Identification of cell surface marker candidates on SV-T2 cells using DNA microarray on DLC-coated glass

Cell type dependent endocytic internalization of ErbB2 with an artificial peptide ligand that binds to ErbB2

“…Real-time quantitative PCR (RT-qPCR) was carried out on the genes of erbB1, erbB2, erbB3, erbB4 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) following the procedure previously described by Tuoya et al (2005). Briefly, 2 ng of internal control RNA was mixed with 5 mg of total RNA from each cell line, and reverse transcribed for single-strand cDNAs using oligo(dT) 18 primer.…”

Identification of TM9SF2 as a candidate of the cell surface marker common to breast carcinoma cells

“…Simultaneously, the probes complementary to the control RNAs described above were designed. Each probe was synthesized as 60-mer oligonucleotide with NH 2 group at the 5' end and spotted in duplicate onto DLC-coated glass slides with DNA spotting tool, SPBI0 TM (Hitachi software engineering, Japan) as previously described by Tuoya et al [9,10] Microarray analysis Prior to cDNA synthesis against the total RNA from each cell line, each control RNA (0.25 ng for β-gal, 0.5 ng for Neo, 1 ng for CAT, 2 ng for GFP and 4 ng for β-lac) was supplemented to 20 μg of each total RNA. Fluorescent-labeled cDNAs were prepared by Superscript II reverse transcriptase (Invitrogen) with oligodT primer from the RNA mixtures in the presence of amino-allyl-dUTP followed by the coupling of Cy3 dye (Ambion, TX).…”

“…Data fi ltering and sSOM analysis As described previously [9,10] , the intensity of each signal of gene expression levels was calculated as relative fl uorescent intensity (RFI), which is the percentage of the fl uorescent intensity of each gene when the signal of the control RNA of GFP is considered to be 100%. In order to eliminate genes, of which expression did not change significantly between carcinoma derived cell lines and normal cell line, we evaluated the scores for each gene by a fi ltering formula |A−G| − V, where "A", "G", and "V" denote the expression level of a gene in normal cell line Hs 578Bst, the average expression level of the gene in the eight breast carcinoma cell lines, and the standard deviation of the gene expression level among the eight breast carcinoma cell lines, respectively.…”

“…This heterogeneity of cell surface protein expression makes these molecules useful markers for tumor specific targeting [5][6][7] since cell-surface proteins are considered to be more accessible to immune mechanisms and drug delivery systems [8] . Recently we have developed DNA microarrays carrying the probes for the genes coding cell surface proteins [8][9][10] . We consider this design is highly effi cient for downstream procedure of screening for cell surface markers.…”

Engineered bio-nanocapsules, the selective vector for drug delivery system

“…Due to the specific affinity to the antibody, the specific tropism of the engineered BNC could be easily altered, depending on the antibody displayed on the surface of the BNC. Recently, we successfully designed a DNA microarray for the screening of cell surface markers (40). In the screening of the cell surface marker of SV40 transformed mouse fibroblasts, we found that the expression of CD62L and IL-6 receptor genes were upregulated, in comparison to normal fibroblasts.…”