南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (05): 664-.

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肺癌骨转移小鼠模型的建立

孟越,李春雨,郝松,胡少宇,林振,袁亮,李威,闫文娟,陈建庭,杨德鸿   

  • 出版日期:2014-05-20 发布日期:2014-05-20

Bone metastasis of lung cancer in a mouse model with normal immune function

  • Online:2014-05-20 Published:2014-05-20

摘要: 目的通过短暂免疫抑制,在免疫功能正常小鼠复制肺癌骨转移模型。方法6周龄雌性C57BL/6J小鼠40只,随机分为对
照组(CON)、低剂量免疫抑制组(C100/D50)、中等剂量免疫抑制组(C150/D100)及高剂量免疫抑制组(C200/D150)。低剂量免
疫抑制组、中等剂量免疫抑制组及高剂量免疫抑制组在实验的第1天分别腹腔注射环磷酰胺(CTX)100、150、200 mg/kg,在次
日分别注射地塞米松钠(DXM)50、100、150 mg/kg,连续注射3 d。对照组小鼠腹腔注射等量生理盐水。第5天,取对数生长期
的人肺腺癌A549 细胞,注射1.0×109个/L浓度的细胞悬液于小鼠右后肢胫骨平台下。造模后第28天进行影像学测定并取材,
评估骨质缺损程度,HE染色及免疫组织化学法观察肿瘤形态,并通过检验学指标评估骨破坏程度。结果在肿瘤注射过程中除
对照组外每组各有1只死亡。试验结束时对照组小鼠全部存活;C100/D50组小鼠9只存活,仅有1只成瘤;C150/D100组小鼠存
活8只,7只成瘤;C200/D150组小鼠存活4只,全部成瘤。中、高剂量免疫抑制组的小鼠体重降低明显,与其他组比较,差异有显
著性意义(P<0.05)。X线及显微CT可见中、高剂量免疫抑制组小鼠胫骨平台下出现明显的骨缺损与骨破坏,其影像学表现符
合肿瘤性骨破坏的改变。病理可见溶骨病变为主的混合性骨质破坏。中、高剂量免疫抑制组小鼠ALP活性明显高于其他两组,
差异有显著性意义(P<0.05),但这两组间之间无明显差异。结论中、高剂量免疫抑制剂在制备小鼠肺癌骨转移模型中均具有
较高的成瘤率,但中等剂量组小鼠存活率高,效果更理想。建立了比较成熟的人肺腺癌细胞移植瘤模型制备方法,为转移瘤的
研究和治疗提供动物模型。

Abstract: Objective To establish a model bearing human lung cancer xenograft with bone metastasis in mice with normal
immune function. Methods Forty female C57BL/6J mice were randomly allocated into 4 equal groups, including a control
group and 3 immunosuppression groups treated with low, moderate, and high doses of dexamethasone (50, 100, and 150 mg,
respectively). Four days after immune suppression, the mice were subjected to percutaneous injection of1.0×109 L-1 A549 cells
into the tibial plateau, and the bone defects were assessed radiographically 28 days after modeling. HE staining and
immunohistochemical staining were used to examine the tumor tissues and bone tissue damages. Results In each of the 4
groups one mouse died during tumor cell injection. Only 1 mouse showed tumor formation in low-dose immunosuppression
group, as compared to 7 and 4 in moderate- and high-dose immunosuppression groups. X-ray and microCT scan showed
significant tibial bone destruction in moderate- and high-dose groups. The moderate- and high-dose groups showed similar
ALP activities but both were significantly higher than those in the other two groups (P<0.05). Conclusion Immunosuppression
with a moderate dose of dexamethasone results in longer survival time of the human lung cancer xenograft-bearing model
mice as well as a higher tumor formation rate.