• 1.西安交通大學(xué)第二醫(yī)院普外科（西安710004）;;
  • 王煒現(xiàn)在四川大學(xué)華西醫(yī)院普外科（成都610041）;;
  • 2.第四軍醫(yī)大學(xué)西京醫(yī)院消化科（西安710006）;

目的通過對半胱氨酸蛋白酶3（Caspase3）大、小亞基的重組，構(gòu)建pcDNA3.1(+)/rCaspase3真核表達(dá)質(zhì)粒,并探討rCaspase3基因誘導(dǎo)細(xì)胞凋亡的可能性，以尋求腫瘤基因治療的新途徑。方法采用分子生物學(xué)方法克隆Caspase3的大、小亞基，并在體外進(jìn)行重新排列組合，使大、小亞基原來的排序顛倒，構(gòu)建出pcDNA3.1(+)/rCaspase3真核表達(dá)質(zhì)粒； 脂質(zhì)體瞬時(shí)轉(zhuǎn)染人胰腺癌細(xì)胞PCⅡ,RTPCR檢測rCaspase3 mRNA的表達(dá)； 流式細(xì)胞技術(shù)（FCM）檢測轉(zhuǎn)染胰腺癌細(xì)胞凋亡狀況。結(jié)果Caspase3的大、小亞基被完整克隆，pcDNA3.1(+)/rCaspase3真核表達(dá)質(zhì)粒測序結(jié)果證實(shí)小亞基位于大亞基之前； RTPCR擴(kuò)增出894 bp大小片段，流式細(xì)胞檢測可見明顯的凋亡峰出現(xiàn)。結(jié)論構(gòu)建的rCaspase3其mRNA可在胰腺癌細(xì)胞中表達(dá)并自催化誘導(dǎo)細(xì)胞凋亡，可作為胰腺癌基因治療的目的基因。

引用本文： 王煒,劉志國,秦兆寅. 重組型Caspase3基因的構(gòu)建及其在胰腺癌細(xì)胞中凋亡活性的觀察. 中國普外基礎(chǔ)與臨床雜志, 2002, 9(4): 249-251. doi: 復(fù)制

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