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首页> 外文期刊>World Journal of Gastroenterology >Effects of phosphorothioate anti-sense oligodeoxynucleotides on colorectal cancer cell growth and telomerase activity
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Effects of phosphorothioate anti-sense oligodeoxynucleotides on colorectal cancer cell growth and telomerase activity

机译：硫代磷酸酯反义寡脱氧核苷酸对结直肠癌细胞生长和端粒酶活性的影响

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AIM: To investigate the inhibitory effect of phosphorothioate anti-sense oligodeoxynucleotides (PASODN) on colorectal cancer LS-174T cells in vitro and the mechanism of inhibition of telomerase activity in these cells. METHODS: PASODN were used to infect LS-174T cells and block human telomerase RNA (hTR) through anti-sense technology. The inhibitory effect of PASODN was evaluated by colony-forming inhibition assay and growth curve. Changes of telomerase activity in LS-174T cells were detected by polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELJSA), and the level of apoptosis was analyzed by flow cytometry (FCM) assay. RESULTS: PASODN showed a dose and time-dependent inhibition of cell proliferation. The optimal dosage of PASODN was 10 μmol/L. The colony-forming efficiency was 10.3% in PASODN group after 10 d, whereas that in phosphorothioate mis-sense oligodeoxynucleotides (PMSODN) group with the same concentration and in PBS group (blank control) was 49.1% and 50.7%, respectively. PCR-ELISA results indicated that telomerase activity in the PASODN group was obviously inhibited in comparison with in the control groups (P < 0.01, t = 3.317 and 3.241, t_(0.01(20)) = 2.845). Meanwhile, before the number of cells was decreased, the morphological changes were observed in the cells of PASODN group. The cells in PASODN group showed the apoptotic peak at 72 h after infection, whereas the control group did not show. CONCLUSION: Specific sequence oligonucleotides can inhibit telomerase activity and lead to cell apoptosis, suggesting a novel treatment strategy for malignant tumors induced by telomerase.
机译：目的：探讨硫代磷酸反义寡聚脱氧核苷酸（PASODN）对结直肠癌LS-174T细胞的体外抑制作用以及端粒酶活性的抑制机制。方法：使用PASODN通过反义技术感染LS-174T细胞并阻断人端粒酶RNA（hTR）。通过菌落形成抑制试验和生长曲线评价PASODN的抑制作用。聚合酶链反应-酶联免疫吸附法（PCR-ELJSA）检测LS-174T细胞端粒酶活性的变化，流式细胞仪（FCM）检测细胞凋亡水平。结果：PASODN显示出剂量和时间依赖性抑制细胞增殖。 PASODN的最佳剂量为10μmol/ L。在10 d后，PASODN组的菌落形成效率为10.3％，而相同浓度的硫代磷酸错义寡脱氧核苷酸（PMSODN）组和PBS组（空白对照组）的菌落形成效率分别为49.1％和50.7％。 PCR-ELISA结果表明，与对照组相比，PASODN组的端粒酶活性明显受到抑制（P <0.01，t = 3.317和3.241，t_（0.01（20））= 2.845）。同时，在PASODN组细胞减少之前，观察到细胞形态变化。 PASODN组细胞在感染后72 h出现凋亡高峰，而对照组则没有。结论：特异序列寡核苷酸可抑制端粒酶活性并导致细胞凋亡，为端粒酶诱导的恶性肿瘤的治疗提供了新的思路。

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