An aberrant spliced transcript of focal adhesion kinase is exclusively expressed in human breast cancer.
An aberrant spliced transcript of focal adhesion kinase is exclusively expressed in human breast cancer.
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粘着斑激酶的异常剪接转录物仅在人类乳腺癌中表达
DOI：
10.1186/1479-5876-12-136
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发表时间：
2014-05-21
影响因子：
7.4
通讯作者：
Shao Z
中科院分区：
文献类型：
--
作者：
Yao L;Li K;Peng W;Lin Q;Li S;Hu X;Zheng X;Shao Z
目的探讨一种新的FAK基因外显子26缺失的异常剪接转录本FAK在乳腺癌中的作用。方法采用逆转录聚合酶链式反应、DNA测序和琼脂糖凝胶电泳法对102例乳腺癌组织和52例正常乳腺组织中FAK基因的转录产物进行分析。克隆-26外显子FAK基因并在MCF-10A细胞中表达，分别用蛋白印迹法、免疫荧光染色和迁移实验检测FAK的活性、细胞定位和迁移能力。用Western blotting分析FAK在肿瘤坏死因子-α和去血清培养的MCF-10A细胞中的表达水平。结果-26外显子FAK转录本仅存在于人乳腺肿瘤组织中，其编码蛋白具有与野生型FAK相同的激酶活性、细胞定位和促进细胞迁移的能力。在用肿瘤坏死因子-α处理的MCF-7细胞和去血清的MCF-10A细胞中，-26外显子FAK对蛋白水解酶有抵抗作用，而野生型FAK大部分被切割。结论-26-外显子FAK转录本仅在人乳腺肿瘤组织中表达，其编码的蛋白具有与野生型FAK相同的激酶活性和生物学功能，但由于它对诱导野生型FAK蛋白分解的caspase介导的切割具有抵抗力，因此最终阻止了细胞的凋亡。
PurposeTo clarify the roles of a new aberrantly spliced transcript of FAK that lacks exon 26 (denoted -26-exon FAK) in human breast cancers.MethodsTranscripts ofFAKexpressed in 102 human breast tumor tissues and 52 corresponding normal tissues were analyzed by RT-PCR and DNA sequencing, as well as agarose gel electrophoresis. The cDNA of -26-exon FAK was cloned and expressed in MCF-10A cells, and then the kinase activity, cellular localization and migration capability of FAK were examined by western blotting, immunofluorescent staining and migration assays, respectively. The expression levels ofFAKwere analyzed by western blotting in MCF-7 cells treated with TNF-α or in MCF-10A cells upon serum deprivation. The MCF-10A cells transfected with a plasmid expressing -26-exon FAK were cultured in serum-free medium and cell apoptosis was analyzed by flow cytometry.ResultsThe -26-exon FAK transcript was exclusively present in human breast tumor tissues and the encoded protein possessed the same kinase activity, cellular localization and cell migration-promoting ability as wild-type FAK. In MCF-7 cells treated with TNF-α, and in MCF-10A cells upon serum deprivation, the -26-exon FAK was resistant to proteolysis while wild-type FAK was largely cleaved. In addition, the -26-exon FAK, but not wild-type FAK, inhibited cell apoptosis.ConclusionsThe -26-exon FAK transcript, which is exclusively expressed in human breast tumor tissues, encodes a protein that possesses the same kinase activity and biological function as the wild-type FAK, but because it is resistant to the caspase-mediated cleavage that induces the proteolysis of the wild-type form, it ultimately prevents apoptosis.
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DOI：
10.1084/jem.187.4.579
发表时间：
1998-02-16
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Levkau B;Herren B;Koyama H;Ross R;Raines EW
通讯作者：
Raines EW
影响因子：
5.3
作者：
SCHALLER, MD;BORGMAN, CA;PARSONS, JT
通讯作者：
PARSONS, JT
影响因子：
5.5
作者：
Zhang, HFM;Keledjian, KM;Wang, JY
通讯作者：
Wang, JY
影响因子：
4.4
作者：
Corsi JM;Rouer E;Girault JA;Enslen H
通讯作者：
Enslen H
影响因子：
4.8
作者：
Sonoda, Y;Watanabe, S;Kasahara, T
通讯作者：
Kasahara, T