Establishment of 2-dimensional electrophoresis maps of peripheral blood mononuclear cells / 中南大学学报(医学版)

OBJECTIVE@#To establish the 2-dimensional electrophoresis(2-DE) profiles of peripheral blood mononuclear cells(PBMC) in patients with hepatocellular carcinoma(HCC) and health adults.@*METHODS@#The total proteins from PBMC in patients with HCC and healthy adult were separated by immobilized pH gradient-based 2-DE. The differential expression proteins were analyzed by PDQuest analysis software.@*RESULTS@#The well-resolved, reproducible 2-DE patterns of PBMC in patients with HCC and healthy adults were obtained. For HCC, the average spots of 2-DE maps were 1 206 +/- 48, and the average matching rate was 90.8%. For normal adults, the average spots were 1 123 +/- 37, and the average matching rate was 92.6%.@*CONCLUSION@#The well-resolved, reproducible 2-DE patterns of PBMC in patients with HCC and healthy adults are established. These proteomic analysis methods are useful to screen the potential biomarkers in the early diagnosis, treatment and prognosis monitor in patients with malignant tumor.

Mutation analysis of the checkpoint kinase 2 gene in colorectal cancer cell lines / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Checkpoint kinase 2 (CHK2) is a DNA damage-activated protein kinase which is involved in cell cycle checkpoint control. CHK2 gene could be a candidate gene for colorectal cancer susceptibility. But there are few systematic reports on mutation of CHK2 in colorectal cancer.</p><p><b>METHODS</b>The mutations of all 14 exons of CHK2 in 56 colorectal cancer cell lines were screened systematically, using denaturing high-performance liquid chromatography (DHPLC) to screen the mismatches of the CHK2 exons amplified products, and then the suspected mutant cell lines were scanned by nucleotide sequence analysis.</p><p><b>RESULTS</b>VACO400 in CHK2 exon 1a was suspected to have mutation by DHPLC and confirmed by sequence, but this was nonsense mutation. C106, CX-1, HT-29, SK01, SW480, SW620 and VACO400 in CHK2 exon 1b were confirmed to have the same nonsense mutation in 11609 A > G. DLD-1 and HCT-15 in CHK2 exon 2 were confirmed to have missense mutation R145W, which was heterozygous C > T missense mutation at nucleotide 433, leading to an Arg > Trp substitution within the FHA domain.</p><p><b>CONCLUSIONS</b>The CHK2 mutation in colorectal cancer is a low frequency event. There are just 10 cell lines to have sequence variations in all the 14 exons in 56 colorectal cancer cell lines and only DLD-1/HCT-15 had heterozygous missense mutation. These findings may give useful information of susceptibility of colorectal cancer as single nucleotide polymorphysim.</p>