General

• The U-CH1 cell line had an immunocytochemical profile usually found in chordomas: co-expression of S-100 protein, vimentin, EMA, and cytokeratin. [1]
• U-CH1 and its parent tumor had almost the same CGH profile. [1]
• The mean apoptosis index of chordoma cells was significantly lower compared with that of notochordal cells (2.1% ± 1.6% vs. 3.8% ± 1.6%, P=0.03). The mean proliferation potential index of chordoma cells was significantly higher than in notochordal cells (6.8% ± 1.2% vs. 2.1% ± 1.2%, P = 0.01). [2]
• Microarray analysis, TaqMan RT-PCR, and Northern blot found up-regulation of miRNA-320 (hsa-miR-320c), miRNA-130 (hsa-miR-130b), miRNA-193 (hsa-miR-193a-5p), and miRNA-34 (hsa-miR-34a) in the U-CH1 and CH8 cell lines and in primary chordoma tissue. [3]
• Microarray analysis, TaqMan RT-PCR, and Northern blot found down-regulation of miRNA-27 (hsa-miR-27a), miRNA-23 (hsa-miR-23a), miRNA-572 (hsa-miR-572), miRNA-128 (hsa-miR-128), miRNA-1268 (hsa-miR-1268), miRNA-628 (hsa-miR-628-3p), miRNA-101 (hsa-miR-101), miRNA-26 (hsa-miR-26a), miRNA-1228 (hsa-miR-1228), miRNA-940 (hsa-miR-940), miRNA-133 (hsa-miR-133a and hsa-miR-133b), miRNA-139 (hsa-miR-139-5p), miRNA-1 (hsa-miR-1), miRNA-206 (hsa-miR-206), miRNA-933 (hsa-miR-933), miRNA-95 (hsa-miR-95) in the U-CH1 and CH8 cell lines and in primary chordoma tissue. [3]
• Microarray analysis found a drastic reduction in the expression of miRNA-1 (hsa-miR-1), miRNA-206 (hsa-miR-206), and miRNA-133 (hsa-miR-133a and hsa-miR-133b) in the U-CH1 and CH8 cell lines and in primary chordoma tissue. [3]
• Western blot analysis found overexpression of the miRNA-1 targets Met and HDAC4 in the chordoma cell lines U-CH1, CH8, and GB60, and in eight primary chordoma samples. [3]
• Transfecting U-CH1 cells with miRNA-1 led to dose- and time-dependent decrease in Met expression. This also led to dose-dependent inhibition of cell growth. [3]
• MTT assay found that cells from the U-CH1 chordoma cell line proliferated more rapidly in DMEM media and less rapidly in RPMI 1640. [4]
• MTT and Hoechst assays indicated that growth of cells from the CH8, GB60, and U-CH1 chordoma cell lines was unaffected by oxygen and glucose concentrations. [4]
• Cell numeration and Hoechst assays confirmed improved growth of cells from the CH8, GB60, and U-CH1 chordoma cell lines when cultured on collagen substrate. [4]
• Microscopic observation of cultures from the novel 3D model of the CH8, GB60, and U-CH1 chordoma cell lines found that the chordoma cells became spheroid upon being seeded in the 3D culture. These cells formed clusters after 5 – 6 days and then developed into acini-like spheroids. Western blot analysis and immunofluorescence staining confirmed expression of the chordoma cell markers cytokeratin and vimentin. CH8 cells tended to proliferate more rapidly than GB60 or U-CH1 cells. [4]
• MTT assay found that cells from the CH8, GB60, and U-CH1 chordoma cell lines were more sensitive to the chemotherapeutic drugs doxorubicin, yondelis, zalypsis, and cisplatin than to methotrexate or paclitaxel. [4]
• Treatment of U-CH1 cells with PI-103 in conjunction with either doxorubicin or cisplatin led to synergistic increase in apoptosis. [5]
• FISH analysis confirmed high polysomy of chromosome 6 in the U-CH1 chordoma cell line. [6]
• Tyrphostin inhibited proliferation of U-CH1 cells in a dose-dependent manner, as assessed by an MTS assay. As cells slowed in proliferation, they also underwent morphological changes. [7]

