文档介绍：TGFβ1、TGFβR1、Smad4在乳腺癌发生、发展中的作用及其相互关系
(作者:___________单位: ___________邮编: ___________)
作者:夏立平,刘晓力,陈,何冬雷,高炳玉
【摘要】目的:探讨TGFβ1、TGFβR1及Smad4三者在乳腺癌中的表达与其发生、发展、转移等生物学行为以及预后的关系,为乳腺癌发病和治疗机制提供理论和实验依据。方法:提取30例乳腺癌临床标本(经液氮、研磨等处理)和MCF7、T47D、SKBR3三种乳腺癌细胞(取指数增长期细胞)总RNA,反转录cDNA第一链,克隆TGFβR1、Smad4基因片段,测定质粒浓度,制备标准样品,并作出标准曲线,探针法荧光定量PCR(FQPCR)检测TGFβR1和Smad4基因mRNA表达。结果:FQPCR检测发现恶性程度高的SKBR3细胞,其TGFβR1和Smad4的基因拷贝数明显低于恶性程度低的MCF7细胞(P均<),T47细胞两种基因拷贝数介于两者之间。结论:随着乳腺癌细胞病理类型恶性程度增加,TGFβR1和Smad4 mRNA表达逐步下降,表明TGFβ1和Smad4可能具有类似乳腺癌抑癌基因的作用。
【关键词】转化生长因子
β1; 转化生长因子β1受体; Smad4; 乳腺癌
[ABSTRACT] Objective: To investigate the expression of TGFβ1, TGFβR1 and Amad4 proteins and the relationship to the initiation, progression, metaptosis and prognosis of breast cancer. Methods: The samples of 30 cases and 3 cell lines(MCF7,T47D,SKBR3) of breast cancer were selected, and the total mRNA was extracted. Reverse transcription, and TGFβR1 and Smad4 genes cloning were carried out. Then the concentration of plasmid was measured, standard curve was made, and the expression of TGFβR1 and Smad4 mRNAs were assayed by FQPCR machine. Results: The gene copies of TGFβR1 and Smad4 in SKBR3 cells were lower than that in MCF7 cells (P<) and the gene copies in T47 was middle. Conclusions: Our studies by FQPCR show that the lower TGFβR1 and Smad4 mRNAs expression, the worse biology characteristic of breast cancer, which indicate TGFβR1 and Smad4 act possibly as the role of antioncogene in breast cancer.

[KEY WORDS] TGFβ1; TGFβR1; Smad4; Breast cancer
乳腺癌是女性常见恶性肿瘤之一,全世界每年约有120万妇女患乳腺癌。目前,乳腺癌病因尚不十分清楚,发病机理非常复杂,涉及多个因素的作用。多数研究认为,乳腺癌的发生是因为原癌基因的激活和抑癌基因的功能丧失,其发展是一个多因素多阶段的过程,往往涉及多个基因的改变[1,2]。TGF
β/Smads信号传导通路及其与肿瘤关系多见于胰腺癌、妇科肿瘤等[3,4],而在乳腺癌中TGFβ1、TGFβR1及Smad4三者的表达及其相互关系如何,它们对乳腺癌发生、发展有何意义,目前还不完全清楚。本实验拟采用荧光定量PCR (fluorescent quantitation polymerase chain reaction,FQPCR)、免疫组织化学/免疫细胞化学、Western Blotting等技术,检测乳腺癌患者和乳腺癌细胞株中的TGFβ1、TGFβR1和Smad4 在mRNA水平和蛋白水平表达,进一步深入三者在乳腺癌中的表达与其发生、发展、转移等生物学行为以