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


CONCLUSIONS
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.
Huang
Z
Z
Department of Pathology, Medical College of Zhejiang University, Hangzou, Zhejiang, 310031 P.R.China. huangzhd@mail.hz.zj.cn
Huang
Q
Q
Lai
M
M
D015179:Colorectal Neoplasms / Q000235:genetics*; D004210:Dissection; D006801:Humans; D018895:Microsatellite Repeats; D010232:Paraffin; D016133:Polymerase Chain Reaction; D016622:Silver Staining; D016610:Tissue Embedding
D010232:Paraffin