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PubMed · 11110983

[Methods for microdissection-PCR-silver stain technique in paraffin-embedded tissue sections].

Z Huang·Q Huang·M Lai
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Abstract

OBJECTIVE: To use the microdissection-PCR-silver stain technique in the paraffin- embedded tissue sections and analyze the microsatellite instability in colorectal cancer. METHODS: By microdissection technique, the lymphocytes and mesenchymal cells, normal mucous epithelial cells, displasia, adenoma, and carcinoma cells were recovered from paraffin-embedded tissue sections. The cells were digested by protinase K; the cell lysates were used as PCR template and the PCR products were analysed by denatured polyacrylamide gel electrophoresis and silver stain. Four microsatellite loci, TGF-betaRII(A)(10), hMSH(2)(A)(26)-Bat-26, hMSH(3)(A)(8), hMSH(6)(C)(8), were analyzed. RESULTS: In all 28 specimens, hMSH(3)(A)(8) and hMSH(6)(C) 8) loci were amplified successfully. TGF-betaRII(A)10), Bat-26 had consistent amplification in 23/28, 22/28 specimens, respectively. And Bat-26 microsatellite instability was found in carcinoma cells in 5 specimens, and in adenoma cells in one of the 5 specimens. CONCLUSION: Microdissection-PCR silver stain technique can be used in paraffin-embedded tissue sections with satisfactory results. This method can be employed in analyzing microsatellite instability in colorectal carcinoma.

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Colorectal NeoplasmsDissectionHumansMicrosatellite RepeatsParaffinPolymerase Chain ReactionSilver StainingTissue EmbeddingEnglish AbstractJournal ArticleResearch Support, Non-U.S. Gov't

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Z Huang, Q Huang, M Lai. 2000. [Methods for microdissection-PCR-silver stain technique in paraffin-embedded tissue sections].. https://pubmed.ncbi.nlm.nih.gov/11110983/

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