|
Objective To investigate the relationship between induced differentiation and regulation of telomerase activity in cultured osteosarcoma cells. Methods Osteosarcoma cells were treated by different doses of retinoic acid (RA) and dexamethasone (DEX). Telomerase activity and human telomerase RNA (hTR) of treated cells were detected by telomere repeat amplification protocol (TRAP) and by Northern blotting analysis. Results After treatment with RA or DEX, the growth of HOS cells was inhibited. Alkaline... Objective To investigate the relationship between induced differentiation and regulation of telomerase activity in cultured osteosarcoma cells. Methods Osteosarcoma cells were treated by different doses of retinoic acid (RA) and dexamethasone (DEX). Telomerase activity and human telomerase RNA (hTR) of treated cells were detected by telomere repeat amplification protocol (TRAP) and by Northern blotting analysis. Results After treatment with RA or DEX, the growth of HOS cells was inhibited. Alkaline phosphatase activity (differentiation marker of osteogenetic cells) of the cells increased significantly. Telomerase activity decreased with increasing treatment doses and times, but the expression level of hTR did not change obviously. Conclusions Both RA and DEX could inhibit cell growth and induce differentiation of HOS cells. Decreasing of telomerase activity could be considered as a marker of differentiation of HOS cells, but it does not correlate with the expression level of hTR. 【目的】探讨人骨肉瘤细胞分化与端粒酶活性调节之间的关系。【方法】用维甲酸和地塞米松诱导人骨肉瘤细胞分化 ,用端粒重复序列扩增法 (TRAP)检测端粒酶活性及用Northern印迹法检测人端粒酶RNA(RNA ,hTR)表达水平的改变。【结果】经维甲酸和地塞米松处理后 ,骨肉瘤细胞生长受到抑制 ,碱性磷酸酶活性升高 ,端粒酶活性明显下降 ,且与剂量 作用时间成负相关 ,但hTR表达水平并无改变。【结论】维甲酸和地塞米松有抑制骨肉瘤细胞增殖和诱导分化的作用 ,端粒酶活性下调可作为细胞分化指标 ,但与hTR表达水平不平行。 Objective To study the influence of anordrin (ANO),α-ANO, tamoxifen (TAM),cisplatin (CDDP) and all trans retinoic acid(ATRA) on the growth of Swarm rat chondrosarcoma (SRCS). Methods Effects of drugs on the growth of SRCS implanted into subrenal capsules of mice were studied in vivo. Calcium deposits in SRCS were revealed by von Kossa staining. Trypan blue exclusion method was used for examining the growth condition of SRCS cells cultured in vitro . The alterations of alkaline phosphatase (ALPase)... Objective To study the influence of anordrin (ANO),α-ANO, tamoxifen (TAM),cisplatin (CDDP) and all trans retinoic acid(ATRA) on the growth of Swarm rat chondrosarcoma (SRCS). Methods Effects of drugs on the growth of SRCS implanted into subrenal capsules of mice were studied in vivo. Calcium deposits in SRCS were revealed by von Kossa staining. Trypan blue exclusion method was used for examining the growth condition of SRCS cells cultured in vitro . The alterations of alkaline phosphatase (ALPase) activity of SRCS cells were observed with p nitrophenyl phosphate as substrate. o Cresolphthalein complexone colorimetry was employed to study the calcium content of SRCS cells. Results The tested drugs could inhibit the growth of SRCS implanted into subrenal capsules of mice at different degrees, and α-ANO caused 79.4% inhibition at the dosage 20 mg/kg. The IC 50 of CDDP,ATRA,TAM, α-ANO and ANO for SRCS cells after 96 h treatment wree 0 45, 1 47, 115 2, 161 7 and 241 5 μmol/L respectively. Calcium deposits were observed in SRCS treated by α-ANO and ATRA in vivo . α-ANO and ATRA also induced a significant elevation of ALPase activity and calcium content in SRCS cells in vitro for 96 h treatment. These phenomena were not observed in SRCS treated by CDDP and TAM. Conclusion Inducing SRCS toward differentiation and ossification was involved in the growth inhibition of SRCS caused by α-ANO and ATRA, but not by CDDP and TAM. The differentiation induction may be a new approach for clinical therapy of chondrosarcoma.