Park K H, Rha S Y, Kim C H, Kim T S, Yoo N C, Kim J H, Roh J K, Noh S H, Min J S, Lee K S, Kim B S, Chung H C
Yonsei Cancer Research Institute, Yonsei University College of Medicine, Seoul, Korea.
Int J Oncol. 1998 Sep;13(3):489-95. doi: 10.3892/ijo.13.3.489.
For the cancer cells which have overcome the second mitotic clock (M2), activated telomerase is essential and used as another marker of immortality. Many trials had been initiated to target telomerase, which is known to be specific to tumors. To determine the best in vitro cell system for testing the efficacy of telomerase inhibitors, we evaluated the telomerase activity of various cancer cell lines and measured their telomere lengths. We also treated some cancer cell lines with adriamycin and measured the changes of telomerase activity. Telomerase activity was evaluated in various cell lines with the TRAP (telomeric repeat amplification protocol) assay. Telomerase activity was calculated and translated into arbitrary units by computer-assisted densitometry with the control of telomerase activity in the 293 control cell line. Also, terminal restriction fragment lengths were measured using Southern blotting. We also measured telomerase activity and telomere lengths in 11 benign breast tumor tissues and 19 paired stomach cancer and normal tissues. Cancer cell lines treated with adriamycin we evaluated for changes of telomerase activity and the cell proliferation by MTT assay and dye exclusion test. Telomerase activity of cell lines was 95.3 24.1 unit with a range of 27.6-129.6 unit, while the telomere lengths of those cell lines were variable from 5.0 to 10.4 kbp with a median of 6 kbp. In 11 cancer cell lines which were not yet firmly established, we could not detect any telomerase activity. Low telomerase activity was detected in only 2 benign tumor tissues of breast with a median telomere length of 8.8 (7-10.5) kbp. Among paired 19 gastric cancer and normal tissues, only 7 cancer tissues showed weak telomerase activity. After adriamycin treatment, telomerase activity in YCC-S-1, YCC-S-3, MCF-7 and MCF-7/ADR was decreased in accordance with the changes of the cell numbers. Telomerase is specific to cancer tissues and is expressed differently from organ to organ. Telomerase activity by TRAP assay could be used as a chemosensitivity assay.
对于已克服第二个有丝分裂时钟(M2)的癌细胞,活化的端粒酶至关重要,并被用作永生的另一个标志物。已经开展了许多针对端粒酶的试验,端粒酶已知对肿瘤具有特异性。为了确定用于测试端粒酶抑制剂疗效的最佳体外细胞系统,我们评估了各种癌细胞系的端粒酶活性并测量了它们的端粒长度。我们还用阿霉素处理了一些癌细胞系并测量了端粒酶活性的变化。用端粒重复序列扩增法(TRAP)测定评估各种细胞系中的端粒酶活性。通过计算机辅助光密度测定法计算端粒酶活性并将其转化为任意单位,并以293对照细胞系中的端粒酶活性作为对照。此外,使用Southern印迹法测量末端限制片段长度。我们还测量了11个良性乳腺肿瘤组织以及19对胃癌组织和正常组织中的端粒酶活性和端粒长度。我们通过MTT法和染料排除试验评估了用阿霉素处理的癌细胞系中端粒酶活性的变化和细胞增殖情况。细胞系的端粒酶活性为95.3±24.1单位,范围为27.6 - 129.6单位,而这些细胞系的端粒长度在5.0至10.4 kbp之间变化,中位数为6 kbp。在11个尚未稳固建立的癌细胞系中,我们未检测到任何端粒酶活性。仅在2个良性乳腺肿瘤组织中检测到低端粒酶活性,端粒长度中位数为8.8(7 - 10.5)kbp。在19对配对的胃癌组织和正常组织中,只有7个癌组织显示出弱端粒酶活性。阿霉素处理后,YCC - S - 1、YCC - S - 3、MCF - 7和MCF - 7/ADR中的端粒酶活性随细胞数量的变化而降低。端粒酶对癌组织具有特异性,并且在不同器官中的表达有所不同。通过TRAP测定的端粒酶活性可作为一种化学敏感性测定方法。