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背景与目的nm23-H1基因已被证实为一种肿瘤转移抑制基因,但其作用机理尚不明了,本研究探讨nm23-H1参与肺癌Ras信号传导的机理。方法应用脂质体法将pEGFP-nm23-H1野生型和突变型质粒(WT,H118F,S120G,P96S,S44A)转染人大细胞肺癌细胞株L9981,采用免疫共沉淀与Western blot方法检测野生型和突变型nm23-H1蛋白与Ras支架蛋白KSR的关系。结果在转染了野生型和突变型质粒的五个细胞株中,鼠抗nm23-H1免疫沉淀物在86KDa处可检测到KSR的阳性条带,而各组KSR的量应用单因素方差分析比较无显著性差异(F=0.190,P=0.938)。结论本研究结果表明nm23-H1与KSR存在相互作用,但这种相互作用与nm23-H1有无突变及突变位点无关,nm23-H1可能是通过KSR参与肺癌Ras信号传导的。
Background and objective The nm23-H1 gene has been identified as a tumor metastasis suppressor gene, but its mechanism of action is unclear. This study investigated the mechanism of nm23-H1 involvement in Ras signaling in lung cancer. Methods Wild-type and mutant plasmids pEGFP-nm23-H1 (WT, H118F, S120G, P96S and S44A) were transfected into human large cell lung cancer cell line L9981 by lipofectamine. Immunoprecipitation and Western blotting were used to detect wild type Relationship between mutant nm23-H1 protein and Ras scaffold protein KSR. Results In the five cell lines transfected with wild-type and mutant plasmids, positive bands of KSR were detected at 86 kDa in mouse anti-nm23-H1 immunoprecipitation, and the amount of KSR in each group was compared by one-way ANOVA No significant difference (F = 0.190, P = 0.938). Conclusion The results of this study indicate that nm23-H1 interacts with KSR, but this interaction is not related to the presence or absence of nm23-H1 mutation and mutation. Nm23-H1 may be involved in Ras signaling in lung cancer through KSR.