$ 目的 研究双炔失碳酯 (ANO)及其 α异构体 (α-ANO)、三苯氧胺 (TAM)、顺铂 (CDDP)、全反式维甲酸(ATRA )对 Swarm大鼠软骨肉瘤 (SRCS)体内外生长的影响和机制。方法 观察药物 SRCS在小鼠肾囊腺下体内生长的影响 ;von Kossa法观察药物治疗对 SRCS中钙盐沉积的作用 ;用台盼蓝排染法观察了药物对 SRCS细胞体外生长的影响 ;以对硝基苯酚磷酸酯为底物研究药物对 SRCS细胞中碱性磷酸酯酶 (AL Pase)活性的影响 ;用邻甲酚酞络合酮比色法测定 SRCS细胞中的钙含量。结果 几种药物对 SRCS在小鼠肾囊膜下体内生长均有程度不一的抑制作用 ,α- ANO在 2 0 m g/ kg剂量下的生长抑制率为 79.4% ;CDDP、ATRA、TAM、α- ANO和 ANO作用 96 h对 SRCS细胞体外生长的 IC5 0 分别为 0 .45、1.47、115 .2、16 1.7和 2 41.5μmol/L ;α- ANO和 ATRA在体内可引起 SRCS中出现钙盐沉积 ;在体外作用 96 h可使 SRCS细胞中 AL Pase活性和钙含量显著性升高 ;而 CDDP和 ... 目的 研究双炔失碳酯 (ANO)及其 α异构体 (α-ANO)、三苯氧胺 (TAM)、顺铂 (CDDP)、全反式维甲酸(ATRA )对 Swarm大鼠软骨肉瘤 (SRCS)体内外生长的影响和机制。方法 观察药物 SRCS在小鼠肾囊腺下体内生长的影响 ;von Kossa法观察药物治疗对 SRCS中钙盐沉积的作用 ;用台盼蓝排染法观察了药物对 SRCS细胞体外生长的影响 ;以对硝基苯酚磷酸酯为底物研究药物对 SRCS细胞中碱性磷酸酯酶 (AL Pase)活性的影响 ;用邻甲酚酞络合酮比色法测定 SRCS细胞中的钙含量。结果 几种药物对 SRCS在小鼠肾囊膜下体内生长均有程度不一的抑制作用 ,α- ANO在 2 0 m g/ kg剂量下的生长抑制率为 79.4% ;CDDP、ATRA、TAM、α- ANO和 ANO作用 96 h对 SRCS细胞体外生长的 IC5 0 分别为 0 .45、1.47、115 .2、16 1.7和 2 41.5μmol/L ;α- ANO和 ATRA在体内可引起 SRCS中出现钙盐沉积 ;在体外作用 96 h可使 SRCS细胞中 AL Pase活性和钙含量显著性升高 ;而 CDDP和 TAM无上述现象。结论 α- ANO和 ATRA抑制 SRCS生长与其诱导 SRCS向成骨化方向分化有关 ,提示诱导分化治疗可能为临床治疗软骨肉瘤的新的有效途径。 Objective A rare huge desmoplastic fibroma on thoracic wall in 1 female case of 25 years old was resected, and the accompanying huge thoracic wall defect, ribs and soft tissues were repaired by tissue engineered bone and pedicled flap. The paper aims to explore the clinical results of early stage after operation. Methods Autogeneic bone marrow stromal cells (MSC) were obtained from bone marrow puncture of iliac bone and isolated and cultured according to the Houghton's methods, MSC were directively induced... Objective A rare huge desmoplastic fibroma on thoracic wall in 1 female case of 25 years old was resected, and the accompanying huge thoracic wall defect, ribs and soft tissues were repaired by tissue engineered bone and pedicled flap. The paper aims to explore the clinical results of early stage after operation. Methods Autogeneic bone marrow stromal cells (MSC) were obtained from bone marrow puncture of iliac bone and isolated and cultured according to the Houghton's methods, MSC were directively induced and differentiated to osteoblasts. Allogeneic ribs were made to the bio-derived bone scaffold materials after treatment of decell, deantigen, decalcification and dry freezing. 5×10 6/ml MSC were cocultured with the bio-derived bone for 6 days in vitro. After intact resection of tumor, the diaphragm flap was applied to repair pleural cavity, the three defect ribs were repaired by tissue engineered bone and the soft tissue defect was repaired by transfer of pedicled ipsilateral abdominal flaps. Results The patient recovered well with first intention. Followed up for 3 months, tissue engineered ribs were matured in vitro and the heart and pulmonary functions were improved markedly. Conclusion The tissue engineered bone constructed by autogeneic MSC is advantageous in individual treatment. 目的 应用组织工程技术构建的肋骨和带蒂皮瓣移位修复 1例 2 5岁女性患者胸壁巨大韧带样纤维瘤切除后 ,合并软组织、肋骨缺损的早期效果。方法 经髂骨穿刺抽取骨髓组织 ,按 Houghton法分离培养骨髓基质干细胞 ,经定向诱导分化为成骨样细胞 ;用同种异体肋骨经去细胞、去抗原、部分脱钙和冻干处理 ,制成保存天然框架的生物衍生骨支架 ,将 5× 10 6 /ml骨髓基质干细胞与骨支架材料联合培养 6天 ;待完整切除肿瘤后 ,用膈肌瓣修复胸膜腔 ,组织工程肋骨修复 3根肋骨 ,同侧带蒂侧腹壁皮瓣移位修复胸壁软组织缺损。结果 手术经过顺利 ,伤口 期愈合。术后 3个月随访 ,植入组织工程肋骨在体内逐渐发育成成熟肋骨 ,心肺功能显著改善。结论 应用自体骨髓基质干细胞构建的组织工程肋骨在个体化治疗中显示了优越性。
|