Suppression of breast cancer cell growth by Na+/H+ exchanger regulatory factor 1 (NHERF1).
Suppression of breast cancer cell growth by Na+/H+ exchanger regulatory factor 1 (NHERF1).
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DOI：
10.1186/bcr1616
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发表时间：
2006
影响因子：
7.4
通讯作者：
Dai, Jiale
中科院分区：
文献类型：
--
作者：
Pan, Yong;Wang, Lei;Dai, Jiale
Na+/H+交换调节因子1(NHERF1，又称EBP50或NHERF)是一种在乳腺癌中可能存在的肿瘤抑制基因。NHERF1位于17q25.1，在乳腺肿瘤发生过程中经常成为靶点。超过50%的乳腺肿瘤存在NHERF1基因的杂合性缺失(LOH)。此外，NHERF1在原发乳腺癌和乳腺癌细胞系的子集中发生突变。NHERF1基因的杂合性缺失与乳腺肿瘤的侵袭性特征密切相关，这意味着NHERF1是一种单倍性不足的肿瘤抑制基因。然而，推测的NHERF1肿瘤抑制活性尚未得到功能验证。为了证实我们的基因分析所表明的NHERF1的肿瘤抑制活性，我们使用逆转录病毒转导的短发夹状RNA(ShRNA)在乳腺癌细胞MCF7和T47D中下调了NHERF1的表达。然后评估这些细胞在体外和体内的细胞生长情况。对照组和NHERF1基因敲除的细胞也被血清饥饿并重新喂养，以通过荧光激活的细胞分类分析来比较它们的细胞周期进程。我们发现，与载体对照细胞相比，内源性NHERF1在T47D或MCF7细胞中的下调导致在锚定依赖和非锚定条件下的细胞增殖增强。将NHERF1基因敲除的T47D细胞植入无瘤小鼠的乳房脂肪垫，形成了比对照细胞更大的肿瘤。我们发现，血清饥饿的NHERF1基因敲除细胞在血清再刺激后比对照细胞更快地从G1过渡到S。免疫印迹分析显示，NHERF1基因敲除细胞周期进程加快的同时伴随着细胞周期蛋白E表达的增加和Rb磷酸化水平的升高。我们的研究结果表明，乳腺上皮细胞中正常的NHERF1功能涉及细胞周期进程的阻断。本研究肯定了NHERF1在乳腺中的抑瘤活性，这可能与其对细胞周期的调节作用有关。这一新的肿瘤抑制途径在人类乳腺癌中值得进一步研究，这可能会带来治疗机会。
Na+/H+ exchanger regulatory factor 1 (NHERF1, also known as EBP50 or NHERF) is a putative tumour suppressor gene in human breast cancer. Located at 17q25.1, NHERF1 is frequently targeted during breast tumourigenesis. Loss of heterozygosity (LOH) at the NHERF1 locus is found in more than 50% of breast tumours. In addition, NHERF1 is mutated in a subset of primary breast tumours and breast cancer cell lines. LOH at the NHERF1 locus is strongly associated with aggressive features of breast tumours, implicating NHERF1 as a haploinsufficiency tumour suppressor gene. However, the putative NHERF1 tumour suppressor activity has not been functionally verified. To confirm the NHERF1 tumour suppressor activity suggested by our genetic analyses, we used retrovirus-transduced short hairpin RNA (shRNA) to knock down NHERF1 expression in breast cancer cell lines MCF7 and T47D. These cells were then assessed for cell growth in vitro and in vivo. The control and NHERF1 knockdown cells were also serum-starved and re-fed to compare their cell cycle progression as measured by fluorescence-activated cell sorting analyses. We found that downregulation of the endogenous NHERF1 in T47D or MCF7 cells resulted in enhanced cell proliferation in both anchorage-dependent and -independent conditions compared with that of the vector control cells. NHERF1 knockdown T47D cells implanted at mammary fat pads of athymic mice formed larger tumours than did control cells. We found that serum-starved NHERF1 knockdown cells had a faster G1-to-S transition after serum re-stimulation than the control cells. Immunoblotting showed that the accelerated cell cycle progression in NHERF1 knockdown cells was accompanied by increased expression of cyclin E and elevated Rb phosphorylation level. Our findings suggested that the normal NHERF1 function in mammary epithelial cells involves blockage of cell cycle progression. Our study affirmed the tumour suppressor activity of NHERF1 in breast which may be related to its regulatory effect on cell cycle. It warrants future investigation of this novel tumour suppressor pathway in human breast cancer which may turn up therapeutic opportunities.
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影响因子：
4.8
作者：
He, JQ;Lau, AG;Hall, RA
通讯作者：
Hall, RA
影响因子：
11.4
作者：
MITTNACHT, S;LEES, JA;WEINBERG, RA
通讯作者：
WEINBERG, RA
DOI：
10.1073/pnas.031538898
发表时间：
2001-01-30
影响因子：
11.1
作者：
Raghuram, V;Mak, DOD;Foskett, JK
通讯作者：
Foskett, JK
影响因子：
4
作者：
James, MF;Beauchamp, RL;Ramesh, V
通讯作者：
Ramesh, V
影响因子：
7.8
作者：
Reczek, D;Berryman, M;Bretscher, A
通讯作者：
Bretscher, A