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1、熊果酸通過影響B(tài)ax和Bcl作者: 張貴平,黎銀燕,呂嘉春,區(qū)彗堅(jiān)【關(guān)鍵詞】細(xì)胞凋亡UrsolicacidinducesapoptosisthrougheffectsonBaxandBcl2expressioninhumanMCF7cells 【Abstract】AIM:Tostudytheeffectsofursolicacid(UA)onhumanMCF7cellapoptosisandthemechanismprobablyinvolved.METHODS:MCF7cellsol/L)for24h,apoptoticcellsentednuc
2、lEiicroscope.AfterMCF7cellstreatedol/L)for24h,subG1peakandcellcycleol/LofUAfor24h,theprotEInexpressionofcytochromec,Bcl2andBaxmunofluorescencecellstaining(SABCCy3)andthegreylevelsofimmunofluorescenceoncytochromecandBcl2andBaxent,apoptoticcellsincreaseddosedependentlyandthemorph
3、ologicalchangesofMCF7cellsdisplayedmanyhallmarkfeaturesofapoptosis,includingchromatinaggregationandfragmentednuclei.SubG1ratesin0,20and50μmol/LofUAdetectedbyFCML/LDMSO,50μmol/LofUAsignificantlyincreasedthegreylevelsofBax(59.3±7.8,213.6±7.4,P<0.01),cytochromec(88.2±6.9,188.1±
4、15.4,P<0.01)andthegreyratioofBax/Bcl2(1.0±0.1,2.4±0.4,P<0.01)andincreasednoticeablythegreylevelofBcl2(58.1±6.1,92.1±12.4,P<0.01).Cytochromecinducedby50μmol/LofUAreleasedintocytoplasm.CONCLUSION:ApoptosisinducedecdependentapoptoticpathaybeapotentialChinesemedicinalponen
5、tforbreastneoplasm. 【Keyec 【摘要】目的:探討中藥成分熊果酸(ursolicacid,UA)誘導(dǎo)MCF7人乳腺癌細(xì)胞的凋亡作用,并通過分析UA作用后的MCF7細(xì)胞Bax,Bcl2,cytochromec蛋白表達(dá)的變化,探討其誘導(dǎo)MCF7細(xì)胞凋亡的分子機(jī)制.方法:采用細(xì)胞培養(yǎng)技術(shù),用0,10,30和50μmol/L的UA處理MCF7細(xì)胞24h,用Ho染細(xì)胞,在熒光顯微鏡下觀察凋亡細(xì)胞的形態(tài)變化.用0,20和50μmol/L的UA處理MCF7細(xì)胞24h,用PI染細(xì)胞,在流式細(xì)胞儀上測定細(xì)胞周期的變化.用0和50μmol/L的
6、UA處理MCF7細(xì)胞24h,采用熒光免疫細(xì)胞化學(xué)SABCCy3法檢測Bax,Bcl2,cytochromec蛋白表達(dá),在熒光顯微鏡下拍照,用QCF7細(xì)胞24h,隨著UA濃度增加,在熒光顯微鏡下出現(xiàn)凋亡細(xì)胞增多,細(xì)胞呈現(xiàn)典型的凋亡形態(tài)學(xué)特征,核質(zhì)濃集和凋亡小體.用0,20和50μmol/L的UA處理MCF7細(xì)胞24h,流式細(xì)胞儀檢測亞G1峰比例分別為0.05,0.22,0.43與對照組5mL/LDMSO比較,50μmol/L的UA使MCF7細(xì)胞中的Bax灰度(59.3±7.8,213.6±7.4,P<0.01)和cytochromec灰度(88.
7、2±6.9,188.1±15.4,P<0.01)增加,Bax/Bcl2灰度比值升高(1.0±0.1,2.4±0.4,P<0.01),Bcl2灰度(58.1±6.1,92.1±12.4,P<0.01)稍有增加.UA促進(jìn)線粒體放釋cytochromec進(jìn)入胞質(zhì).結(jié)論:UA誘導(dǎo)MCF7細(xì)胞凋亡,其機(jī)制涉及到Bax/Bcl2比值升高引起線粒體釋放cytochromec所依賴的凋亡調(diào)節(jié)信號通路.表明治療乳腺癌,UA可能是有效的中藥成分. 【關(guān)鍵詞】熊果酸;MCF7細(xì)胞;細(xì)胞凋亡;基因,Bax;基因,Bcl2;細(xì)胞色素C類 0 引言 熊果
8、酸(ursolicacid,UA),屬三萜類化合物,自然資源豐富.許多研究表明,UA具有誘導(dǎo)多種腫瘤細(xì)胞凋亡