489 receptorα,erα) ^;% : #!"#$%&(epthelialgrowthfactor, EGF) ;+, ; [3]!"#$%&>? X` < < M N O < _ < = >? (estrogenreceptor, ER) [4], ();+, O,EGFX` C

2017 4 37 4 :1001 6325(2017)04 0488 05 Basic& ClinicalMedicine April2017 Vol.37 No.4 研究论文 1, 1, 2, 1, 2 (1 -./01, -. 266003;2 /012345 67,23 100050) :! 89!"#$%&(EGF):& ()* Ishikawa+,;, <=>? α(erα)@ Ack1ABCDE ; "# <= FG,EGF H Ishikawa(),CCK 8IJK& ()+,;WesternblotJK() ERαL Ack1MNO; N< PQRST()U,JK Ishikawa()+,@ ERαL Ack1MNOVW $% EGFX+Y Ishikawa()+,(P<0 05), ' ERαTyr 537 MNO@ Ack1MNO; PQRU,()+, GZ(P<0 05),ERαTyr 537 MNO@ Ack1MNO[\GC $& EGF' Ishikawa()+,,BE X ]^ ERαTyr 537 MNO@<_ Ack1< `abc ' :& ;!"#$%&; <=>?;Ack1; PQR;MNO :R711 74 ()*+:A Epidermalgrowthfactorpromotes proliferationofendometrialadenocarcinomacellineishikawa ZHANGJing 1,TIANTian 1,LIUJing 2,CUIZhu mei 1,LIUYuan bo 2 (1 Dept.ofGynecology,theafiliatedHospitalofQingdaoUniversity,Qingdao266003; 2 Dept.ofHematology,BeijingTiantanHospital,CapitalMedicalUniversity,Beijing100050,China) Abstract:Objective Toinvestigatetheefectofepidermalgrowthfactor(EGF)ontheproliferationofendometrial adenocarcinomacels,phosphorylationofestrogenreceptorα(erα)andack1intheabsenceofestrogen.methods IshikawacellinewasstimulatedbyEGFwithoutestrogensetings,CelCountingKit 8(CCK 8)wasusedtoeval uatecelproliferation,westernblotwasusedtodetecterαphosphorylationandack1phosphorylation.giving tyrosineinhibitordasatinibtoasestheefectofegfoncelproliferation,phosphorylationoferαandack1inish ikawacels.results EGFenhancedtheproliferationofendometrialadenocarcinomacels(P<0 05).EGFin ducederαphosphorylationattyr 537andphosphorylationofAck1.Comparedwithuntreatedcontrol,Dasatinib inhibitedtheproliferationofendometrialadenocarcinomacels(p<0 05),phosphorylationofERαTyr 537and Ack1.Conclusions EGFpromotesIshikawacelsproliferationintheposiblewayofactivatingERαsite specific phosphorylationattyr 537andphosphorylationAck1,whichcouldbeblockedbydasatinib. Keywords:endometrialneoplasms;epidermalgrowthfactor;estrogenreceptor;Ack1;dasatinib;phosphorylation & ; #,*, H&?;, #, *,B H & A ; O [1 2], E 89 & # <=bc,b <=>? α(estrogen :2016 12 05 :2017 01 11 : (81272842) (correspondingauthor):cuizhumei1966@126.com;yuanbol@ccmu.edu.cn

489 receptorα,erα) ^;% : #!"#$%&(epthelialgrowthfactor, EGF) ;+, O@6 ; [3]!"#$%&>? X` < < M N O < _ < = >? (estrogenreceptor, ER) [4], ();+, O,EGFX` CD Ack1]^ < =>?(androgenreceptor,ar) MNO,' ()+, [5] EGFA& :()+,; L ce c 89 & ()* Ishikawa 89 :, A <= G,EGF:& ()+,;, 89B Ack1@ ERαMNO;c*, EGF:& ( )+,; L ccde 1,-."# 1 1,- 1 1 1 (): <= & ()* Ishikawa(23 @() ) 1 1 2 @?:EGF(R&D );Ack1? p Ack1?L ER? (Abcam Biotechnology );ERTyr 537 MNO?(SantaCruz ); P Q R (CelSignaling Technology ); L O ; (2 3 b ); CCK 8 (CelCountingKit 8,23 O 89) 1 2 "# 1 2 1 ( ) L S T: 10% 6 100U/mL- = 100U/mL =; DMEM F/ 12 7 (),H 37 CO 2?! 5%; ( ) " # ( ) $ % & 90% ~ 100%U,0 25%' O 1 3; +, b () SH:! +,;() EGF(100μg/L)ST, E2(β estradiol,10μg/l)st, PQR(dasat inib)(10nmol/l~800nmol/l)st @:* 1 2 2 CCK 8JK()+,:+:!+,; Ishikawa(),, 0 25%' OU-. ()/7,0 10 4 1 / ; Ishikawa() H 96, 2 EGF@ PQR 24hU, 3* CCK 84 5 6JK()+,78 1 2 3 WesternblotJK ERTyr 537 MNOL Ack1MNO:A 6 Ishikawa(),#( )+,9:;, PQR ST Ishikawa() 2hU, 2 EGFST 1hU, ; PBS ( ) 3, <=> 200μLRIPA()?@7,HA B 20min,CDE (), ()?@,4 G 12000r/min 15min, +B 7, BCAI F G,+ 40μg H I,` 10%J KLNM NO P,300mA Q G 2h,RSTU VWX 2hU,= > Ack1 p Ack1 ERp Y537L ER? 4 Y 3,15min/,?RSY 1h, 3,15min/,O I Image J Z,` MNO[; \MNO [; ]; &^ ^ [; ; : G 1 3 /01 _ SPSS19 0 `Z!a,`G bc% W,_ (! ± (x±s)!, _ H tj 2 $% 2 1 234,EGF56789:; EGF(10μg/L)@ E2(10μg/L)ST; Ishikawa () &^( H:* (P<0 05)( 1) P<0 05comparedwithcontrol 1 234,EGF78 Ishikawa Fig1 EGFpromotestheproliferationofIshikawacels intheabsenceofestrogen(x±s,n=9)

