Welcome to visit Acta Anatomica Sinica! Today is
Chinese
Expressions and implications of end-binding protein 1 in autophagic Siha cells
LIU Yi-qun LUO Xiong-yan JIANG Hong XIA Yan-hui LIU Qing-song*
Acta Anatomica Sinica ›› 2014 ›› Issue (2) : 228-233.
Expressions and implications of end-binding protein 1 in autophagic Siha cells
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 colchicinein vitro. Specific green fluoresent protein(GFP)-LC3 was then be detected to confirm the autophagosome formation by fluorescence microscopy withFlowCellectTMGFP-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.
［1］Sun Y, Liu JH, Jin L, et al. Over-expression of the Beclin1 gene upregulates chemosensitivity to anti-cancer drugs by enhancing therapy-induced apoptosis in cervix squamous carcinoma CaSki cells ［J］. Cancer Lett, 2010, 294(2): 204-210.
［2］K ?chl R, Hu XW, Chan EY, et al. Microtubules facilitate autophagosome formation and fusion of autophagosomes with endosomes ［J］. Traffic, 2006, 7(2): 129-145.
［3］Bian ML. WHO(2006)practice guidelines for cervical cancer prevention［J］. Chinese Journal of Practical Gynecology and Obostetrics, 2007, 23(7):557-560. (in Chinese)
卞美璐. WHO (2006年) 宫颈癌综合防治实践指南简介［J］. 中国实用妇科与产科杂志, 2007, 23(7): 557-560.
［4］Chunrong H, Baocun S, Wei W, et al. Overexpression of microtubuleassociated protein-1 light chain 3 is associated with melanoma metastasisand vasculogenic mimicry ［J］. Tohoku J Exp Med, 2011, 223(22): 243-251.
［5］Duran A, Amanchy R, Linares JF, et al. p62 is a key regulator of nutrient sensing in the mTORC1 pathway ［J］. Mol Cell, 2011, 44 (1):134-146.
［6］Kim DH, Sarbassov DD, Ali SM, et al. mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery［J］. Cell, 2002, 110 (2): 163-175.［7］Bjorkoy G, Lamark T, Brech A, et al. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death ［J］. J Cell Biol, 2005, 171(4): 603-614.
［8］Moscat J, Diaz-Meco MT. Feedback on fat: p62-mTORC1-autophagy connections ［J］. Cell, 2011, 147(4): 724-727.
［9］Puissant A, Fenouille N, Auberger P. When autophagy meets cancer through p62/SQSTM1 ［J］. Am J Cancer Res, 2012, 2(4): 397-413.
［10］Wang Y, Singh R, Xiang Y, et al. Macroautophagy and chaperone-mediated autophagy are required for hepatocyte resistance to oxidant stress ［J］. Hepatology, 2010, 52(1): 266-277.
［11］Jahreiss L, Menzies FM, Rubinsztein DC. The itinerary of autophagosomes: from peripheral formationto kiss-and-run fusion with lysosomes ［J］. Traffic, 2008, 9(4): 574-587.［12］Aplin A, Jasionowski T, Tuttle DL, et al. Cytoskeletal elements are required for the formation and maturation of autophagic vacuoles ［J］. J Cell Physiol, 1992, 152(3): 458-466. ［13］Seglen PO, Berg TO, Blankson H, et al. Structural aspects of autophagy ［J］. Adv Exp Med Biol, 1996, 389: 103-111.
［14］Monastyrska I, Rieter E, Klionsky DJ, et al. Multiple roles of the cytoskeleton in autophagy ［J］. Biol Rev Camb Philos Soc, 2009, 84(3): 431-448.
［15］Orsi A, Polson HE, Tooze SA. Membrane trafficking events that partake in autophagy ［J］. Curr Opin Cell Biol, 2010, 22(2):150-156.
［16］Fass E, Shvets E, Degani I, et al. Microtubules support production of starvation-induced autophagosomes but not their targeting and fusion with lysosomes［J］. J Biol Chem, 2006, 281(47): 36303-36316.
［17］K ?chl R, Hu XW, Chan EY, et al. Microtubules facilitate autophagosome formation and fusion of autophagosomes with endosomes ［J］. Traffic, 2006, 7(2):129-145.
［18］Xie R, Nguyen S, McKeehan K, et al. Microtubule-associated protein 1S (MAP1S) bridges autophagic components with microtubules and mitochondria to affect autophagosomal biogenesis and degradation ［J］. J Biol Chem, 2011, 286(12): 10367-10377.
［19］Jánosi IM, Chrétien D, Flyvbjerg H. Structural microtubule Cap: stability, catastrophe, rescue, and third state ［J］. Biophys J, 2002, 83(3): 1317-1330.
［20］Jiang K, Akhmanova A. Microtubule tip-interacting proteins: a view from both ends ［J］. Curr Opin Cell Biol, 2011, 23(1):94-101.
［21］Tirnauer JS, Bierer BE. EB1 proteins regulate microtubule dynamics, cell polarity, and chromosome stability ［J］. J Cell Bio, 2000, 149(4) 761-766.
［22］Ligon LA, Shelly SS, Tokito M, et al. The microtubule plus-end proteins EB1 and dynactin have differential effects on microtubule polymerization ［J］. Mol Biol Cell, 2003, 14(4): 1405-1417.
［23］Wang YH, Zhou XB, Zhu H, et al. Overexpression of EB1 inhuman esophageal squamous cell carcinoma (ESCC) may promote cellular growth by activating beta-catenin/TCF pathway ［J］. Oncogene, 2005, 24(44): 6637-6645.
［24］Kaplan KB, Burds AA, Swealow JR, et al. A role for the adenomatous polyposis coli protein in chromosome segregation ［J］. Nat Cell Bio, 2001, 3(4): 429-432.
/
| 〈 |
|
〉 |
