| 摘要: |
| 目的:建立可稳定传代的肾透明细胞癌小鼠肺转移模型，并在此基础上初步筛选出肾透明细胞癌特异性鼠肺转移细胞亚株，为后续研究奠定基础。方法:选用本实验室前期建立的肾透明细胞癌细胞株RCC05-TXJ，体外扩增培养制成细胞悬液后注射于NOD-SCID小鼠颈背部皮下，成瘤后取出瘤组织接种于另外NOD-SCID小鼠的肾包膜下，待有肉眼可见瘤块后，处死小鼠并取出肿瘤组织接种于新NOD-SCID小鼠的肾包膜下，反复数轮，直至出现稳定的鼠肺转移模型。此时取模型鼠肺转移组织，一部分继续接种于新鼠肾包膜下，反复数轮，尝试以转移组织诱导肺转移；另一部分进行原代培养，细胞经体外扩增纯化后制成悬液接种于新鼠肾包膜，数轮循环至产生肺转移，筛选能产生稳定鼠肺转移的细胞亚株。整个筛选过程均观察记录原位成瘤、淋巴及肺转移形成情况，并对肿瘤组织进行病理组织学观察及CA9（肾癌标志）、CD133（干细胞标志）表达检测。结果:以RCC05-TXJ细胞株接种所致NOD-SCID小鼠颈背部皮下肿瘤组织作为移植瘤材接种鼠肾包膜在第2轮即出现鼠肺转移，此后3~6轮以鼠肺转移组织移植肾包膜下，均能成功诱导转移。以鼠肺转移组织原代培养RCC细胞均获成功，第6轮原代培养细胞扩增纯化后制成的细胞悬液（RCC05-TXJ-L）接种于鼠肾包膜下，出现特异性肺转移，重复2轮实验结果稳定。RCC05-TXJ-L细胞具有生长速度快（传代时间约为2 d）、原位成瘤时间短（1周左右）、接种后转移性较强、CA9及CD133表达升高等特点。结论:RCC05-TXJ在NOD-SCID小鼠体内多轮反复筛选可建立稳定的肾细胞癌鼠肺转移动物模型；在此基础上筛选出的肺转移性细胞亚株RCC05-TXJ-L可稳定诱导NOD-SCID鼠产生肺部转移。 |
| 关键词: 肾细胞癌；肺转移；动物模型 肺转移细胞亚株 |
| DOI：10.3724/SP.J.1008.2008.01446 |
| 投稿时间：2008-03-07修订日期：2008-05-17 |
| 基金项目:国家自然科学基金（30571609,30873041）. |
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| Establishing a lung metastasis mouse model of clear cell renal cell carcinoma and screening of lung metastasis sub-cell line |
| ZHAI Yu-jia1,TAN Xiao-jie1,LIN Li-ping1,HOU Jian-guo2,WU Qi2,CHANG Wen-jun1,MA Li-ye3,YAO Ming4,CAO Guang-wen1*,CAO Guangwen |
| (1.Department of Epidemiology,Faculty of Health Service,Second Military Medical University,Shanghai 200433,China* 2.Department of Urology,Changhai Hospital,Second Military Medical University,Shanghai 200433*3.Department of General Surgery,Changhai Hospital,Second Military Medical University,Shanghai 200433* 4.Center of Experimental Animals,Shanghai Cancer Institute,Shanghai 200003) |
| Abstract: |
| Objective:To establish a lung metastasis mouse model of clear cell renal cell carcinoma (CCRCC) and screen for lung metastasis CCRCC cell line,so as to lay a foundation for future study.Methods: RCC05-TXJ,a CCRCC cell line established by our lab,was used in the present study.The cells were proliferated and the cell suspension was subcutaneously injected into the NOD-SCID mice.The newly-formed tumors were transplanted into the renal capsule of NOD-SCID mice (in situ),and the formed tumors were harvested and again transplanted into NOD-SCID mice.A stable mouse model with CCRCC lung metastasis was established following several cycles of the transplantation.This transplantation cycle was repeated until the development of stable lung metastasis.The lung metastasis tissues were then injected into renal capsule for several cycles to induce the lung metastasis.The metastasis tissues were also primary cultured and proliferated and the cell suspensions were injected into renal capsule until the development of lung metastasis.The sub-cell lines which could stably produce lung metastasis were screened.During the screening,the in situ tumor information,lymphatic and pulmonary metastasis were all recorded in detail.Immunohistochemistry analyses were used to examine the expression of CA9 (a marker of renal cells) and CD133 (a marker of stem cells) on the metastatic CCRCC cells.Results: The lung metastasis was observed in the second cycle of the in situ transplantation of RCC05-TXJ in NOD-SCID mice.When the metastatic lung carcinoma tissues were used for in situ transplantation,lung metastasis was successfully established during the 3-6 cycles.CCRCC cells were successfully cultured with lung metastatic tissues,and the cell suspension of the passage 6 cells caused lung metastasis after implanted in the renal capsule.The result was stable in two repeated cycles.The lung metastatic RCC05-TXJ-L cells grew fast (average passage time was 2 days),and had a short in situ tumor-forming time (1 week).They also had a strong potential for lung metastasis and high expression of CA9 and CD133.Conclusion: Repeated in situ transplantation of RCC05-TXJ can be used to establish stable lung metastasis mice model of CCRCC.The selected RCC05-TXJ-L sub-cell line can stably induce lung metastasis in NOD-SCID mice. |
| Key words: renal cell carcinoma lung metastasis animal model lung metastasis sub-cell line |
