Expressions and implications of end-binding protein 1 in autophagic Siha cells

doi:10.3969/j.issn.0529-1356.2014.02.015

AAS ›› 2014, Vol. ›› Issue (2): 228-233.doi: 10.3969/j.issn.0529-1356.2014.02.015

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Expressions and implications of end-binding protein 1 in autophagic Siha cells

LIU Yi-qun¹ LUO Xiong-yan³JIANG Hong³ XIA Yan-hui³ LIU Qing-song ^2,3*

1. Department of Respiratory Medicine, the Secondary Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China;2. Department of Laboratory Medicine, Northern Sichuan Medical College, Sichuan Nanchong 637000, China;3. Institute of Rheumatology and Immunology, Affiliated Hospital of Northern Sichuan Medical College, Sichuan Nanchong 637000, China)

Received:2013-08-19 Revised:2013-10-24 Online:2014-04-06 Published:2014-04-06
Contact: XIA Yan-hui LIU Qing-song E-mail:liuqingsong00@aliyun.com

Abstract

Abstract:

Objective To explore the expressions and implications of end-binding protein 1(EB1) in autophagic cervical cancer Siha cells. Method Real-time quantitative PCR and Western blotting were introduced to detect the mRNA and protein expressions of EB1, LC3 and p62 in Siha cells treated with Hank’s balanced salt solution (HBSS) or colchicinein vitro. Specific green fluoresent protein(GFP)-LC3 was then be detected to confirm the autophagosome formation by fluorescence microscopy withFlowCellectTMGFP-LC3 reporter autophagy assay kit. Result The mRNA and protein levels of LC3 and EB1 in cervical cancer Siha cells were increased significantly when prolong the duration of HBSS treatment (P<0.05). EB1 mRNA was positively correlated with LC3 mRNA expression (P<0.05). The expressions of p62 mRNA and protein reduced dramatically after HBSS treated (P<0.05), and was negatively related with the EB1 mRNA (P<0.05). The number of cells with GFP-LC3 and intensity of GFP-LC3 per cells were increased when starvation for 12 hours. However, the mRNA and protein levels of LC3, p62 and EB1 were not significantly changed when the cells treated with colchicine (P>0.05), and we cannot detected the fluoresce signaling of GFP-LC3 in cells at this moment. Conclusion The expression of EB1 in starved cells is dramatically increased, and accompanied with the increased expression of LC3 and autophagosome, decreased expression of p62. However, when microtubules with which EB1 interaction were destructed by colchicine, the above phenomenon disappears. All these results fully indicate the probably roles of EB1 in autophagy.

LIU Yi-qun LUO Xiong-yan JIANG Hong XIA Yan-hui LIU Qing-song*. Expressions and implications of end-binding protein 1 in autophagic Siha cells[J]. AAS, 2014, (2): 228-233.

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Expressions and implications of end-binding protein 1 in autophagic Siha cells

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