首页> 外文期刊>Biochimica et Biophysica Acta. General Subjects >Orai1/CRACM1 overexpression suppresses cell proliferation via attenuation of the store-operated calcium influx-mediated signalling pathway in A549 lung cancer cells.
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Orai1/CRACM1 overexpression suppresses cell proliferation via attenuation of the store-operated calcium influx-mediated signalling pathway in A549 lung cancer cells.

机译:Orai1 / CRACM1的过表达通过减弱A549肺癌细胞中的钙离子介导的信号通路来抑制细胞增殖。

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BACKGROUND: Orai1/CRACM1 is a principal component of the store-operated calcium channels. Store-operated calcium influx is highly correlated with inflammatory reactions, immunological regulation, and cell proliferation. Epidermal growth factor (EGF), which plays an important role in the regulation of cell proliferation, can activate store-operated calcium channels. However, the consequences of Orai1/CRACM1 overexpression in EGF-mediated lung cancer cells growth are not known. METHODS: To investigate the role of Orai1/CRACM1 in EGF-mediated lung cancer cell proliferation, Orai1/CRACM1 plasmids were transfected into cells by lipofection. A cell proliferation assay, immunofluorescence staining, flow cytometry, and real-time polymerase chain reaction were employed to monitor cell proliferation. The calcium influx signals were investigated using a fluorescent-based calcium assay. RESULTS: Transfection of Orai1/CRACM1 plasmids resulted in the inhibition of EGF-mediated cell proliferation. ERK1/2 and Akt phosphorylation were inhibited by Orai1/CRACM1 overexpression. Expression of the cell cycle modulator p21 was induced in the Orai1/CRACM1-overexpressing cells, whereas the expression of cyclin D3 was reduced. Flow cytometry revealed that overexpression of Orai1/CRACM1 resulted in G0/G1 cell cycle arrest. Importantly, Orai1/CRACM1 overexpression significantly attenuated EGF-mediated store-operated calcium influx. In addition, application of 2-APB, a store-operated calcium channel inhibitor, resulted in the inhibition of EGF-mediated cancer cell proliferation. CONCLUSIONS: We conclude that Orai1/CRACM1 overexpression attenuates store-operated Ca(2+) influx that in turn blocks EGF-mediated proliferative signaling and drives cell cycle arrest.
机译:背景:Orai1 / CRACM1是商店运营的钙通道的主要组成部分。商店运营的钙流入与炎症反应,免疫调节和细胞增殖高度相关。在调节细胞增殖中起重要作用的表皮生长因子(EGF)可以激活钙存储通道。但是,Orai1 / CRACM1过表达在EGF介导的肺癌细胞生长中的后果尚不清楚。方法:为了研究Orai1 / CRACM1在EGF介导的肺癌细胞增殖中的作用,通过脂质转染将Orai1 / CRACM1质粒转染到细胞中。细胞增殖测定,免疫荧光染色,流式细胞仪和实时聚合酶链反应用于监测细胞增殖。使用基于荧光的钙测定法研究钙流入信号。结果:转染Orai1 / CRACM1质粒可抑制EGF介导的细胞增殖。 Orai1 / CRACM1过表达抑制ERK1 / 2和Akt磷酸化。在过表达Orai1 / CRACM1的细胞中诱导细胞周期调节剂p21的表达,而细胞周期蛋白D3的表达降低。流式细胞仪显示,Orai1 / CRACM1的过度表达导致G0 / G1细胞周期停滞。重要的是,Orai1 / CRACM1的过表达显着减弱了EGF介导的存储操纵性钙内流。此外,应用2-APB(一种由商店运营的钙通道抑制剂)可抑制EGF介导的癌细胞增殖。结论:我们得出结论,Orai1 / CRACM1过表达减弱了存储操作的Ca(2+)内流,进而阻止了EGF介导的增殖信号并驱动细胞周期停滞。

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