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1、·基礎論著·二硫化二砷誘導T淋巴細胞白血病細胞株凋亡的機制研究王玲【摘要】目的探討二硫化二砷誘導T淋巴細胞白血病細胞株凋亡的分子機制。方法將不同濃度的二硫化二砷與T淋巴細胞白血病細胞株共培養(yǎng)。采用CCK-8方法測定細胞活性;AnnexinV-PI方法測定細胞凋亡率,采用實時定量PCR在基因水平測定凋亡相關因子Bc1.2和Bax的表達。結果隨著二硫化二砷藥物濃度的增高,細胞活性減低,隨著藥物作用時間的延長,細胞活性逐漸減弱。其差異均有統(tǒng)計學意義。細胞凋亡結果顯示:二硫化二砷藥物濃度越高,細胞凋亡率越高,隨著藥物作用時間的延長,細胞凋亡率也越高。其差異均有統(tǒng)計學意義。實時熒光定量結果顯示:
2、與對照組比較,10grnol/L二硫化二砷與CEM細胞共孵育48h,Bax/Bc1.2的比例升高(P<0.05)。結論二硫化二砷以時間和劑量依賴的方式抑制CEM細胞生長和誘導凋亡,誘導凋亡的機制與Bax/Bcl-2的比率增加有關?!娟P鍵詞】白血病,T細胞;細胞凋亡;bc1.2相關x蛋白質;基因,bc1.2;二硫化二砷ThemechanismofarsenicsulfideinducedapoptosisofTlymphocyteleukemiacellsWANGLing.DepartmentofHematology,TaiancfCentralHospital,Taian271000,
3、ChinaEmaibwang177@163.com【Abstract】objectiveToinvestigatethemechanismofarsenicsulfideinducedapoptosisofTlymphocyteleukemiacelllineCEM.MethodsCEMceilsweretreatedwithvariousconcentrationsofArsenicsulfidefordiferenttimeperiods.CellviabilitywasdetectedbytheCCK-8assay.cellapoptosiswasexaminedbyflowcy
4、tometry.ThemRNAlevelofBaxandBcl一2wasdetectedbyRT-PCR.ResuRsWefoundthattheTlymphocyteleukemiacellsviabilitywassignificantlydecreasedfollowingtreatment、mArsenicsulfidefor24h.48h.Alongwithincreasingarsenicsulfideconcentration,thecellsviabilitywasnotablyreducedwhencomparedwiththecontrolgroup,andther
5、esultswerestatisticallysignificant.Theapoptoticratesofcellsweresignificantlyincreasedat24hand48hwithincreasingarsenicsulfideconcentration,andwere~atisticaHysignificant.ThequantRativereal--timePCRresultsshowedthattheexpressionlevelsofBax/Bcl-2ratiowereup-—regulatedinArsenicsulfide-treatedcells.Co
6、nclusionsArsenicsulfideinhibitedproliferationandinducedapoptosisofTlymphocyteleukemiacellsinadose-andtime—dependentmanner.ThemechanismofarsenicsulfideinducedapoptosisofTlymphocyteleukemiacellswasrelatedtoBax/Bcl一2upregulatiOI1.【Keywords1Leukemia,T—cell;Apoptosis;bcl一2一associatedXprotein;Genes,bc
7、l一2;Arsenicsul6de雄黃的主要成分是二硫化二砷或四硫化四砷,由材料與方法于雄黃的抗腫瘤作用使其成為抗腫瘤的熱點,尤其是對惡性血液病的治療其效果顯著。本實驗采用高純度1.細胞株與藥物:人T淋巴細胞白血病細胞株的二硫化二砷(arsenicsulfide),體外觀察二硫化二CEM培養(yǎng)于l0%的胎牛血清(fetalbovineserum,F(xiàn)BS)砷對T淋巴細胞白血病細胞株CEM的凋亡作用,并探的RPMI培養(yǎng)基中,在37℃、5%CO2、飽和