目的通過對半胱氨酸蛋白酶3（Caspase3）大、小亞基的重組，構建pcDNA3.1(+)/rCaspase3真核表達質粒,并探討rCaspase3基因誘導細胞凋亡的可能性，以尋求腫瘤基因治療的新途徑。方法采用分子生物學方法克隆Caspase3的大、小亞基，并在體外進行重新排列組合，使大、小亞基原來的排序顛倒，構建出pcDNA3.1(+)/rCaspase3真核表達質粒； 脂質體瞬時轉染人胰腺癌細胞PCⅡ,RTPCR檢測rCaspase3 mRNA的表達； 流式細胞技術（FCM）檢測轉染胰腺癌細胞凋亡狀況。結果Caspase3的大、小亞基被完整克隆，pcDNA3.1(+)/rCaspase3真核表達質粒測序結果證實小亞基位于大亞基之前； RTPCR擴增出894 bp大小片段，流式細胞檢測可見明顯的凋亡峰出現。結論構建的rCaspase3其mRNA可在胰腺癌細胞中表達并自催化誘導細胞凋亡，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Caspase3基因的構建及其在胰腺癌細胞中凋亡活性的觀察. 中國普外基礎與臨床雜志, 2002, 9(4): 249-251. doi:
復制
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