贵州医科大学学报

2020, v.45;No.236(05) 531-538

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阿立塞替通过靶向Aurora激酶A提高人非小细胞肺癌细胞的放射敏感性
Study on Alisertib Enhancing Radiosensitivity of Human Non-small Cell Lung Cancer Cells by Targeting Aurora Kinase A

李舜;杨智松;韩国征;李红艳;齐浩明;
LI Shun;YANG Zhisong;HAN Guozheng;LI Hongyan;QI Haoming;Daqing Longnan Hospital;Department of Traditional Chinese Medicine,the First Affiliated Hospital of Shihezi University;Department of Cardiovascular Medicine,Weinan Central Hospital;

摘要(Abstract):

目的:探讨阿立塞替(MLN8237)对人非小细胞肺癌放射敏感性的影响及作用机制。方法:用siPORT Neo FX转染试剂将阴性siRNA、si Aurora Kinase A分别转染NCI-H1975细胞阴性对照组(Si control)及靶向Aurora激酶A组(si Aurora Kinase A),未转染si PORT Neo FX的NCI-H1975细胞作为单纯对照组(Control);CCK8实验检测抑制Aurora激酶A的抑制剂(MLN82370)对NCI-H1975细胞生长的影响,细胞凋亡实验检测抑制Aurora激酶A及加入MLN8237对NCI-H1975细胞凋亡的影响,克隆形成实验检测了MLN8237对NCI-H1975细胞放射敏感性的影响影响,qRT-PCR实验评估了NCI-H1975细胞中p16及p21的表达,荧光素酶分析实验测定细胞周期相关转录因子(E2F)和应激活化蛋白激酶(JNK)的活性。结果:NCI-H1975细胞的转染效率达89. 91%,抑制Aurora激酶A及加入MLN8237可明显抑制NCI-H1975细胞的生长,差异有统计学意义(P <0. 01);细胞凋亡实验结果显示,抑制Aurora激酶A及加入MLN8237可明显提高NCI-H1975细胞的凋亡率;克隆形成实验显示,MLN8237能够显著降低照射后NCI-H1975细胞的克隆形成能力对NCI-H1975细胞有放射增敏效果,且随剂量增大其增敏效果越显著; qRT-PCR结果显示,加入MLN8237(Aurora激酶A抑制剂)照射后,p16及p21的表达明显上调;荧光素酶分析实验结果显示,抑制Aurora激酶A显著降低了E2F的活性,而JNK的活性显著提高,差异均有统计学意义(P <0. 01)。结论:阿立塞替能通过靶向Aurora激酶A,而抑制NCI-H1975细胞增殖及诱导其凋亡,从而提高NCI-H1975细胞的放射敏感性。
Objective: To investigate the effect of alisertib on the radiosensitivity of human non-small cell lung cancer by targeting Aurora kinase A. Methods: NCI-H1975 cells were transfected with si PORT NeoFX transfection reagent and divided into Si control group,Si Aurora Kinase A group; NciH1975 cells that were not transfected with si PORT NeoFX were used as the control group. The effects of inhibitors that inhibit Aurora kinase A( MLN82370) on NCI-H1975 cells growth were detected by CCK8 assay; the effects of Aurora kinase A inhibition and the addition of MLN8237 on the apoptosis of NCI-H1975 cells were detected by apoptosis assay; the effects of MLN8237 on radiosensitivity of NCIH1975 cells were examined by cloning formation experiment; the expression of p16 and p21 in NCIH1975 cells was evaluated by qRT-PCR; the activities of E2 F and JNK were determined by luciferase assay. Results: The transfection efficiency of NCI-H1975 cells was 89. 91%; CCK8 results showed the growth of NCI-H1975 cells were significantly inhibited by Aurora kinase A or adding the MLN8237,the difference was statistically significant( P < 0. 01). The results of apoptosis assay showed the apoptosis of NCI-H1975 cells were significantly increased by inhibiting of Aurora kinase A or adding MLN8237; The cloning formation experiments showed the cloning forming ability of NCIH1975 cells were significantly reduced with MLN8237 after irradiation,the radiosensitivity of NCIH1975 cells were significantly increased with MLN8237; The qRT-PCR results showed the expression of p16 and p21 was significantly up-regulated after IR with MLN8237( Aurora kinase A inhibitor);The results of Luciferase analysis showed that the activity of E2 F was significantly decreased while the activity of JNK was significantly increased by inhibition of Aurora kinase A( P < 0. 01). Conclusion:Alisertib can inhibit the proliferation and induce apoptosis of NCI-H1975 cells by targeting Aurora kinase A,thereby increasing the radiosensitivity of NCI-H1975 cells.

关键词(KeyWords): 癌,非小细胞肺;转染;辐射耐受性;细胞凋亡;细胞增殖;阿立塞替
carcinoma,non-small-cell lung(NSCLC);transfection;radiation tolerance;apoptosis;cell proliferation;alisertib

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31560270)

作者(Author): 李舜;杨智松;韩国征;李红艳;齐浩明;
LI Shun;YANG Zhisong;HAN Guozheng;LI Hongyan;QI Haoming;Daqing Longnan Hospital;Department of Traditional Chinese Medicine,the First Affiliated Hospital of Shihezi University;Department of Cardiovascular Medicine,Weinan Central Hospital;

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DOI: 10.19367/j.cnki.2096-8388.2020.05.006

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