Schwartz H S, Juliao S F, Sciadini M F, Miller L K, Butler M G
Department of Orthopaedics and Rehabilitation, Vanderbilt University Medical Center, Nashville, Tennessee 37232-2550.
Cancer. 1995 Mar 1;75(5):1094-9. doi: 10.1002/1097-0142(19950301)75:5<1094::aid-cncr2820750507>3.0.co;2-b.
Benign giant cell tumor of bone (GCT) is a primary skeletal neoplasm with an unpredictable pattern of biologic aggressiveness and cytogenetic findings characterized by telomeric associations and telomeric reduction. The role of maintaining telomeric integrity is performed by telomerase. To determine if telomerase activity is present, cell extracts from fibroblasts and tumor cells from five patients with GCT were analyzed and compared with HeLa (a positive control cell line).
Telomerase activity was detected by visualizing the extension of radioactive telomeric repeats on DNA sequencing gels. Telomere reduction was assessed using southern blot analyses of the restriction enzyme Hinf I digested DNA with a radio-labeled telomere probe.
Telomerase or telomerase-like activity was detected in the cell extracts from HeLa and tumor cells. However, GCT telomerase activity varied and was less than that observed in HeLa, but no activity was detected from fibroblasts. In addition, telomere reduction was seen in DNA isolated from both HeLa and GCT but not in fibroblasts or age-matched controls.
Telomere reduction and telomerase activity may be oncogenic sustaining events required to maintain the transformed phenotype seen in GCT.
骨良性巨细胞瘤(GCT)是一种原发性骨肿瘤,其生物学侵袭模式不可预测,细胞遗传学表现为端粒关联和端粒缩短。端粒酶负责维持端粒完整性。为确定是否存在端粒酶活性,对5例GCT患者的成纤维细胞和肿瘤细胞的细胞提取物进行了分析,并与HeLa细胞(一种阳性对照细胞系)进行比较。
通过在DNA测序凝胶上观察放射性端粒重复序列的延伸来检测端粒酶活性。使用放射性标记的端粒探针,通过对经限制性内切酶Hinf I消化的DNA进行Southern印迹分析来评估端粒缩短情况。
在HeLa细胞和肿瘤细胞的细胞提取物中检测到端粒酶或端粒酶样活性。然而,GCT的端粒酶活性各不相同,且低于HeLa细胞中的活性,但在成纤维细胞中未检测到活性。此外,在从HeLa细胞和GCT中分离的DNA中观察到端粒缩短,但在成纤维细胞或年龄匹配的对照中未观察到。
端粒缩短和端粒酶活性可能是维持GCT中所见转化表型所需的致癌维持事件。