Department of Pediatrics Research, Children's Cancer Hospital, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
Mol Cancer Ther. 2010 May;9(5):1111-9. doi: 10.1158/1535-7163.MCT-09-0833. Epub 2010 Apr 20.
To improve cancer outcomes, investigators are turning increasingly to small molecule medicines that disrupt vital signaling cascades, inhibit malignant growth, or induce apoptosis. One vital signaling molecule is Ras, and a key step in Ras activation is membrane anchoring of Ras through prenylation, the C-terminal addition of a lipid anchor. Small molecule inhibitors of farnesyltransferase (FTI), the enzyme most often responsible for prenylating Ras, showed clinical promise, but development of FTIs such as tipifarnib has been stalled by uncertainty about their mechanism of action, because Ras seemed unimpeded in tipifarnib-treated samples. Interpretation was further complicated by the numerous proteins that may be farnesylated, as well as availability of an alternate prenylation pathway, geranylgeranylation. Our initial observations of varied response by cancer cell lines to tipifarnib led us to evaluate the role of FTI in Ras signal alteration using various tumor models. We describe our novel counterintuitive finding that endogenous Ras activity increases in cancer cell lines with low endogenous Ras activity when farnesyltransferase is inhibited by either tipifarnib or short hairpin RNA. In response to tipifarnib, variable growth arrest and/or cell death correlated with levels of activated extracellular signal–regulated kinase (ERK) and p38 mitogenactivated protein kinase (MAPK). Sensitivity to tipifarnib treatment was shown by growth inhibition and by an increase in subdiploid cell numbers; cells with such sensitivity had increased activation of ERK and p38 MAPK. Because Ras must be prenylated to be active, our findings suggest that geranylgeranylated N-Ras or K-Ras B interacts differently with downstream effector proteins in sensitive cancer cells responding to tipifarnib, switching the balance from cell proliferation to growth inhibition [corrected].
为了改善癌症治疗效果,研究人员越来越多地转向小分子药物,这些药物可以破坏重要的信号级联反应、抑制恶性生长或诱导细胞凋亡。Ras 是一种重要的信号分子,Ras 激活的关键步骤是通过异戊烯化将 Ras 锚定在细胞膜上,即 C 端添加脂质锚。法呢基转移酶(FTI)的小分子抑制剂是最常负责异戊烯化 Ras 的酶,具有临床应用前景,但由于对其作用机制存在不确定性,法尼基转移酶抑制剂(如 tipifarnib)的开发陷入停滞,因为在 tipifarnib 处理的样本中 Ras 似乎没有受到阻碍。由于可能有许多蛋白质被法尼基化,以及存在另一种异戊烯化途径——香叶基香叶基化,因此解释变得更加复杂。我们最初观察到癌细胞系对 tipifarnib 的反应各不相同,这促使我们使用各种肿瘤模型评估 FTI 在 Ras 信号改变中的作用。我们描述了一个新颖的、违反直觉的发现,即在法尼基转移酶被 tipifarnib 或短发夹 RNA 抑制时,内源性 Ras 活性低的癌细胞系中,内源性 Ras 活性增加。对 tipifarnib 的反应与激活的细胞外信号调节激酶(ERK)和 p38 丝裂原激活的蛋白激酶(MAPK)的水平相关。通过生长抑制和亚二倍体细胞数量的增加来显示对 tipifarnib 治疗的敏感性;具有这种敏感性的细胞中 ERK 和 p38 MAPK 的激活增加。由于 Ras 必须被异戊烯化才能发挥作用,因此我们的研究结果表明,在 tipifarnib 敏感的癌细胞中,异戊烯化的 N-Ras 或 K-Ras B 与下游效应蛋白的相互作用不同,从而使细胞增殖向生长抑制转变[更正]。