| QIANG Wei-guang,WU Qin-qin,YANG Lei,et al.Influence of TPP1 siRNA on the radiosensitivity of telomerase-negative human osteosarcoma cells U2OS[J].Chinese Journal of Radiological Medicine and Protection,2013,33(6):579-583 |
| Influence of TPP1 siRNA on the radiosensitivity of telomerase-negative human osteosarcoma cells U2OS |
| Received:March 06, 2013 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.06.003 |
| KeyWords:RNA interference TPP1 Telomere Telomere length Radiosensitivity |
| FundProject:国家自然科学基金（81071825） |
| Author Name | Affiliation | E-mail | | QIANG Wei-guang | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | WU Qin-qin | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | YANG Lei | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | LIU Zheng-chun | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | JIANG Huan-gang | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | ZHOU Fu-xiang | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | XIE Cong-hua | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | | | ZHOU Yun-feng | Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Clinical Cancer Study Center of Hubei Province, Key Laboratory of Tumor Biological Behavior of Hubei Province, Wuhan 430071, China | yfzhouwhu@163.com |
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| Abstract:: |
| Objective To observe the changes of telomere length and radiosensitivity of U2OS cells after its TPP1 gene is suppressed by small interfering RNA (siRNA). Methods According to the coding sequence of TPP1 mRNA, the interference sequence of TPP1-siRNA was designed. The U2OS cells were transfected with TPP1-siRNA by lipofectamine. The expression of TPP1 was detected by RT-PCR and Western blot, the telomere length was measured by Real-time PCR. The cell apoptosis was analyzed by flow cytometry, and the cell radiosensitivity was determined by clone formation assay. Results The inhibition rates of TPP1 gene and protein expressions in the TPP1-siRNA interference group were (65.70±2.10)% and (74.80±3.20)%, respectively. The results of Real-time PCR showed that the telomere length of the TPP1-siRNA interference group (0.99±0.07) was significantly shorter than that of the negative group (1.64±0.05) and the blank control group (1.55±0.05) (F=113.68,P<0.01). Compared with the negative group, the telomere length of the control group didn't change significantly (P=0.09). The apoptosis rate of TPP1-siRNA interference group (13.67±0.85)% was higher than those of negative control group (8.59±0.38)% and blank control group (8.18±0.21)% (F=92.32,P<0.01). U2OS cells with silenced TPP1 (0.6074±0.0115) had a SF2 lower than that of control group (0.8062±0.0056) (F=246.45,P<0.01), and the SER was 1.33. Conclusions Silencing of TPP1 in U2OS cells could shorten the telomere length and enhance the radiosensitivity. The telomere shortening caused by TPP1-siRNA is likely to be one of the reasons of radiosensitization. |
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