490 Basic&ClinicalMedicine 2017 37(4) P<0 05comparedwithcontrol; # P<0 05comparedwithdasatinib:0&EGF:100 2 <=>?@ABCDE EGF! Ishikawa Fig2 DasatinibinhibitedtheproliferationofIshikawacelsinducedbyEGF(x±s,n=9) 2 2 < = >? EGF! Ishikawa PQR(10nmol/L~800nmol/L) EGF]^; Ishikawa()+,, PQR & +=,() &^ Z (P<0 05)( 2) 2 3 EGF Ack1 FGH ERIJKL FG EGF]^,Ishikawa()MNO(<_O VW); Ack1@ Tyr 537ER[\( 2 100μg/LEGFST Ishikawa(), :*, Ack1@ ERTyr 537 ;MNO[\ ( 3,! 1) 2 4 <=>? EGF! ERTyr 537IJ KL FG PQRX EGF]^; ERTyr 537 MNO, G+=,ER Ack1 M N 1 EGFO Ishikawa Ack1 p Ack1 ER H ERTyr 537 N<! Table1 EfectsofEGFonexpresionsofAck1,p Ack1, ERandERTyr 537proteininIshikawacels (x±s,n=9) group p Ack1/Ack1 p ER/ER control 0 0114±0 0112 0 0803±0 0072 EGF 0 5199±0 0347 0 9721±0 0333 P<0 05comparedwithcontrolgroup. NO[\ Z, PQR &/H 400nmol/L,Ack1L ERTyr 537 MNO ( 4,! 2) N 2 DasatinibO Ishikawa Ack1 p Ack1 ERH ERTyr 537 N<! Table2 EfectsofDasatinibonexpresionsofAck1, p Ack1,ERandERTyr 537proteinin Ishikawacels(x±s,n=9) group p Ack1/Ack1 p ER/ER control 0 0000±0 0000 0 0000±0 0000 0nmol/L 0 3558±0 0783 0 4044±0 0616 10nmol/L 0 2998±0 0650 0 2064±0 0338 50nmol/L 0 3300±0 0713 0 0613±0 0147 # 100nmol/L 0 3067±0 0675 0 0693±0 0207 # 200nmol/L 0 0695±0 0152 # 0 0833±0 0109 # 3 EGF Ishikawa M Ack1H ER Tyr 537IJKL FG Fig3 EGFinducedAck1andTyr 537ER phosphorylationofishikawacellines 400nmol/L 0 1120±0 0032 # 0 0141±0 0031 # 800nmol/L 0 0000±0 0000 # 0 0000±0 0000 # P<0 05comparedwithcontrolgroup; # P<0 05comparedwithDasat inib:0&egf:100group.

