人下颌骨骨肉瘤细胞系HMOS-99的生物学特性研究

The biological characteristics of human mandibular osteosarcoma cell line HMOS-99

目的:检测人下颌骨骨肉瘤细胞系HMOS-99的细胞生物学特征。方法:通过生长曲线计算倍增时间、测定集落形成率、分析细胞周期等探讨细胞生长增殖特性;分析染色体核型;免疫组化方法检测CKpan、Vim、BMP-2、COⅠ、OC、BSP、OPN的表达;细胞化学染色试剂盒检测ALP的表达;皮下接种免疫缺陷小鼠,检测HMOS-99细胞的成瘤性。结果:HMOS-99细胞倍增时间为42.98h,平板集落形成率为58.5%;细胞周期测定S期细胞占28.44%,并表现为异倍体细胞;核型分析显示细胞呈亚三倍体核型;CKpan(-)、Vim(+)、BMP-2(+)、COⅠ(+)、OC(+)、BSP(+)、OPN(+)、ALP(+);免疫缺陷小鼠皮下接种未见肿瘤形成。结论:人下颌骨骨肉瘤细胞系HMOS-99的生物学特性符合骨肉瘤细胞,接种免疫缺陷小鼠无肿瘤生成。

PURPOSE: To study the biological characteristics of human mandibular osteosarcoma cell line HMOS-99. METHODS: Growth curve of the cells was described by cell counting. The clony formation was detected by clone accounting. The cell cycle phase of the cultured cells was analyzed by flow cytometer. Karyotypic analysis was done to find out the change of the transmissibility of the cells. Immunohistochemical staining was used to detect the expression of CKpan, Vim, BMP-2, COⅠ, OC, OSP and OPN. The cytochemical staining kit was used to find out the existing of ALP in the cultured cells. The tumorigenicity of the cultured cells was examined by transplanting the cells subcutaneously to the nude mice and SCID mice. RESULTS: According to the growth curve, It was found that the cells pass through three stages: latent phase, log phase and plat phase. DT=42.98h. The clony formation was 58.5%. Cell cycle phase analysis showed that there were 59.55% cells in G1 stage, 12.95% cells in G2 stage and 28.44% cells in S stage. Most cells were euploid. Karyotipic analysis of one hundred and ten cells of the 13th, 38th, 72th passage indicated that the number of chromosome was between 49 and 60, as hypotriploid. The immunohistochemical and cytochemical staining showed that the cells expressed Vim, BMP-2, COⅠ, OC, OSP, OPN and ALP, but no CKpan. The results indicated the cells were expected from the mesodermic origin of osteoblasts. Six four-week-old nude mice, eleven SCID mice were used for transplantation. Subcutaneously injection was carried out in, 12 points and 11 points, respectively. The sites of injection were inspected once a week for four months. There was no tumor growth. CONCLUSIONS: The established cell line has the biological characteristics of osteosarcoma cells and will provide a good experiment model for the studies of osteosarcoma.