Klotho inhibits growth and promotes apoptosis in human lung cancer cell line A549.
Klotho inhibits growth and promotes apoptosis in human lung cancer cell line A549.
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Klotho 抑制人肺癌细胞系 A549 生长并促进细胞凋亡
DOI：
10.1186/1756-9966-29-99
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发表时间：
2010-07-19
期刊：
影响因子：
--
通讯作者：
Wu J
中科院分区：
文献类型：
--
作者：
Chen B;Wang X;Zhao W;Wu J
背景Klotho作为一种新的抗衰老基因，可以进入血液循环，作为一种多功能的体液因子影响多种生物过程。最近发表的研究表明，klotho也可以作为一种潜在的肿瘤抑制因子。本研究旨在探讨Klotho对人肺癌细胞株A549的作用及其可能的作用机制。方法构建编码Klotho或Klotho特异性shRNA的质粒，在体外过表达或敲除Klotho。分别用pCMV6-MYC-KL和Klotho特异性shRNA处理A549细胞。用四甲基偶氮唑盐比色法检测Klotho对A549细胞的细胞毒作用，用流式细胞仪观察Klotho诱导A549细胞凋亡的情况。Western blotting检测pCMV6-MYC-KL或shRNAs对A549细胞IGF-1/胰岛素信号通路的激活作用。结果Klotho过表达抑制肺癌A549细胞增殖，Klotho沉默促进A549细胞增殖。Klotho对HEK-293细胞无明显影响。Klotho在A549细胞中的过度表达与IGF-1/胰岛素诱导的IGF-1R(IGF-1受体)/IR(胰岛素受体)的磷酸化降低相关(P<；0.01)。Klotho过表达可促进A549细胞的凋亡(P<；0.01)。Bcl家族基因bax的高表达上调，抗凋亡基因bcl2的表达下调(P<；0.01)。相反，shRNA沉默Klotho后，Bax和bcl2的表达分别下调(P<；0.05)和上调(P<；0.01)。结论Klotho能抑制A549细胞的增殖，促进其凋亡，其机制可能与抑制IGF-1/Ins途径有关，并参与调控凋亡相关基因bax/bcl2的表达。因此，Klotho在A549细胞中可以作为一种潜在的肿瘤抑制因子。
BackgroundKlotho, as a new anti-aging gene, can shed into circulation and act as a multi-functional humoral factor that influences multiple biological processes. Recently, published studies suggest that klotho can also serve as a potential tumor suppressor. The aim of this study is to investigate the effects and possible mechanisms of action of klotho in human lung cancer cell line A549.MethodsIn this study, plasmids encoding klotho or klotho specific shRNAs were constructed to overexpress or knockdown klotho in vitro. A549 cells were respectively treated with pCMV6-MYC-KL or klotho specific shRNAs. The MTT assay was used to evaluate the cytotoxic effects of klotho and flow cytometry was utilized to observe and detect the apoptosis of A549 cells induced by klotho. The activation of IGF-1/insulin signal pathways in A549 cells treated by pCMV6-MYC-KL or shRNAs were evaluated by western blotting. The expression levels of bcl-2 and bax transcripts were evaluated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).ResultsOverexpression of klotho reduced the proliferation of lung cancer A549 cells, whereas klotho silencing in A549 cells enhanced proliferation. Klotho did not show any effects on HEK-293 cells. Klotho overexpression in A549 cells was associated with reduced IGF-1/insulin-induced phosphorylation of IGF-1R (IGF-1 receptor)/IR (insulin receptor) (P< 0.01). Overexpression of klotho can promote the apoptosis of A549 cells (P< 0.01). Overexpression of klotho, a bcl family gene bax, was found up-regulated and bcl-2, an anti-apoptosis gene, was found down-regulated (P< 0.01). In contrast, bax and bcl-2 were found down-regulated (P< 0.05) and up-regulated (P< 0.01), respectively when silencing klotho using shRNAs.ConclusionsKlotho can inhibit proliferation and increase apoptosis of A549 cells, this may be partly due to the inhibition of IGF-1/insulin pathways and involving regulating the expression of the apoptosis-related genes bax/bcl-2. Thus, klotho can serve as a potential tumor suppressor in A549 cells.
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影响因子：
8.3
作者：
Ma Z;Dong A;Kong M;Qian J
通讯作者：
Qian J
影响因子：
254.7
作者：
Jemal, Ahmedin;Siegel, Rebecca;Thun, Michael J.
通讯作者：
Thun, Michael J.
影响因子：
8.3
作者：
Mitani, H;Ishizaka, N;Nagai, R
通讯作者：
Nagai, R
DOI：
10.1006/bbrc.1997.8019
发表时间：
1998-01-26
影响因子：
3.1
作者：
Matsumura, Y;Aizawa, H;Nabeshima, Y
通讯作者：
Nabeshima, Y
影响因子：
5.6
作者：
Mattarocci, Stefano;Abbruzzese, Claudia;Paggi, Marco G.
通讯作者：
Paggi, Marco G.