A. Umar et al., DEFECTIVE MISMATCH REPAIR IN EXTRACTS OF COLORECTAL AND ENDOMETRIAL CANCER CELL-LINES EXHIBITING MICROSATELLITE INSTABILITY, The Journal of biological chemistry, 269(20), 1994, pp. 14367-14370
A replication error (RER(+)) phenotype, characterized by somatic insta
bility in simple repeated sequences, is associated with several types
of cancer. To determine if a defect in DNA replication fidelity or rep
air of replication errors might explain this instability, we compared
both processes in cell-free extracts from RER(+) endometrial and color
ectal cancer cell lines to RER(-) cell lines. SV40 origin-dependent re
plication of a microsatellite sequence is highly accurate in cell extr
acts regardless of their RER phenotype. However, extracts from RER(+)
cell lines are defective in mismatch repair, while extracts of RER(-)
cell lines are not. Lack of repair was observed when the signal (a nic
k) for strand specific repair was either 3' or 5' to the mispair. One
colorectal cancer cell line contained deletions in both alleles of the
putative mismatch repair gene hMSH2, and one endometrial cancer cell
line contained a 4-base pair duplication in one hMSH2 allele. No hMSH2
mutation was detected in the other allele or in the other five RER(+)
cell lines. Repair was readily detected when each of the defective ex
tracts was mixed with a repair-proficient extract, demonstrating that
no trans-acting inhibitor is present. Attempts to complement the repai
r deficiencies by mixing two different defective extracts identified t
hree combinations that restored repair. The data suggest that: (i) def
ective repair is associated with colorectal and endometrial cancer and
, by extrapolation, with other types of cancer; (ii) mutations in the
hMSH2 gene, and possibly other genes, result in defective mismatch rep
air; (iii) the defect(s) in these lines likely involves pre-incision e
vents or the excision step, but not the incision, polymerization, or l
igation steps; and (iv) at least four functional complementation group
s for mismatch repair may be involved in human cancer.