不同表型的人宫颈癌细胞亚克隆株的基因表达谱

目的　采用基因芯片技术分析同一人宫颈癌细胞株来源 ,锚着依赖性不同的两个亚克隆细胞株的侵袭相关基因表达。方法　采用有限稀释法建立CS12 13细胞的CS0 3和CS0 7亚克隆细胞株 ,检测其在软琼脂中的集落形成率 ,分别用流式细胞术和免疫组化法观察细胞黏附分子CD4 4和E cadherin、Fibronetin的表达 ,并将CS0 3和CS0 7细胞的总RNA经逆转录获得的cDNA作为探针 ,与含 10 11个与细胞信号转导和细胞膜受体相关基因表达谱芯片杂交 ,信号用ScanArray 30 0 0扫描分析。结果　CS0 3和CS0 7细胞的集落形成率分别是 0和 38.5 % ,CD4 4表达分别是 35 .1%和 13.7% ;两者的基因表达谱差异明显 ,CS0 3细胞中表达上调的基因为 12个 ,CS0 7细胞中有 2 1个基因表达增加 ,其中一些与细胞增生、迁移、凋亡 ,细胞间信号传导和肿瘤转移相关 ,包括p34cdc2 、转化生长因子 β刺激蛋白 (TSC2 2 )、纤维蛋白酶原活性因子抑制剂 (PAI 1)和桥粒相关蛋白 (PININ)等。结论　同一来源的人宫颈癌细胞株中有多个差异表达基因 ,它们参与人宫颈癌细胞的表型特征 ,可能影响肿瘤的进展。

Study on gene expression profile of subclonal cell lines in two different anchorage-independent uterine cervical carcinoma

Objective To examine the differently expressed in vasion-related genes in two anchorage-independent uterine cervical carcinoma c ell lines derived from the same patient using a cDNA array. Methods Two human uterine cervical carcinoma subclonal cell lines CS03 and CS07 derived from a single donor line CS1213 were established by limited diluting procedure and their cloning efficiency was counted in soft aga r. The expression of cellular adhesive molecules CD44, E-cadherin and Fibroneti n were studied by flow cytometry and immunohistochemical staining. The two cDNA samples retro-transcribed from total RNA derived from CS03 and CS07 cells were screened by a cDNA microarray carrying 1011 human signal transduction and membra ne receptor-associated genes and scanned with a ScanArray 3000 laser scanner. Results The cloning efficiency was 38.5% in CS07 cells and 0 in CS03 cells, respectively. CD44 were 35.1% and 13.7% in CS03 and CS07,respe ctively. The cDNA microarray analysis showed that 12 genes in CS03 were upregula ted compared with CS07, and 21 genes in CS07 were upregulated. The function of a number of differently expressed genes is consistently associated with cell prol iferation, migration, apoptosis, signal transduction and tumor metastasis, inclu ding p34 cdc2, TSC22, plasminogen activator inhibitor I (PAI-1)and desmoso me associated protein (Pinin). Conclusion Multiple genes are differently expressed in uter ine cervical carcinoma cell lines even in the same patient. It is suggested that these genes are involved with the phenotypic characteristics and development of cervical carcinoma.