Thalidomide: lack of mutagenic activity across phyla and genetic endpoints.
The human and rabbit teratogen thalidomide has been tested for mutagenicity in a wide range of assays, ranging from bacterial gene mutation assays conducted in vitro to in vivo cytogenetic assays conducted using rabbits, and including a variety of human-derived tissues. Thalidomide was not mutagenic to 6 strains of Salmonella when tested both in the presence and absence of Aroclor-induced rat liver S9 mix. This inactivity was confirmed in strains TA98 and TA100 using a 1-h pre-incubation assay protocol with the same S9 mix (10% S9), and additionally, in strain TA98 using 3 concentrations of S9 (4%, 10% and 30% S9 in S9 mix). Thalidomide was not clastogenic either to cultured human lymphocytes (whole blood cultures, minus S9 mix) or to Chinese hamster ovary (CHO) cells treated in vitro. Further, no cytotoxicity was observed in purified human lymphocytes when exposed to thalidomide up to the limit of its solubility in the medium in the presence and absence of liver S9 from Aroclor-induced pregnant rabbit. The CHO assays were conducted without metabolic activation and in the presence of a variety of sources of auxiliary metabolic activation (PB/beta NP-induced rat liver S9 mix, pooled male and female human liver S9 mix, uninduced and Aroclor-induced pregnant rabbit liver S9 mix and foetal rabbit S9 mix). Thalidomide did not induce micronuclei in isolated human lymphocytes (minus S9 mix) and it was non-mutagenic to mouse lymphoma L5178Y TK+/- cells when tested to the limits of its solubility in the culture medium (+/- S9 mix). No indication of recombinogenic or clastogenic activity was observed for thalidomide when tested in Drosophila. In addition, it failed to induce chromosome aberrations in grasshopper neuroblasts when tested in the presence and absence of Aroclor-induced rat liver S9 mix. Some unusual chromosome morphologies were observed in the grasshopper cytogenetic preparations indicating a potential of thalidomide to interact with chromosomal proteins. However, this potential was not evident in the human lymphocyte micronucleus assay, and thalidomide was apparently not reactive to the proteins of the mouse skin, as it gave negative results in a mouse local lymph node assay for skin sensitizing agents. Thalidomide was inactive in bone marrow micronucleus assays conducted using males and females from two strains of mice, and female New Zealand white rabbits. It is concluded that thalidomide is neither a mutagen nor an aneugen. This conclusion is discussed within the context of the results of earlier mutagenicity studies, the recent claim that thalidomide may be a heritable germ cell mutagen to humans, and the current interest in thalidomide for the treatment of immune system-related diseases.
Ashby
J
J
Zeneca CTL, Macclesfield, Cheshire, UK.
Tinwell
H
H
Callander
R D
RD
Kimber
I
I
Clay
P
P
Galloway
S M
SM
Hill
R B
RB
Greenwood
S K
SK
Gaulden
M E
ME
Ferguson
M J
MJ
Vogel
E
E
Nivard
M
M
Parry
J M
JM
Williamson
J
J
D000014:Abnormalities, Drug-Induced / Q000209:etiology; D000818:Animals; D001711:Biotransformation; D016466:CHO Cells / Q000187:drug effects; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D006224:Cricetinae; D004331:Drosophila melanogaster / Q000187:drug effects; D005260:Female; D006110:Grasshoppers; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D015162:Micronucleus Tests; D008862:Microsomes, Liver / Q000378:metabolism; D009152:Mutagenicity Tests; D009474:Neurons / Q000187:drug effects; D011247:Pregnancy; D011817:Rabbits; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects; D013045:Species Specificity; D013234:Stem Cells / Q000187:drug effects; D013723:Teratogens / Q000633:toxicity; D013792:Thalidomide / Q000633:toxicity*
D013723:Teratogens; D013792:Thalidomide