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1、CHINESEJOURNALOFANAT0MYVo1.37No.52014解剖學(xué)雜志2014年第37卷第5期褪黑素誘導(dǎo)人骨肉瘤MG63細(xì)胞凋亡黃宇林晉。周瑞祥。林建華陳宣維.(1福建醫(yī)科大學(xué)附屬第一醫(yī)院,2福建衛(wèi)生職業(yè)技術(shù)學(xué)院,3福建醫(yī)科大學(xué)人體解剖學(xué)與組織學(xué)胚胎學(xué)系,福州350004)摘要目的:觀察褪黑素對人骨肉瘤細(xì)胞系MG63增殖和凋亡的影響及其對細(xì)胞周期和Fas基因表達(dá)的調(diào)控。方法:體外培養(yǎng)MG63細(xì)胞,CCK8法檢測不同濃度褪黑素對MG63細(xì)胞增殖的影響,通過Hoechst33258染色觀察細(xì)胞凋亡的形態(tài)學(xué)變化,流式細(xì)胞術(shù)檢測細(xì)胞凋亡及細(xì)
2、胞周期改變,實(shí)時熒光定量PCR及免疫印跡檢測MG63細(xì)胞Fas基因表達(dá)情況。結(jié)果:CCK8法檢測顯示,褪黑素可抑制MG63細(xì)胞增殖,并具有時間和劑量依賴性;Hoechst33258染色可見凋亡小體;流式細(xì)胞術(shù)檢測顯示褪黑素可使凋亡的MG63細(xì)胞明顯增加,并使細(xì)胞周期阻滯在G2/M期;實(shí)時熒光定量PCR及免疫印跡分析顯示經(jīng)褪黑素作用后MG63細(xì)胞表達(dá)Fas蛋白明顯增強(qiáng)。結(jié)論:褪黑素對MG63細(xì)胞的增殖有抑制作用,并誘導(dǎo)其發(fā)生凋亡,其機(jī)制可能與細(xì)胞周期阻滯在G2/M期及上調(diào)Fas蛋白表達(dá)有關(guān)。關(guān)鍵詞褪黑素;骨肉瘤;Fas;細(xì)胞周期;實(shí)時熒光定量PCR
3、;免疫印跡MelatonininducesapoptosisinthehumanosteosarcomaMG63cells一HuangYu,LinJin2,ZhouRuixiang。,LinJianhua,ChenXuanwei△(1.FAffiliatedHospitalofFujianMedicalUniversity,2.FujJanHealthCollege,3.DepartmentofHumanAnatomy,HistologyandEmbryology,F(xiàn)ujianMedicalUniversity,F(xiàn)uzhou350004,chin
4、a)AbstractObjective:ToinvestigatetheeffectofmelatoninonproliferationandapoptosisinthehumanosteosarcomacelllineMG63andtheregulationonthecellcycleandFasgeneexpression.Methods:MG63cellswereculturedinvitroandtreatedwithdifferentconcentrationsofmelatonin.Cellproliferationwasdetect
5、edbyCCK8.Hoechst33258wasusedtodetectthemorphologicalchangeofapoptoticcells.MG63cellswereobservedbyflowcytometryforapoptosisandcellcycle.TheexpressionofFaswasobservedbyrealtimefluorescencequantitivePCRandWesternblotting.TheapoptoticbodywasobservedbyHoechst33258dyeing.Results:T
6、heproliferationofMG63cellswasinhibitedaftertreatment,whichexhibitedinadoseandtimedependentmanner.Comparedwiththenegativecontrolgroup,theapoptosisrateofMG63cellswasincreasedandthecellcyclewasarrestedattheG2/Mphase.Ontheotherside,theFasproteinexpressionwasincreasedsignificantly
7、.Conclusion:MelatonincaninhibittheproliferationofMG63ceilsbyinducingitsapoptosis.ThemechanismmaybearrestingthecellcycleattheG2/Mphaseandup-regulatingtheexpressionofFasprotein.Keywordsmelatonin;osteosarcoma;Fas;cellcycle;rea1timePCR;Westernblotting骨肉瘤是起源于間葉組織的惡性腫瘤,青春期好發(fā),報道。本研究
8、探討褪黑素對人骨肉瘤細(xì)胞MG63增殖和病因不明,對放、化療等傳統(tǒng)治療方法敏感性低,5年生凋亡的影響,并檢測其對細(xì)胞周期及凋亡相關(guān)基因Fa