successfully established by using cell line LY8, and a stable growth was achieved up to the 9th generation. The tumor in each generation showed similar growth characteristics and the rate of subcutaneous tumor formation was 91.9% (114/124). The tumor growth was observed from the 2nd week after implantation, reaching 1.3 cm in major diameter at the 3rd week and 2. 0 cm at the 4th week. The tumor had identical morphological characteristics with those of human DLBCL, and expressed LCA, CE0,CD79α, bcl-6, MUM-1, CD1O and bcl-2. The tumor of xenograft mice and cell line LY8 showed identical IgH rearrangement and microsatellite length. Conclusions A human DLBCL bearing mouse model was successfully established. The mice model is similar to human counterpart with high stability and repeatability. Therefore, it provides an ideal animal model for in vivo studies of the biological characteristics and treatment of DLBCL.
