K-Ras突变对表皮生长因子受体抑制剂敏感细胞株的影响

目的 观察K-Ras突变对于携带表皮生长因子受体(EGFR)突变细胞的EGFR抑制剂敏感性的影响.方法 构建K-Ras突变真核表达质粒,采用脂质体转染技术转染肺癌细胞HCC827(EGFR突变,K-Ras野生)和H292(EGFR、K-Ras均野生),噻唑蓝(MTT)比色法测定转染K-Pas突变质粒和空白质粒后各细胞对EGFR抑制剂的半数抑制浓度(IC_(50)).结果 真核表达质粒构建成功.细胞HCC827未转染K-ras突变质粒对吉非替尼(Iressa)的IC_(50)为0.007,转染后对Iressa的IC_(50)为12.3,差异有统计学意义(P<0.01).细胞H292未转染K-ras突变质粒对Iressa的IC_(50)为0.04,转染后对Iressa的IC_(50)为12.0,差异有统计学意义(P<0.01).细胞A549(K-Ras突变)对Iressa的IC_(50)为12.8,与转染K-ras突变质粒的细胞HCC827及H292比较,差异无统计学意义(P>0.05).结论 野生型或突变型EGFR出现K-Ras突变均可引起吉非替尼耐药,其程度与K-Ras突变的细胞株相当.

The influence of K-Ras mutation on epidermal growth factor receptor inhibitor sensitive cell lines

Objective By comparing the sensitivity of gefitinib in different cell lines affected by K-Ras mutation,to investigate the change of epidermal growth factor receptor(EGFR)inhibitors sensitivity on EGFR mutation cells with K-Ras mutation.Methods The eukaryotic expression plasmid pcDNA3.1 (-)K-Ras(+)was constyructed,transfected into lung cancer cells HCC827(EGFR mutation,K-Ras Wild-type)and H292(EGFR Wild-type,K-Ras Wild-type)by liposome respectively.MTT was used to measured the semitivity and median lethal concentration IC_(50) of EGFR inhibitors gefitinib after K-Ras or blank plasmid was induced to ceils.Results The sequencing results confirmed the success of eukaryotic expression plasmid.IC_(50) H of gefitinib for HCC827(K-Ras mutation)and HCC827(K-Ras Wild-type)was 12.3,and 0.007(P<0.01)respectively;that for H292(K-Ras mutation)and H292(K-Ras Wild-type)was 12.0,and 0.04(P<0.01)respectively;and that for A549(K-Ras mutation)12.8(P>0.05).Conclusion Whether with EGFR mutations or not,the sensitivity to gefitinib waft significantly decreased with K-Ras mutant transfection,and the extent of resistance Was close to cells carrying K-Ras mutation.