Kudawara Ikuo, Araki Nobuhito, Myoui Akira, Kato Yoichi, Uchida Atsumasa, Yoshikawa Hideki
Department of Orthopaedic Surgery, Osaka National Hospital, Osaka 2-1-14 Hoenzaka, Chuo-ku, 540-0006 Osaka, Japan.
Virchows Arch. 2004 Jun;444(6):577-86. doi: 10.1007/s00428-004-1020-6. Epub 2004 Apr 29.
In the present study, we investigated chondrocytic characterization for newly established human chondrosarcoma cell lines. A chondrosarcoma cell line, HCS-TG, was established by the implantation of grade-2 human chondrosarcoma into athymic mice. Cloning of HCS-TG cells from passage 17 was performed. After cell cloning, two clonal-cell lines (HCS-TG C3 and E2) with good proliferative activities were obtained. These cell lines in monolayer culture retained a polygonal morphology. Their doubling times were 68 h and 45 h, respectively. mRNA expressions of type-I, -II, -X, and -XI collagens and aggrecan core protein were detected on reverse-transcription polymerase chain reaction. Protein expression of type-II collagen was confirmed in each cell line using Western blotting. However, there was no expression of type-I collagen. Moreover, gelatin zymography revealed that both cell lines produced extracellular matrices with matrix metalloproteinases 2 and 9. The parental HCS-TG cells had tumorigenicity in athymic mice; however, C3 and E2 were not tumorigenic. New clonal-cell lines HCS-TG C3 and E2 derived from human chondrosarcoma are morphologically chondrocytic in serial monolayer cultures and express chondrocytic phenotypes.
在本研究中,我们对新建立的人软骨肉瘤细胞系进行了软骨细胞特性研究。通过将2级人软骨肉瘤植入无胸腺小鼠体内,建立了软骨肉瘤细胞系HCS-TG。对第17代的HCS-TG细胞进行克隆。细胞克隆后,获得了两个具有良好增殖活性的克隆细胞系(HCS-TG C3和E2)。这些单层培养的细胞系保持多边形形态。它们的倍增时间分别为68小时和45小时。通过逆转录聚合酶链反应检测了I型、II型、X型和XI型胶原蛋白以及聚集蛋白聚糖核心蛋白的mRNA表达。使用蛋白质印迹法在每个细胞系中证实了II型胶原蛋白的蛋白表达。然而,未检测到I型胶原蛋白的表达。此外,明胶酶谱分析表明,这两个细胞系都产生含有基质金属蛋白酶2和9的细胞外基质。亲代HCS-TG细胞在无胸腺小鼠中具有致瘤性;然而,C3和E2没有致瘤性。源自人软骨肉瘤的新克隆细胞系HCS-TG C3和E2在连续单层培养中形态上呈软骨细胞样,并表达软骨细胞表型。