Anti-EGFR antibody efficiently and specifically inhibits human TSC2-/- smooth muscle cell proliferation. Possible treatment options for TSC and LAM.
Anti-EGFR antibody efficiently and specifically inhibits human TSC2-/- smooth muscle cell proliferation. Possible treatment options for TSC and LAM.
复制标题
DOI：
10.1371/journal.pone.0003558
复制
发表时间：
2008
期刊：
影响因子：
3.7
通讯作者：
Gorio A
中科院分区：
文献类型：
--
作者：
Lesma E;Grande V;Ancona S;Carelli S;Di Giulio AM;Gorio A
结节性硬化症（TSC）是一种由TSC1或TSC2基因突变引起的肿瘤综合征，其特征是错构瘤的发生。我们之前从一名TSC2患者的血管平滑肌脂肪瘤中分离出一群均匀的平滑肌样细胞（TSC2 - / - ASM细胞），这些细胞具有TSC2基因突变以及TSC2杂合性缺失（LOH），因此不产生TSC2基因产物tuberin。TSC2−/−ASM细胞增殖依赖于egf。观察EGF对转染TSC2基因的TSC2 - / - ASM细胞和TSC2 - / - ASM细胞增殖的影响。与TSC2−/−ASM细胞相比，转染TSC2的细胞的生长不依赖于EGF。此外，Akt、PTEN、Erk和S6的磷酸化水平明显降低。EGF是TSC2−/−ASM细胞的增殖因子。将TSC2−/−ASM细胞暴露于抗egfr抗体中，可显著抑制其增殖，恢复对HMB45抗体（TSC2−/−细胞表型标记物）的反应性，并抑制S6和ERK的组成磷酸化。将TSC2−/−ASM细胞暴露于雷帕霉素中可降低增殖率，但仅在电镀时添加雷帕霉素可降低增殖率。虽然雷帕霉素能有效抑制S6磷酸化，但在恢复HMB45反应性和阻断ERK磷酸化方面，雷帕霉素的效率低于抗egfr抗体。在TSC2−/−ASM细胞中，特异性PI3K抑制剂（如LY294002、wortmannin）和Akt1 siRNA对S6和ERK磷酸化的影响很小。转染tsc2基因后，观察Akt抑制剂敏感性。我们的研究结果表明，对于TSC - / - ASM细胞的生长和存活，EGF独立通路比涉及IGF-I的通路更重要，这种EGF依赖性是缺乏tuberin的结果。
Tuberous sclerosis complex (TSC), a tumor syndrome caused by mutations in TSC1 or TSC2 genes, is characterized by the development of hamartomas. We previously isolated, from an angiomyolipoma of a TSC2 patient, a homogenous population of smooth muscle-like cells (TSC2−/− ASM cells) that have a mutation in the TSC2 gene as well as TSC2 loss of heterozygosity (LOH) and consequently, do not produce the TSC2 gene product, tuberin. TSC2−/− ASM cell proliferation is EGF-dependent. Effects of EGF on proliferation of TSC2−/− ASM cells and TSC2−/− ASM cells transfected with TSC2 gene were determined. In contrast to TSC2−/− ASM cells, growth of TSC2-transfected cells was not dependent on EGF. Moreover, phosphorylation of Akt, PTEN, Erk and S6 was significantly decreased. EGF is a proliferative factor of TSC2−/− ASM cells. Exposure of TSC2−/− ASM cells to anti-EGFR antibodies significantly inhibited their proliferation, reverted reactivity to HMB45 antibody, a marker of TSC2−/− cell phenotype, and inhibited constitutive phosphorylation of S6 and ERK. Exposure of TSC2−/− ASM cells to rapamycin reduced the proliferation rate, but only when added at plating time. Although rapamycin efficiently inhibited S6 phosphorylation, it was less efficient than anti-EGFR antibody in reverting HMB45 reactivity and blocking ERK phosphorylation. In TSC2−/− ASM cells specific PI3K inhibitors (e.g. LY294002, wortmannin) and Akt1 siRNA had little effect on S6 and ERK phosphorylation. Following TSC2-gene transfection, Akt inhibitor sensitivity was observed. Our results show that an EGF independent pathway is more important than that involving IGF-I for growth and survival of TSC−/− ASM cells, and such EGF-dependency is the result of the lack of tuberin.
登录
查看更多内容
影响因子：
4.8
作者：
Harwood, Franklin C.;Shu, Lili;Houghton, Peter J.
通讯作者：
Houghton, Peter J.
影响因子：
6
作者：
Karbowniczek, M;Yu, J;Henske, EP
通讯作者：
Henske, EP
DOI：
10.1073/pnas.0405659101
发表时间：
2004-09-14
影响因子：
11.1
作者：
Roux, PP;Ballif, BA;Blenis, J
通讯作者：
Blenis, J
DOI：
10.1152/ajpgi.00403.2003
发表时间：
2004-03-01
影响因子：
4.5
作者：
Kuemmerle, JF;Zhou, HP;Bowers, JG
通讯作者：
Bowers, JG
影响因子：
3.6
作者：
Kenerson, H;Dundon, TA;Yeung, RS
通讯作者：
Yeung, RS