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重组K-Ras突变基因在胰腺癌Bxpc-3细胞株中的表达及其对化疗药物药效的影响

, PP. 1152-1158

Keywords: 肿瘤化疗,抗肿瘤药物,K-Ras突变,质粒,胰腺癌

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Abstract:

K-Ras是Ras基因家族中的一员,在细胞生存、增殖和分化过程中发挥重要作用.当Ras基因发生突变时,其表达的Ras蛋白可能被过度激活,从而导致肿瘤的产生和发展.胰腺癌中K-Ras的突变频率较高,但其对胰腺癌化疗药物药效的影响尚不明确.本文以pAcGFP1-C3质粒为载体构建了含有5种不同基因型K-Ras序列的重组质粒,分别为野生型和G12V,G12R,G12D,G13D4种突变型.将上述重组质粒转入人胰腺癌Bxpc-3细胞株并筛得稳转株.结果显示,转染野生型和突变型K-Ras的细胞株具有相似的生长速度,且细胞中K-Ras-GFP融合蛋白的表达量也一致.药效学研究显示,转染突变型K-Ras基因的细胞对吉非替尼、5-氟尿嘧啶、多西他赛和吉西他滨的药物敏感性降低,而对厄罗替尼和顺铂的药效则无显著影响,这可能与转染突变型K-Ras基因的细胞中K-Ras下游信号p-Akt和p-Erk的水平升高有关.此外,转染不同突变型K-Ras导致的药效及p-Akt,p-Erk的表达变化也不同.体外研究结果表明,K-Ras突变细胞对多种化疗药物的药效敏感性降低,同时不同突变型细胞之间药效也有差异,该结果有待进一步深入研究.

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