Establishment and characterization of a novel osteosarcoma cell line: CHOS

Abstract

Osteosarcoma has a well-recognized bimodal distribution, with the first peak in adolescence and another in the elderly age-group. The elderly patients have different clinical features and a poorer prognosis as compared to adolescents. To better understand the biological features of osteosarcoma in the elderly population, we established a new human osteosarcoma cell line from a 58-year-old man with primary chondroblastic osteosarcoma. After 6 months of continuous culture in vitro for over 50 passages, an immortalized cell line CHOS was established. The cell line was well-characterized by cytogenetic, biomarker, functional, and histological analyses. The CHOS cells exhibited a spindle-shaped morphology and a doubling time of 36 h. Cytogenetic analysis of CHOS cells revealed the loss of chromosome Y and the gain of chromosome 12. Quantitative real-time polymerase chain reaction (RT-PCR), Western blotting and/or immunofluorescence revealed the expression of chondroblastic, mesenchymal and tumor metastasis markers in the CHOS cells. Compared with the osteosarcoma cell line, the CHOS cells were found to be more sensitive to cisplatin and doxorubicin, but were resistant to methotrexate. The cell line was highly tumorigenic and maintained the histological characteristics and invasive nature of the original tumor. Furthermore, on immunohistochemical analysis, the xenografts and metastases were found to co-express collagen II, aggrecan, vimentin and S100A4 that resembled the original tumor cells. Our results indicate, the potential of CHOS cell line to serve as a useful tool for further studies on the molecular biology of osteosarcoma, especially in the elderly patients. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:2116-2125, 2016.

Establishment and Characterization of a Recurrent Osteosarcoma Cell Line: OSA 1777

“…While most of the previously established OSA cell lines including HOS, MNNG‐HOS, KHOS, U2OS, and MG63 were derived from the primary tumor, this OSA 1777 cell line was established from the recurrent conventional OSA (Supplementary Table S1) . As there are no specific proteins or genetic markers for OSA confirmation, the principal diagnostic tool that remains is histologic examination and patient history.…”

“…While most of the previously established OSA cell lines including HOS, MNNG-HOS, KHOS, U2OS, and MG63 were derived from the primary tumor, this OSA 1777 cell line was established from the recurrent conventional OSA ( Supplementary Table S1). [7][8][9][10][11][12][13][14][15][16] As there are no specific proteins or genetic markers for OSA confirmation, the principal diagnostic tool that remains is histologic examination and patient history. As shown in Figure 1, the H&E stain of recurrent tumor tissues was consistent with highgrade OSA and showed typical anaplastic osteoblasts elaborating osteoid matrix; there was 30% tumor necrosis after second-line chemotherapeutic treatment.…”

Mitochondrial Pathway Is Involved in Advanced Glycation End Products-Induced Apoptosis of Rabbit Annulus Fibrosus Cells

“…Researchers showed that cardiomyocyte viability decreased in a dose-dependent manner in diabetic cardiomyopathy after treatment with AGEs. 32 In addition, Liu et al 28 reported that AGEs significantly decreased chondrocyte viability. Similarly, studies by other researchers have shown that decreases in chondrocyte viability occurred after treatment with AGEs.…”

“…Cell viability was detected by Cell Counting Kit-8 (CCK-8, Dojindo, Japan), as described previously. 28 The cell densities were 5 × 10 3 cells/well for CCK-8. After being seeded in 96-well culture plates, AF cells were treated with AGEs at various concentrations for 3d.…”

The dual role of poly(ADP-ribose) polymerase-1 in modulating parthanatos and autophagy under oxidative stress in rat cochlear marginal cells of the stria vascularis

“…Equal amounts of protein samples were subjected to 10–12% SDS-PAGE separation and then transferred to the polyvinylidene difluoride membrane (Millipore, Boston, MA, USA). Subsequent procedures were performed as described previously [38] . The following antibodies were used: rabbit anti-PARP-1 (1: 500, Santa Cruz, CA, USA), rabbit anti-AIF (1:1000, Abcam), rabbit anti-LC-3 (1:1000, Sigma-Aldrich), rabbit anti-p62 (1:1000, Cell Signaling Technology, MA, USA), mouse anti-PAR (1:1000, Enzo Life Sciences), mouse anti-β-actin (1:5000, Abbkine), rabbit anti-COX Ⅳ (1:1000, Proteintech, Wuhan, China), mouse anti-Histone H3 (1:3000, Abbkine), HRP-conjugated secondary antibodies (1:5000, Abbkine).…”