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Molecular imaging of low-power laser irradiation induced cell proliferation

机译:低功率激光辐射诱导细胞增殖的分子成像

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Low-power laser irradiation (LPLI) has been shown to promote cell proliferation in various cell types, yet the mechanism of which has not been fully clarified. Studying the signaling pathways involved in the laser irradiation is important for understanding these processes. The Ras/Raf/MEK/ERK (extracellular-signal-regulated kinase) signaling pathway is a network that governs proliferation, differentiation and cell survival. Recent studies suggest that Ras/Raf signaling pathway is involved in the LPLI-induced cell proliferation. Protein kinase Cs (PKCs) have been recently presumed to be involved in the regulation of cell proliferation induced by LPLI. In present study, to monitor the direct interaction between Ras and Raf and PKCs activation after LPLI treatment in living cells in real time, Raichu-Ras reporter and C kinase activity reporter (CKAR) were utilized, both of which were constructed based on fluorescence resonance energy transfer (FRET) technique. Our results show that the direct interaction between Ras and Raf is monitored during cell proliferation induced by LPLI (0.8 J/cm2) in serum-starved human lung adenocarcinoma cells (ASTC-a-1) expressing Raichu-Ras reporter using FRET imaging on laser scanning confocal microscope, and that the increasing dynamics of PKCs activity is also monitored during cell proliferation induced by LPLI (0.8 J/cm2) in serum-starved ASTC-a-1 cells expressing CKAR reporter using the similar way. Taken together, LPLI induces the ASTC-a-1 cell proliferation by activated Ras directly interacting with Raf and by specifically activating PKCs.
机译:已经显示低功率激光照射(LPLI)以促进各种细胞类型的细胞增殖,但其机制尚未完全澄清。研究激光照射中所涉及的信令途径对于理解这些过程非常重要。 RAS / RAF / MEK / ERK(细胞外信号调节激酶)信号传导途径是一种治理增殖,分化和细胞存活的网络。最近的研究表明RAS / RAF信号通路涉及LPLI诱导的细胞增殖。最近推测蛋白激酶Cs(PKC)参与LPLI诱导的细胞增殖调节。在目前的研究中,在实时的LPLI处理后,监测RAS和RAF与PKCS活化后的直接相互作用,利用Raichu-Ras报道和C激酶活性报告(CKAR),两者都基于荧光共振构建能量转移(FRET)技术。我们的研究结果表明,使用FRET成像在激光上使用FRET成像在表达RACH-RAS记者的LPLI(0.8J / cm2)在血清饥饿的人肺腺癌细胞(ASTC-A-1)中诱导的细胞增殖期间进行直接相互作用扫描共聚焦显微镜,并且在使用类似方式表达CKAR记者的血清饥饿ASTC-A-1细胞中的LPL(0.8J / cm2)诱导的细胞增殖期间也监测PKCS活性的增加动力学。将LPL一起携带,通过直接与RAF与特定激活PKC进行活化的Ras诱导ASTC-A-1细胞增殖。

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