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《二硫化二砷誘導(dǎo)T淋巴細(xì)胞白血病細(xì)胞株凋亡的機(jī)制研究-論文.pdf》由會員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在應(yīng)用文檔-天天文庫。
1、·基礎(chǔ)論著·二硫化二砷誘導(dǎo)T淋巴細(xì)胞白血病細(xì)胞株凋亡的機(jī)制研究王玲【摘要】目的探討二硫化二砷誘導(dǎo)T淋巴細(xì)胞白血病細(xì)胞株凋亡的分子機(jī)制。方法將不同濃度的二硫化二砷與T淋巴細(xì)胞白血病細(xì)胞株共培養(yǎng)。采用CCK-8方法測定細(xì)胞活性；AnnexinV-PI方法測定細(xì)胞凋亡率，采用實(shí)時定量PCR在基因水平測定凋亡相關(guān)因子Bc1．2和Bax的表達(dá)。結(jié)果隨著二硫化二砷藥物濃度的增高，細(xì)胞活性減低，隨著藥物作用時間的延長，細(xì)胞活性逐漸減弱。其差異均有統(tǒng)計(jì)學(xué)意義。細(xì)胞凋亡結(jié)果顯示：二硫化二砷藥物濃度越高，細(xì)胞凋亡率越高，隨著藥物作用時間的延長，細(xì)胞凋亡率也越高。其差異均有統(tǒng)計(jì)學(xué)意義。實(shí)時熒光定量結(jié)果顯示：
2、與對照組比較，10grnol／L二硫化二砷與CEM細(xì)胞共孵育48h，Bax／Bc1．2的比例升高(P<0．05)。結(jié)論二硫化二砷以時間和劑量依賴的方式抑制CEM細(xì)胞生長和誘導(dǎo)凋亡，誘導(dǎo)凋亡的機(jī)制與Bax／Bcl-2的比率增加有關(guān)。【關(guān)鍵詞】白血病，T細(xì)胞；細(xì)胞凋亡；bc1．2相關(guān)x蛋白質(zhì)；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(diǎn)，尤其是對惡性血液病的治療其效果顯著。本實(shí)驗(yàn)采用高純度1．細(xì)胞株與藥物：人T淋巴細(xì)胞白血病細(xì)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(xì)胞白血病細(xì)胞株CEM的凋亡作用，并探的RPMI培養(yǎng)基中，在37℃、5％CO2、飽和
