肿瘤基质成纤维细胞模型的建立及生物学特性研究  

Establishment and identification of tumor fibroblast model

作　　者：朱红梅[1] 汤为学[2] 周卫平[1] 张晓艳[2] 马静[2] 郑维萍[2] 

机构地区：[1]重庆医科大学附属第二医院,重庆医科大学肝炎研究所 [2]重庆医科大学基础医学部病理生理学教研室,重庆400010

出　　处：《第三军医大学学报》2005年第1期31-35,共5页Journal of Third Military Medical University

摘　　要：目的　建立肿瘤基质成纤维细胞模型并探讨其恶性生物学特性 ,为研究其恶性转化机制、开发靶向肿瘤基质的抗癌药物提供有价值的细胞模型。方法　用小鼠肝癌细胞H2 2 培养上清液诱导小鼠成纤维细胞L92 9,获得能稳定生长的L92 9 H2 2 细胞。用光镜、电镜观察其形态改变 ,MTT法检测其生长情况 ,免疫细胞化学测定其PCNA、bcl 2、MMP 9、TN表达变化 ,RT PCR测定其端粒酶活性 ,并分析其核型、蛋白合成能力、对常用抗癌药物的反应性及其条件培养基 (conditionedmedium ,CM)对H2 2 细胞生长的影响。结果　L92 9 H2 2 细胞出现多核和畸形核 ,染色体众数略有增加。细胞群体倍增时间较亲代细胞缩短 (t=2 65 41,P <0 0 5 ) ,分裂指数增高 (χ2 =4 5 2 5 ,P <0 0 5 ) ,PCNA、bcl 2、MMP 9、TN表达增强 (t≥ 3 3 0 5 5 ,P<0 0 1)。端粒酶活性增高 (t=-4 0 4163 ,P <0 0 0 1)。L92 9 H2 2 细胞对作用于S期或S期和M期或细胞骨架的抗癌药物较L92 9敏感 (q≥ 3 0 15 2 ,P <0 0 5 )。蛋白质合成能力增强 (q =-5 90 11,P <0 0 0 1)。其CM促进H2 2 细胞生长。结论　L92 9 H2 2 细胞较亲代细胞增殖快 ,核型发生变化 ,存活能力强 ,功能活跃 ,表达并分泌促进肿瘤细胞生长与侵袭的因子增强 。Objectives To establish the cell model of fibroblast in tumor and explore its malignant bionomics for providing valuable cell model for studying their transforming mechanism and developing drugs targeting the tumor stroma in the future. Methods The murine normal fibroblast cells, L 929 , were cultured with the supernatant of murine hepatocellular carcinoma cells, H 22 , for three and a half months, and L 929 -H 22 cells were acquired with stable growth. Then the changes of their biological characteristics were determined by light microscopy, electron microscopy, karyotype analysis, and MTT assay. The expressions of PCNA, bcl-2, MMP-9, TN, and tolomerase activity were detected by immunocytochemistry and RT-PCR, respectively. The ability to synthesize proteins and the effects of their supernatant on H 22 growth were determined. Results Abnormal nuclei and multinuclei were observed in L 929 -H 22 cells, and their chromatosome modes increased slightly. As compared with the parental cells, the population doubling time of L 929 -H 22 cells shortened (t=2.654 1, P<0.05), mitotic indices increased (χ 2=4.525, P<0.05), the expressions of PCNA, bcl-2, MMP-9, and TN enhanced (t≥3.305 5, P<0.01). L 929 -H 22 cells were more sensitive than L 929 to anticancer drugs with inhibitory effect on S-phase or S- and M-phase or cell skeleton (q≥3.015 2, P<0.05). The ability to synthesize proteins and telomerase activity enhanced (q=-5.901 1, P<0.001; t= -40.416 3 , P<0.001). Their supernatant promoted H 22 cells growth. Conclusion The results suggest that L 929 -H 22 cells have some biological characteristics of the fibroblasts derived from tumors, including more quick proliferation, longer survival time and more functions, stronger expression and secretion of some growth factors, and can act as a cell model for developing drugs targeting tumor stroma in the future.

关 键 词：仲瘤 成纤维细胞 细胞模型 L929-H22 

分 类 号：R-331[医药卫生] R73-354