Treatment of ras-induced cancers by the F-actin-bundling drug MKT-077.
A rhodacyanine dye called MKT-077 has shown a highly selective toxicity toward several distinct human malignant cell lines, including bladder carcinoma EJ, and has been subjected to clinical trials for cancer therapy. In the pancreatic carcinoma cell line CRL-1420, but not in normal African green monkey kidney cell line CV-1, it is selectively accumulated in mitochondria. However, both the specific oncogenes responsible for its selective toxicity toward cancer cells, and its target proteins in these cancer cells, still remain to be determined. This study was conducted using normal and ras-transformed NIH 3T3 fibroblasts to determine whether oncogenic ras mutants such as v-Ha-ras are responsible for the selective toxicity of MKT-077 and also to identify its targets, using its derivative called "compound 1" as a specific ligand. We have found that v-Ha-ras is responsible for the selective toxicity of MKT-077 in both in vitro and in vivo. Furthermore, we have identified and affinity purified at least two distinct proteins of 45 kD (p45) and 75 kD (p75), which bind MKT-077 in v-Ha-ras-transformed cells but not in parental normal cells. Microsequencing analysis has revealed that the p45 is a mixture of beta- and gamma-actin, whereas the p75 is HSC70, a constitutive member of the Hsp70 heat shock adenosine triphosphatase family, which inactivates the tumor suppressor p53. MKT-077 binds actin directly, bundles actin filaments by cross-linking, and blocks membrane ruffling. Like a few F-actin-bundling proteins such as HS1, alpha-actinin, and vinculin as well as F-actin cappers such as tensin and chaetoglobosin K (CK), the F-actin-bundling drug MKT-077 suppresses ras transformation by blocking membrane ruffling. These findings suggest that other selective F-actin-bundling/capping compounds are also potentially useful for the chemotherapy of ras-associated cancers.
Tikoo
A
A
Ludwig Institute for Cancer Research, Melbourne, Victoria, Australia.
Shakri
R
R
Connolly
L
L
Hirokawa
Y
Y
Shishido
T
T
Bowers
B
B
Ye
L H
LH
Kohama
K
K
Simpson
R J
RJ
Maruta
H
H
D016475:3T3 Cells; D000199:Actins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology / Q000648:ultrastructure; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D002462:Cell Membrane / Q000378:metabolism; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005347:Fibroblasts / Q000187:drug effects; D051379:Mice; D008819:Mice, Nude; D008854:Microscopy, Electron; D009218:Myosins / Q000378:metabolism; D009369:Neoplasms / Q000188:drug therapy*; D015689:Oncogene Protein p21(ras) / Q000378:metabolism*; D011485:Protein Binding; D011725:Pyridines / Q000494:pharmacology* / Q000627:therapeutic use*; D012513:Sarcoma, Experimental / Q000188:drug therapy; D013844:Thiazoles / Q000494:pharmacology* / Q000627:therapeutic use*; D013997:Time Factors
D000199:Actins; D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D011725:Pyridines; D013844:Thiazoles; C097880:MKT 077; D009218:Myosins; D015689:Oncogene Protein p21(ras)