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含hTERT启动子、PE38KDEL片段质粒构建及其对人胰腺癌细胞凋亡的影响 被引量:2

Construction of recombinant plasmid containing hTERT promoter and PE38KDEL gene and its anti-apoptosis effect on human pancreatic cancer cell MiaPaCa2
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摘要 目的构建含hTERT启动子和PE38KDEL片段的重组表达质粒,并观察其对人胰腺癌MiaPaCa2细胞凋亡的影响。方法采用PCR法构建含hTERT启动子和PE38KDEL片段的重组表达质粒,并用双酶切法鉴定;将MiaPaCa2细胞分为5组,A组为空白对照组,仅加入含10%FBS的DMEM;B、C、D、E组分别转染pIRES2-EGFP、phTERT-IRES2-EGFP、pPE38KDEL-IRES2-EGFP、phTERTp-PE38KEDL-IRES2-EGFP,采用TUNEL法检测MiaPaCa2细胞的凋亡指数。结果重组质粒的PCR产物及酶切后片段的测序结果与GeneBank报道序列一致。A组转染24、48 h时MiaPaCa2细胞凋亡率分别为17.12%±1.66%、24.86%±2.80%,B组分别为18.14%±1.61%、26.47%±2.71%,C组为19.08%±3.84%、27.42%±7.26%,D组为29.46%±2.86%、38.12%±3.19%,E组为72.16%±4.79%、86.40%±7.71%。B^E组与A组比较,P均<0.05;E组与D组比较,P<0.05。结论成功构建了含hTERT启动子和PE38KDEL片段的重组表达质粒,该质粒可促进人胰腺癌MiaPaCa2细胞的凋亡。 Objective To construct a recombinant plasmid containing hTERT promoter and PE38KDEL gene and observe its anti-apoptosis effects on human pancreatic cancer cell MiaPaCa2.Methods The recombinant plasmid contain hTERTpromoter and PE38KDEL gene(phTERTp-PE38KDEL-IRES2-EGFP)was constructed by PCR and double digestion methods.phTERTp-PE38KDEL-IRES2-EGFP was transfected to MiaPaCa2 cell and apoptotic index of MiaPaCa2 cell were detected by TUNEL.Results The recombinant plasmid phTERTp-PE38KDEL-IRES2-EGFP was highly expressed in human pancreatic cancer cell line MiaPaCa2 of telomerase positive,and it could selective killed MiaPaCa2 in vitro.Conclusion The recombinant plasmid phTERTp-PE38KDEL-IRES2-EGFP is constructed successfully,and it can kill human pancreatic cancer cell line MiaPaCa2 specifically.
出处 《山东医药》 CAS 北大核心 2010年第34期4-6,共3页 Shandong Medical Journal
基金 教育部博士点基金资助项目(20060062008) 天津市自然科学基金自助项目(07JCYBJC10300)
关键词 胰腺癌 人端粒酶反转录酶 启动子 铜绿假单胞菌外毒素 绿色荧光蛋白 基因疗法 pancreatic cancer green fluorescent protein human telomerase reverse transcriptase promoter pesudomonas aeruginosa exotoxin gene therapy
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参考文献8

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同被引文献13

  • 1孔北华,宋悦,马道新,曲迅,江森.人端粒酶逆转录酶启动子调控下胞嘧啶脱氨酶胸苷激酶融合基因治疗卵巢癌的体外研究[J].中华妇产科杂志,2004,39(6):390-395. 被引量:11
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  • 7Moriai M,Tsuji N,Kobayashi D,et al.Down-regulation of hTERT expression plays an important role in 15-deoxy-12,14-prostaglandin J2-induced apoptosis in cancer cells[J].J Oncol,2009,34(5):1363-1372.
  • 8Glinka E M,Andryushchenko A S,Sapozhnikov A M,et al.Construction of the plasmid for expression of ETA-EGFP fusion protein under control of the cytomegalovirus promoter and its effects in HeLa cells[J].Plasmid,2009,62(2):119-127.
  • 9宋悦,沈铿,于靖蓉.人端粒酶逆转录酶启动子调控下活性半胱氨酸蛋白酶-3治疗人卵巢癌[J].中华医学杂志,2007,87(41):2919-2924. 被引量:2
  • 10刘杰,阚丹,刘立思.IKK/IκB/NF-κB信号通路阻断及临床应用[J].实用医学杂志,2008,24(23):4147-4149. 被引量:17

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