| 吴丽娜,朴春南,田梅,阮健磊,刘建香.抑制γ-突触核蛋白表达对乳腺癌T47D细胞放射敏感性的影响[J].中华放射医学与防护杂志,2017,37(1):19-23 |
| 抑制γ-突触核蛋白表达对乳腺癌T47D细胞放射敏感性的影响 |
| Effect of interference of SNCG gene on radiosensitivity of breast cancer T47D cells |
| 投稿时间：2016-08-24 |
| DOI：10.3760/cma.j.issn.0254-5098.2017.01.004 |
| 中文关键词: 乳腺癌 γ-突触核蛋白 放射敏感性 基因治疗 |
| 英文关键词:Breast cancer γ-synuclein(SNCG) Radiosensitivity Gene therapy |
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| 中文摘要: |
| 目的探讨γ突触核蛋白（γ-synuclein，SNCG）对乳腺癌细胞辐射敏感性的影响。方法合成SNCG的siRNA干扰序列并转染T47D细胞。用RT-PCR和Western blot法分别从基因水平和蛋白水平检测SNCG基因的表达。实验设SNCG siRNA干扰组（干扰组）、阴性对照组、空白对照组。各组细胞分别接受不同剂量γ射线照射，集落形成实验检测细胞放射敏感性，CCK-8实验检测细胞增殖能力变化，Western blot法检测AKT及mTOR磷酸化变化。结果 SNCG siRNA干扰组较空白对照组的SNCG基因和蛋白表达都明显下降。在4、6、8 Gy照射下干扰组的乳腺癌细胞数量明显低于未转染SNCG的对照组（t=5.449、8.882、21.503，P<0.05），其中6 Gy照射时，干扰组的细胞增殖能力在照后24、48、72 h均较其他对照组低（t=5.603、4.839、6.115，P<0.05），且干扰组AKT及mTOR磷酸化水平降低，而总的AKT及mTOR未见明显变化。结论抑制SNCG的表达可增强乳腺癌细胞对射线的放射敏感性，其机制可能与AKT信号通路被抑制有关。 |
| 英文摘要: |
| Objective To explore the role of γ-synuclein(SNCG) siRNA in the radiosensitivity of breast cancer T47D cells.Methods SNCG siRNA was synthesized according to the coding sequence of SNCG mRNA and then transiently transferred into T47D cell with lipofectamine. The expression of SNCG gene and protein was detected by RT-PCR and Western-blot, respectively. Cells were divided into three groups, SNCG siRNA interference group, negative control group and blank control group, which were irradiated with different doses of 60Co γ-rays. Cell radiosensitivity was evaluated by colony formation assay, cell proliferation was assayed by CCK-8 kit, and the protein expressions of phosphorylated-AKT and mTOR were detected by Western blot assay. Results Compared with blank control cells, the expressions of SNCG gene and protein in the SNCG siRNA transferred T47D cells were efficiently diminished. Cell colony formation results showed that, under 4, 6, 8 Gy irradiation, the cell survival of siRNA transfection group was lower than that of control group (t=5.449, 8.882, 21.503, P<0.05). CCK-8 experiments showed that the cell proliferation abilities of siRNA group at 24, 48, 72 h after 6 Gy irradiation were lower than those of control group (t=5.603, 4.839, 6.115, P<0.05). In addition, after 6 Gy irraddaition, the AKT and mTOR phosphorylation levels in the siRNA group were more obviously reduced compared with blank groups, but the total AKT and mTOR had no changes.Conclusions Transfection of SNCG siRNA can enhance the radiosensitivity of breast cancer cells probably by inhibiting p-AKT signal pathway. |
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