Identification of a Second MutL DNA Mismatch Repair Complex (hPMS1 and hMLH1) in Human Epithelial Cells

Leung, Wai K.; Kim, Jae J.; Wu, Ling; Sepulveda, Jorge L.; Sepulveda, Antonia R.

doi:10.1074/jbc.m908768199

Cited by 59 publications

(51 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Protein was extracted as described previously (Leung et al, 2000a). Gastric tissues and cell lines were homogenized in Tris-HCl (pH 7.4) buffer containing 0.5% Triton X-100 and protease inhibitor cocktail (Roche, Indiapolis, IN, USA).…”

Section: Western Blotmentioning

confidence: 99%

Increased expression of survivin in gastric cancer patients and in first degree relatives

Leung

Ebert

et al. 2002

Br J Cancer

View full text Add to dashboard Cite

Survivin was recently described as an apoptosis inhibitor. Its pathogenic role in gastric cancer is largely unknown. Expression of survivin in gastric cancer and non-cancer first-degree relatives, and its association with apoptosis and cyclo-oxygenase-2 expression was investigated. Fifty gastric cancer, 30 non-cancer first-degree relatives, 20 normal controls and five gastric cancer cell lines were studied. Survivin and cyclo-oxygenase-2 were evaluated by reverse transcriptase-polymerase chain reaction, immunohistochemistry and Western blot. Survivin expression was absent from normal gastric mucosa. All five cancer cell lines and 34 out of 50 (68%) human gastric cancer tissues expressed survivin mRNA. Survivin expression was less frequent (22%; P50.001) in adjacent non-tumour gastric tissues. Immunohistochemistry and Western blot obtained similar findings. Gastric cancers with survivin expression displayed significantly reduced apoptosis (P=0.02), and associated with cyclo-oxygenase-2 overexpression at both mRNA (P=0.001) and protein levels (P=0.041). Moreover, survivin mRNA was detected in the gastric mucosa of eight (27%) non-cancer relatives. Expression in non-cancer patients showed positive correlation with H. pylori infection (P=0.004). This demonstrates the frequent expression of survivin in gastric cancer and in first-degree relatives. Coexpression of survivin and cyclo-oxygenase-2 may suggest multiple pathways contributing to the inhibition of apoptosis in gastric cancer.

show abstract

Section: Western Blotmentioning

confidence: 99%

Increased expression of survivin in gastric cancer patients and in first degree relatives

Leung

Ebert

et al. 2002

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…The cell lines SNU638, SNU668 and SNU719 were previously reported. 3,27 The cell lines MKN1, MKN7, MKN28, MKN74, GTL16 and TMK1, were kindly given by Dr Richard Hamelin and were previously reported. 28,29 The cells were grown in RPMI-1640 medium containing 10% fetal bovine serum, penicillin and streptomycin (GIBCO BRL, Gaithersburgh, MD, USA), except KATOIII cells that were grown in RPMI-1640 containing 20% fetal bovine serum and penicillin and streptomycin.…”

Section: Gastric Cancer Cell Linesmentioning

confidence: 99%

“…Genomic DNA was extracted from 10 separate cell clones per marker examined, as previously described. 3 The microsatellite markers included the BAT25 and BAT26 mononucleotide markers and the D2S123, D17S250, D13S170 dinucleotide repeat markers. 3,15 The oligonucleotide primers were purchased from Research Genetics, Inc. Huntsville, AL, USA.…”

Section: Msi Analysismentioning

confidence: 99%

“…3 The microsatellite markers included the BAT25 and BAT26 mononucleotide markers and the D2S123, D17S250, D13S170 dinucleotide repeat markers. 3,15 The oligonucleotide primers were purchased from Research Genetics, Inc. Huntsville, AL, USA. One of the primers was end-labeled with g 32 P-ATP using T4 polynucleotide kinase (Promega Corp., Madison, WI, USA).…”

Section: Msi Analysismentioning

confidence: 99%

“…Efficient DNA mismatch repair requires the combined function of MutL (hMLH1-hPMS2 and hMLH1-hMLH3) and MutS (hMSH2-hMSH6 and hMSH2-hMSH3) heterodimers. [1][2][3][4][5] In MMR-deficient cells, insertion or deletion of nucleotide bases at microsatellite sequences causing microsatellite instability (MSI)-type mutations occur at high frequency. 1 Inactivation of genes by base insertion or deletion in small mononucleotide repeats in the coding region contributes to cancer development and progression.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Alterations of DNA mismatch repair proteins and microsatellite instability levels in gastric cancer cell lines

Yao

Hong

Kim

et al. 2004

Laboratory Investigation

View full text Add to dashboard Cite

Alterations in DNA mismatch repair (MMR) proteins result in microsatellite instability (MSI), increased mutation accumulation at target genes and cancer development. About one-third of gastric cancers display high-level microsatellite instability (MSI-High) and low-level microsatellite instability (MSI-Low) is frequently detected. To determine whether variations in the levels of MMR proteins or mutations in the main DNA MMR genes are associated with MSI-Low and MSI-High in gastric cancer cell lines, the MSI status (MSI-High, MSI-Low or MSStable (MSS)) of 14 gastric cancer lines was determined using multiple clone analysis with a panel of five microsatellite markers. Protein levels of hMLH1, hMSH2, hMSH6, hPMS2 and hPMS1 were determined by Western blot. Sequence analysis of hMLH1 and hMSH2 was performed and the methylation status of the hMLH1 promoter was examined. The cell lines SNU1 and SNU638 showed MSI-High, decreased to essentially absent hMLH1 and hPMS2 and reduced hPMS1 and hMSH6 protein levels. The hMLH1 promoter region was hypermethylated in SNU638 cells. The MKN28, MKN87, KATOIII and SNU601 cell lines showed MSI-Low. The MMR protein levels of cells with MSI-Low status was similar to the levels detected in MSS cells. A marked decrease in the expression levels of MutL MMR proteins (hMLH1, hPMS2 and hPMS1) is associated with high levels of MSI mutations in gastric cancer cells. Gastric cancer cell lines with MSI-Low status do not show significant changes in the levels of the main DNA MMR proteins or mutations in the DNA mismatch repair genes hMSH2 and hMLH1. These well-characterized gastric cancer cell lines are a valuable resource to further our understanding of DNA MMR deficiency in cancer development, progression and prognosis. Keywords: gastric cancer cell lines; microsatellite instability; mismatch repair genes; stomach cancer; mutations The DNA mismatch repair (MMR) system is essential for replication fidelity, correcting DNA base mismatches left uncorrected by DNA polymerase. Efficient DNA mismatch repair requires the combined function of MutL (hMLH1-hPMS2 and hMLH1-hMLH3) and MutS (hMSH2-hMSH6 and hMSH2-hMSH3) heterodimers.

show abstract

Medicinal Chemistry: Defining Itself

Metternich

Tarzia

2008

ChemMedChem

View full text Add to dashboard Cite

Identification of a Second MutL DNA Mismatch Repair Complex (hPMS1 and hMLH1) in Human Epithelial Cells

Cited by 59 publications

References 25 publications

Increased expression of survivin in gastric cancer patients and in first degree relatives

Increased expression of survivin in gastric cancer patients and in first degree relatives

Alterations of DNA mismatch repair proteins and microsatellite instability levels in gastric cancer cell lines

Medicinal Chemistry: Defining Itself

Contact Info

Product

Resources

About