Di Paolo A, Danesi R, Caputo S, Macchia M, Lastella M, Boggi U, Mosca F, Marchetti A, Del Tacca M
Division of Pharmacology and Chemotherapy, University of Pisa, Italy.
Br J Cancer. 2001 Jun 1;84(11):1535-43. doi: 10.1054/bjoc.2001.1820.
Proteins belonging to the ras superfamily are involved in cell proliferation of normal and neoplastic tissues. To be biologically active, they require post-translational isoprenylation by farnesyl-transferase and geranylgeranyl-transferase. Enzyme inhibition by drugs may thus represent a promising approach to the treatment of cancer. Therefore, the combined effect of BAL9611, a novel inhibitor of geranylgeranylation, and manumycin, a farnesyl-transferase inhibitor, was evaluated on the SW620 human colon cancer cell line which harbours a mutated K-ras gene. BAL9611 and manumycin dose-dependently inhibited SW620 cell growth with 50% inhibitory concentration (IC(50)) of 0.47 +/- 0.03 and 5.24 +/- 1.41 microM (mean +/- SE), respectively. The isobologram analysis performed at the IC(50)level revealed that the combined treatment was highly synergistic with respect to cell growth inhibition. BAL9611 and manumycin were able to inhibit the geranylgeranylation of p21rhoA and farnesylation of p21ras; both drugs inhibited p42ERK2/MAPK phosphorylation, but their combination was more effective than either drug alone. Moreover, the enhanced inhibition of cell growth in vitro by the BAL9611-manumycin combination was also observed in vivo in CD nu/nu female mice xenografted with SW620 tumours. Finally, both drugs were able to induce cell death by apoptosis in vitro and in vivo, as demonstrated by perinuclear chromatin condensation, cytoplasm budding and nuclear fragmentation, and interoligonucleosomal DNA digestion. In conclusion, the inhibition of protein farnesylation enhances the chemotherapeutic effect of BAL9611 in vitro and in vivo in a synergistic fashion, as a result of the impairment of post-translational isoprenylation of proteins and phosphorylation of p42ERK2/MAPK, whose activation is associated with post-translational geranylgeranylation and farnesylation of p21rhoA and p21ras.
属于ras超家族的蛋白质参与正常组织和肿瘤组织的细胞增殖。为了具有生物活性,它们需要通过法尼基转移酶和香叶基香叶基转移酶进行翻译后异戊二烯化修饰。因此,药物对酶的抑制作用可能是一种很有前景的癌症治疗方法。因此,研究了新型香叶基香叶基化抑制剂BAL9611和法尼基转移酶抑制剂马尼霉素对携带K-ras基因突变的SW620人结肠癌细胞系的联合作用。BAL9611和马尼霉素均剂量依赖性地抑制SW620细胞生长,其50%抑制浓度(IC50)分别为0.47±0.03和5.24±1.41微摩尔/升(平均值±标准误)。在IC50水平进行的等效线图分析显示,联合治疗在抑制细胞生长方面具有高度协同作用。BAL9611和马尼霉素能够抑制p21rhoA的香叶基香叶基化修饰和p21ras的法尼基化修饰;两种药物均抑制p42ERK2/MAPK磷酸化,但它们的联合作用比单独使用任何一种药物都更有效。此外,在接种SW620肿瘤的CD nu/nu雌性小鼠体内也观察到BAL9611-马尼霉素联合用药在体外对细胞生长的抑制作用增强。最后,两种药物在体外和体内均能通过凋亡诱导细胞死亡,表现为核周染色质浓缩、细胞质出芽和核碎裂以及寡核苷酸间DNA消化。总之,由于蛋白质翻译后异戊二烯化修饰和p42ERK2/MAPK磷酸化受损,蛋白质法尼基化修饰的抑制以协同方式增强了BAL9611在体外和体内的化疗效果,而p42ERK2/MAPK的激活与p21rhoA和p21ras的翻译后香叶基香叶基化修饰和法尼基化修饰相关。