Brachyury (T)

• Lentiviral-delivered shRNA caused knockdown of brachyury in U-CH1 cell lines and resulted in cells with abnormal shapes, spindling, flattening, and extensive branching, which first appeared four days after infection and was progressive in nature. When the same experiment was performed on 293T and HeLa cell lines, no morphological or proliferative changes were observed. [8]
• 65% of U-CH1 cells with knocked down brachyury stained positive for senescence-associated beta galactosidase, and this growth arrest was confirmed through observation of a very low cell density in brachyury knockdown cultures at two weeks after shRNA infection. [8]
• Lentivirus-introduced hairpin knockdown of T in U-CH1 cells caused premature senescence and growth arrest, as indicated by spindling, flattening, extensive branching, and positive staining for beta galactosidase in 65% of cells. [6]

Receptor Tyrosine Kinase

• Application of the EGFR tyrosine kinase inhibitor tyrphostin to U-CH1 cells resulted in spindling, reduced cell size, and decreased cell density. [8]
• MTS indirect proliferation assay showed that the EGFR tyrosine kinase inhibitor tyrphostin inhibited growth of U-CH1 cells in a dose-dependent manner, resulting in a progressive reduction of cell number over 7 days. Removal of tyrphostin showed a recovery of cell number after 4 days. [8]
• EGFR phosphorylation was reduced after treating U-CH1 cells with tyrphostin in p-EGFR-specific membranes. [8]
• Immunohistochemistry showed that treatment of U-CH1 cells with tyrphostin reduced phosphorylation of EGFR but had no effect on brachyury expression. [7]

Cytokine Receptor

• Western blot analysis showed expression of MCL-1, Bcl-xL, pStat3, Stat3, pSrc, and Src, components of the Src/Stat3 pathway, in 7 human chordoma samples and in the chordoma cell lines CH-8, U-CH1, and GB-60. [9]
• Western blot analysis showed that treatment of the chordoma cell lines CH-8, U-CH1, and GB-60 with 2-cyano-3,12-dioxooleana-1,9 (11)-dien-28-oic acid-methyl ester (CDDO-Me) inhibited expression of Stat3, pStat3, Src, pSrc, Bcl-xL, and MCL-1, and led to PARP cleavage. Treatment also resulted in reduced growth among the cell lines. [9]
• Cytotoxicity assay revealed synergistic inhibition of growth in cultures from the chordoma cells lines CH-8, U-CH1, and GB-60 upon treatment with CDDO-Me and either doxorubicin or cisplatin. [9]
• Cell numeration analysis demonstrated a decrease in the growth of CH-8, U-CH1, and GB-60 chordoma cells grown in 3D culture after treatment with CDDO-Me. [9]
• Immunohistochemistry showed inhibition of expression of pSrc and pStat3 in the CH-8, U-CH1, and GB-60 cell lines after treatment with CDDO-Me in 3D culture. [9]
• Subcutaneous injection of U-CH1 cells into NOD/SCID/interluekin -null mice resulted in xenografts after 10 weeks. [6]
• Immunohistochemistry found the U-CH1 cell line to be positive for EGFR and p-EGFR. [7]

PI3K

• DNA investigation yielded a nonsense mutation (G2045T) in exon 15 of the TSC1 gene of a male, leading to translation termination in codon 609. The same mutation was identified in the remaining fixed tumor of his son’s chordoma. [10]
• The U-CH1 cell line showed immunoreactivity for p-AKT, p-TSC2, p-mTOR, p-RPS6, p-S6K, p-4E-BP1, and eIF-4E. [8]
• Expression of p-AKT and p-ERK1/2 was shown through Western blot analysis to decrease significantly in U-CH1 cells after treatment with tyrphostin. [8]
• Treatment of U-CH1 cells with the chemotherapy agent PI-103 resulted in dose-dependent inhibition of phosphorylation of AKT and mTOR, and inhibition of the anti-apoptosis protein BcL-XL, as shown by Western blot analysis. This led to both decreased proliferation and increased apoptosis in U-CH1 cells. [5]

References