《Interferon-alpha-2b induces autophagy in hepatocellular carcinoma cells through Beclinl pathway.》.pdf

CancerBiolMed2014；11：64—68．doi：10．7497／j．issn．2095—3941．2014．01．006 ORIGINALARTICLE Interferon-alpha-2binducesautophagyinhepatoce¨ular carcinomacellsthroughBeclinlpathway JunZhao一，Ming—LiWang，ZengLi，Dong—MeGao，YuCai。JunChang，Shi—PingWang ， DepartmentofParasitology,XiangyaMedicalSchool，CentralSouthUniversity,Changsha410078，China；Departmentof Microbiology,AnhuiMedicalUniversity,Hefei230032，China；DepartmentofClinicalLaboratory,theThirdAffiliatedHospital ofAnhuiMedicalUniversity,Hefei230032，China；DepartmentofOrthopedics，TheFirstAffiliatedHospitalofAnhuiMedical University,Hefei230032，China Objective：TodeterminewhetherInterferon—alpha-2b(IFN—a2b)canmodulatetheautophagicresponseinhepatocellular carcinomacells． Methods：HepatocellularcarcinomacellsweretreatedwithIFN—a2b．Autophagywasassessedbyacridineorangestaining， GFP—LC3dottedassa~transmissionelectronmicroscopyandimmunoblotting． Results：AcridineorangestainingshowedthatIFN-a2btriggeredtheaccumulationofacidicvesicularandautolysosomes inHepG2cells．TheacridineorangeHepG2cellratioswere(4．3+1．o)％，(6．9±1．4)％，and(13．1±2．3)％respectively,after treatmentwith100，1,000，and10J000IU／mLIFN-a2bfor48h．AmarkedlypunctatepatternwasobservedinHepG2 cellstreatedwith10，000IU／mLIFN—a2bofr48h，butonlydifluseandweaklyfluorescentGFP-LC3punctawasobserved incontrolcells．HepG2cellstreatedwith10J000IU／mLIFN·a2bfor48hdevelopedautophagosome—likecharacteristicsJ includingsingle—ordouble—membranevacuolescontainingintactanddegradedcellulardebris．TheBeclinlandLC3一II proteinexpressionwasup—regulatedbyIFN—a2btreatment． Conclusion：Autophagycanbeinducedinadose—dependentmannerbytreatmentwithI