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本帖最后由 细胞海洋 于 2012-3-26 13:34 编辑
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- d% v+ L# F- o+ {' a; tUse of Comet-FISH in the study of DNA damage and repair:$ t) `+ K1 }. J7 O/ }& y! r
The Comet-FISH technique is a useful tool to detect overall and region-specific DNA damage and repair in individual cells. It combines two
: J* K; I5 P4 A* t4 _/ o' l& kwell-established methods, the Comet assay (single cell gel electrophoresis) and the technique of fluorescence in situ hybridization (FISH). Whereas
- K0 @% ^4 A! nthe Comet assay allows separating fragmented from non-fragmented DNA, FISH helps to detect specifically labelled DNA sequences of interest,
4 k) z! {- B/ i) I E. r7 Bincluding whole chromosomes. Thus the combination of both techniques has been applied in particular for detection of site-specific breaks in DNA; H+ i# e6 _' v/ \7 x
regions which are relevant for development of different diseases. This paper reviews the relevant literature and presents three examples on how4 ^; A# k7 M Q" I9 M" o- {
Comet-FISH was used for studying the induction of DNA damage by genotoxic compounds related to oxidative stress in colon cancer-relevant
4 ~* s. ]! ^9 }( qgenes (TP53, APC, KRAS) of a colon adenoma cell line. The accumulated evidence on relative sensitivity of these genes in comparison to global
. A2 c! A: g' u/ L, z- ~$ y' }! Odamage allows a more definite conclusion on the possible contribution of the genotoxic factors during colorectal carcinogenesis. Telomere fragility
* r; \( W& G. M6 c2 u9 U6 S. J- uwas compared in different cell lines treated with cytostatic agents, and revealed new patterns of biological activities through the drugs and different$ z3 P9 K. d0 `
sensitivities of the cell lines that were found to be associated with their tumour origin. A third example relates to measuring repair of specific gene. s$ ~' l( S& I
regions using Comet-FISH, a method that can be developed to biomarker application. Taken together, available data suggests that Comet-FISH
' c: j' D( y3 T% t+ Zhelps to get further insights into sensitivity of specific DNA regions and consequently in mechanisms of carcinogenesis. Although the nature of the
# H# r& ]( @1 \- h9 q" e4 bmeasured Comet-FISH endpoint precludes us from stating basically that damage and repair are occurring within the specific gene, it is at least
9 N, g' b9 \% v* ipossible to evaluate whether the damage and repair are occurring within the vicinity of the gene of interest.) Q; G$ x$ X- ~8 ]& y# j+ P3 N1 ?
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发表于 2012-3-26 10:54
