| Liu Zhengchun,Cai Rui,Zhang Kaili,et al.Effects of lentivirus-mediated DKC1 gene silence on radiosensitivity of human cervical cancer HeLa cells[J].Chinese Journal of Radiological Medicine and Protection,2020,40(8):590-594 |
| Effects of lentivirus-mediated DKC1 gene silence on radiosensitivity of human cervical cancer HeLa cells |
| Received:December 31, 2019 |
| DOI：10.3760/cma.j.issn.0254-5098.2020.08.003 |
| KeyWords:DKC1 gene Cervical cancer HeLa cells Radiosensitivity Lentivirus shRNA |
| FundProject:广西自然科学基金面上项目（2016GXNSFAA380306） |
| Author Name | Affiliation | E-mail | | Liu Zhengchun | Department of Radiation Oncology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China | | | Cai Rui | Department of Radiation Oncology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China | | | Zhang Kaili | Department of Radiation Oncology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China | | | Ruan Guozhu | Department of Radiation Oncology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China | | | Liu Meilian | Department of Radiation Oncology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China | liu.meilian@163.com |
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| Abstract:: |
| Objective To investigate the effect of decreasing DKC1 gene expression on radiosensitivity of HeLa cells. Methods A cell model with low expression of DKC1 gene was established by shRNA technology with lentivirus as vector, and the interference efficiency was verified by RT-PCR and Western blot assay. Cells were divided into two groups of interference (Lv-shDKC1) and its negative control. Telomerase activity was detected by TRAP-ELISA, and telomere length was measured by Real-time PCR. Cell survival was obtained through clone formation assay and fitted by multi-target single-hit model, and radiobiological parameters (D0, Dq, N, SF2) and radiosensitization ratio (SER) were calculated. Results After DKC1 interfering, the expression levels of mRNA and protein of DKC1 in HeLa cells were significantly decreased by (71.330±4.112)% (t=25.53,P<0.05) and (35.520±3.804)% (t=4.833, P<0.05), respectively. Compared with the blank control group and negative control group, the telomerase activity of Lv-shDKC1 group decreased significantly from 0.900±0.044 and 0.897±0.031 to 0.713±0.021 (F=31.44, P<0.05), the relative telomere length was significantly decreased from 4.233±0.306 and 4.633±0.379 to 2.667±0.404 (F=39.15, P<0.05). The telomerase activity and relative telomere length of blank control group and Lv-shDKC1 negative control group had no significant difference(P>0.05). SF2 in the interference group (0.571±0.006) was significantly lower than that of the blank control group (0.861±0.009) and the Lv-shDKC1 negative control group (0.807±0.002) (F=1812, P<0.05), and the radiosensitization ratio (SER) of shDKC1 interference was 1.508. Conclusions Downregulation of DKC1 in human cervical cancer HeLa cells enhances the radiosensitivity through inhibiting the activity of telomerase and shortening the length of telomere. DKC1 gene may become a new target of radiosensitization. |
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