摘要
背景与目的：乳腺癌细胞系大部分来源于乳腺癌转移所致的胸水或非原始乳腺癌组织来源，与真正的乳腺癌的生物学性状有很大差异。而仅部分细胞系直接由乳腺癌原代细胞在体外培养条件下成为永生化的细胞系，过表达人端粒酶逆转录酶（hTERT）可使正常人上皮细胞永生化。本研究将通过hTERT转染人原始乳腺癌组织来源的细胞建立永生化乳腺癌细胞系，并研究过表达hTERT对其他基因表达的影响。方法：通过重组逆转录病毒pBABE-hTERT-puro将hTERT基因导人原始乳腺癌组织来源的细胞，建立稳定表达hTERT的永生乳腺癌细胞。再利用微阵列技术检测乳腺癌细胞中hTERT诱导的基因表达改变。结果：Real-TimeRT-PCR证实hTERT转染后的乳腺癌细胞中hTERT过度并稳定表达，为转染前表达量的1400倍以上，并使得被转染的细胞永生化，可以传40～50代以上。微阵列结果进一步证实hTERT在转染后的细胞中过表达，另外，有22个基因在转染后的乳腺癌细胞中上调，未观察到所有细胞株中均下调的基因。结论：过表达hTERT可导致乳腺癌原代细胞的永生化，并对部分基因有上调作用。
Background and purpose： Most breast cancer cell lines were derived from malignant pleural fluid or other tissues that metastasized from breast carcinoma. Breast carcinogenesis is under represented due to the significant distinction of biological characteristics between cancer cell lines and primary breast cancers. Few primary breast tumors spontaneously developed into immortal cell lines. Overexpression of human telomerase reverse transcriptase （hTERT） in normal epithelial cells led to immortalization. We aimed to establish immortalized cultures derived from primary breast cancer by introducing hTERT and determine the gene profile of hTERT-transfected cultures. Methods： hTERT was introduced into breast tumor cultures with a puromycin-resistant retroviral construct, pBABE-hTERT-puro. Real-Time RT-PCR was used to analyze the expression level of hTERT after transfection. The up-regulated gene profile was determined with microarray techniques. Results： hTERT was stably over expressed, at least 1 400-fold in hTERT-transfected cells compared to pBABE-puro transfected cultures. These ectopic over-expressions led to immortalization of transfected cultures, which could undergo at least 40-50 passages. Microarray analysis further confirmed the over-expression of hTERT in transfected cultures and demonstrated that 22 genes were up-regulated due to over-expression of hTERT. No down-regulated genes were observed cross all cultures. Conclusions： Over-expression of hTERT led to immortalization of breast tumor primary cultures and up-regulated certain genes.
出处
《中国癌症杂志》
CAS
CSCD
2007年第2期125-129,共5页
China Oncology
关键词
端粒酶逆转录酶
原始乳腺癌
细胞培养
永生化
基因表达谱
human telomerase reverse transcriptase
primary breast tumor
cell cultures
immortalization
gene profile
作者简介
通讯作者：白春学E-mail：cxbai@zshospital．com