491 4 <=>? Ishikawa M EGF! Ack1H ER FG Fig4 DasatinibinhibitedEGFinducedphosphorylationofAck1andER 3 & <= >? %<_& () C %;!, # EGFLB>? ' ()+, [6] EGF <=;, ` ;, B>? %U ();+, OL _ O [7] `? EGF:& () Ishikawa;+,', X ; c &E EGF!"#$%&>? %UX` ]^ NMNO, < N< `a,' () [8] 89,A? < = FG,() Ack1< @ Src< ^; AR NMNO AR_O;c,X]^ [5] < = G EGF:& (); @ ce!,egfa <= GX' & Ishikawa;+,,E X EGF]^ ERαTyr 537 MNO@ N< Ack1 MNObc N< PQR G ; EGF]^; ERαTyr 537 MNO@ Ack1MNO[\,^ Ishikawa ()+, GZ, C :H Ack1 B & CD ERα Tyr 537 ;_O, 89 N< ;() ^` a,a();+,@ O b, O MN B;MN ; N B, B #MNO [9] N< ();, ()+,, ()+,@ #6 [10] PQR N<,` CD Bcr Abl@ Src<? ;#$ [11], 89; b89!, PQRX ();+, @ [12], X ( );+,@ [13] 89, PQRGC ARTyr 267@ ARTyr 534 MNO [\, PQRA ; a [14] 89 PQRA& () X ERαTyr 537 @ Ack1 < MNO, PQRA& ; ; a P (: [1]WongYF,CheungTH,LoKW,etal.Identificationofmo lecularmarkersandsignalingpathwayinendometrialcancer inhongkongchinesewomenbygenome widegeneexpres sionprofiling[j].oncogene,2007,26:1971 1982. [2]ZhangY,LiuZ,YuX,etal.Theasociationbetweenmet abolicabnormalityandendometrialcancer:alargecase controlstudyinchina[j].gynecoloncol,2010,117: 4 46. [3]EjskjaerK,SorensenBS,PoulsenSS,etal.Expresionof theepidermalgrowthfactorsysteminendometrioidendome trialcancer[j].gynecoloncol,2007,104:158 167. [4]BritonDJ,HutchesonIR,KnowldenJM,etal.Bidirec

492 Basic&ClinicalMedicine 2017 37(4) tionalcrostalkbetweenerαandegfr signalingpath waysregulatestamoxifen resistantgrowth[j].breastcanc errestreat,2006,96:131 146. [5],,,. <= G!"#$% &]^ *)*+,L <=>?MNO[J].,2014,34:305 309. [6],,. PTEN: EGF]^ ;& () ERK_O; [J].() &,2002,18:369 372. [7].!"#$%& @# [J].,2006,18:14 17. [8]SeshacharyuluP,PonnusamyMP,HaridasD,etal.Targe tingtheegfr signalingpathwayincancertherapy[j]. ExpertOpinTherTargets,2012,16:15 31. [9]TraxlerP.Tyrosinekinasesastargetsincancertherapy succesesand failures[j]. ExpertOpin TherTargets, 2003,7:215 34. [10] NormannoN,DeLucaA,BiancoC,etal.Epidermal growthfactorreceptor(egfr)signalingincancer[j]. Gene,2006,366:2 17. [11]LeXF,MaoW,LuZ,etal.Dasatinibinducesautophag icceldeathinhumanovariancancer[j].cancer,2010, 116:4980 4990. [12]PichotCS,HartiqSM,XiaL,etal.Dasatinibsynergizes withdoxorubicintoblockgrowth,migration,andinvasion ofbreastcancercels[j].brjcancer,2009,101: 38 47. [13],. PQR%: ()+,@ ; [J].,2011,26:3784 3787. [14],,,. PQR () <=>? M N O E 8 9 [J]. 8 9, 2016,28:361 365. 新闻点击 QRS TUVWX 据英国 BBC 新闻 (BBCNEWS)2015 11 24 报道, 有研究指出, 孤单会抑制免疫系统, 让人更容易罹患疾病, 危害程度是肥胖的 2 倍 美国芝加哥大学 (UniversityofChicago) 研究人员发现, 在晚年觉得最为孤单的群体中, 早死风险比那些觉得最被需要的群体高出近 15% 研究称, 孤单危害健康的程度是肥胖的 2 倍 研究团队说, 远离人群会弱化人体抗病毒的能力, 并让血压升高到心脏病发作或脑卒中的危险值, 并增加忧郁症风险 研究团队发现, 免疫系统基因 CTRA 在 50 岁以上 感到非常寂寞的群体身上特别活跃, 这会抑制对抗病毒的免疫反应, 并且增加炎性反应, 这与心脏疾病和虚弱等许多健康问题有关 研究人员 JohnCacioppo 认为, 觉得寂寞的人可以通过公益活动来交友, 如念书给盲友听, 或是指导儿童的运动队等 另外, 养宠物作伴也会有帮助, 他发现养狗会降低寂寞感, 但养猫 蜥蜴或仓鼠就没有这种效果 该项研究发表在 美国国家科学院院报 (PNAS) Y T ZWX 据美国 WebMD 医学新闻网 (2015 11 26) 报道, 研究人员认为, 长期住在嘈杂路边的人可能会增加罹患忧郁症的风险 相较于住家附近较少交通噪声的人来说, 住在交通繁忙区域的人罹患忧郁症的风险提高 25% 然而, 德国的研究人员发现, 这种风险大部分局限在穷人 失业者 教育程度低 吸烟或是失眠的人 Esen 大学的研究人员 EsterOrban 认为, 虽然他们不能确认, 但认为噪声会造成压力与烦恼 如果噪声持续很长一段时间, 且很大声的话, 可能会造成忧郁症 如果觉得马路上的噪声很大, 可以使用耳塞, 如果噪声会干扰睡眠, 卧室最好远离繁忙交通的路段 这篇报告刊登在 2015 11 25 环境健康展望 (EnvironmentalHealthPerspectives)