趋化因子IP10抑制乳腺癌细胞4T1播散转移的研究  

Inoculation of the cells transfected with IP10 inhibited experimental tumor metastases in vivo

作　　者：杨秀利[1] 储以微[1] 徐林[1] 李宝华[1] 熊思东[1] 

机构地区：[1]复旦大学上海医学院免疫学系教育部分子医学重点实验室,200032

出　　处：《中华医学杂志》2006年第3期176-181,共6页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30170867);国家教育部科学技术重点资助项目(03063)

摘　　要：目的观察增强IP10在肿瘤局部的表达对乳腺癌细胞4T1远端播散转移的作用。方法用电转染的方法建立稳定表达IP10的4T1细胞株IP10-4T1。将BALB/c小鼠分为IP10-4T1细胞组、4T1细胞组和转染pcDNA3的4T1细胞组(pcDNA3-4T1)。利用实验性肺转移模型和体外杀伤实验研究接种IP10-4T1对4T1细胞播散转移的影响。结果IP10-4T1细胞中IP10mRNA转录水平相对含量为0.92±0.12与未转染的4T1细胞(0.35±0.12)和pcDNA3-4T1细胞(0.29±0.08)比较差异有统计学意义(P<0.01)。IP10-4T1组对淋巴细胞的趋化指数为2.77±0.29,经CXCR3抗体处理后下降为1.71±0.20(P<0.05)。实验性肺转移结果显示,IP10-4T1组小鼠肺重为0.27g±0.02g,与4T1细胞组(0.48g±0.08g)和pcDNA3-4T1组(0.43g±0.16g)比较明显减轻(P=0.021);其肺表面形成的转移结节数较对照组少;IP10-4T1组肺组织中形成的4T1细胞克隆数为2.6±1.7,明显低于4T1组(34.0±6.3)和pcDNA3-4T1组(33.0±2.3,P<0.05);2/6的IP10-4T1组小鼠肺组织内可见转移灶,而对照组全部形成肺转移灶。接种IP10-4T1细胞组小鼠的淋巴细胞杀伤率在各个效/靶比均高于对照组(P<0.05)。结论增加肿瘤局部IP10的表达,能增强机体细胞免疫应答水平,通过肿瘤特异性的杀伤作用,抑制肿瘤细胞在体内播散转移。Objective To explore the inhibitory effects of IP10 on the experimental tumor metastases of a mammary carcinoma cell line 4T1 in vivo. Methods 4T1 cells were transfected with pcDNA3-IP10 plasmid and the positive clones （IP10-4T1） were screened in the presence of G418. The parental 4T1 cells and 4T1 cells transfected with pcDNA3 （pcDNA3-4T1） were used as controls. The expression of IP10 mRNA was examined with RT-PCR. The chemotactic activity of the cell-cultured supernatant for the activated lymphocytes was assessed with chemotaxis assay. All BALB/c mice were divided into 3 equal groups： IP10-4T1, parental 4T1 and pcDNA3-4T1 （6 mice in each group）. Seven days after mice were inoculated with 2 × 10^6 4T1, pcDNA3-4T1 or IP10-4T1 cells, 1 × 10^5 4T1 cells were injected into mice in tail vein. On day 14 tumors were sectioned. On day 28 mice were sacrificed and the lung weight, metastatic forci on the lung surface, disseminated metastases in the lungs and the number of clonogenic metastases of 4T1 cells were observed. The splenocytes were isolated from tumor-bearing mice, and the cytotoxicity of the splenocytes was evaluated by CFSE/7-AAD method. Results The transcription of IP10 mRNA increased in IP10-4T1 cells compared to parental 4T1 and pcDNA3-4T1 cells （P =0. 002）. Moreover, accumulated lymphocytes migrated to the supernatants of IP10-4T1 cells, which can be abrogated by anti-CXCR3 （P 〈 0.05）. Compared to controls, inoculation of IP10-4T1 cells resulted in the decreased disseminated metastases as indicated by dramatically reduced lung metastatic forci on the surface of the lungs; the lung weight was lighter （IP10-4T1,0.27 ±0.02 g v. s. 4T1,0.48 ±0.08 g and pcDNA3-4T1, 0.43 ±0. 16 g, P = 0. 021 ）; the number of clonogenic metastatic 4T1 cells enumerated in ten fields decreased greatly （ IP10-4T1,2.6 ± 1.7 v. s. 4T1, 34 ± 6.3 and pcDNA3-4T1, 33 ± 2.3, P 〈 0.05 ） ;histological assay showed that metastasis was not found in the lungs of the 4/6 of mice in IP10-4T1 group, and wa

关 键 词：乳腺肿瘤 肿瘤转移 细胞培养 

分 类 号：R737.9[医药卫生—肿瘤] R737.902[医药卫生—临床医学]