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In colon cancer cell lines, Wnt signaling can increase transcription of VEGFA, and elevated levels of VEGFA have been detected in intestinal polyps of APCmin/+ mice and in human colon cancers relative to matched normal colon tissues [19].
At the time, a section of matched normal colon tissue was extracted adjacent to the tumour site.
We then compared matched normal colon tissue, primary tumour and colorectal liver metastasis from five individual patients.
The presence of altered/additional bands that resulted from the tumour PCR-amplified product, when compared with the matched normal colon PCR product, indicated MSI.
Two examples of CEACAM6 staining for matched normal colon tissue, primary colon tumor, and liver metastases are shown in Figure 8.
39 colorectal cancer samples were also examined with 35 exhibiting a greater than two-fold increase in G-CSFR mRNA levels over the matched normal colon tissue.
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We sequenced tumor and matching normal colon tissues from two patients with high grade adenocarcinoma of the colon (G3), patient 1 with a microsatellite instable and patient 2 with a microsatellite stable tumor (Table 1, Figure S1).
COBRA showed that the CGIs encompassing the promoter and/or coding region of DCC, TBX5, TBX18, SLIT1 were methylated in primary colorectal tumors but not in matching normal colon tissues whereas GNA11 was methylated in both.
Next, we extended our study to analyze the methylation status of a selected group of genes (DCC, TBX18, TBX5, SLIT1 and GNA11) in several primary human colorectal tumors and matching normal colon tissues by COBRA.
The CGI spanning promoter/exon 1 of DCC was methylated at Taq I site in 8 out of 10 tumors as demonstrated by almost complete digestion with Taq I whereas it was methylated in matching normal colon tissues only in sample #8 and #10 (Figure 3.i).
To elucidate the reason of gene expression reduction, we isolated targeted human colon cancer cells and matched normal colonic epithelial cells from colorectal cancer patients with low tumor-SBP1 protein and mRNA expression [10] for SBP1 promoter methylation analysis.
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