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沉默APE1对非小细胞肺癌细胞的放射敏感性的影响研究

Silencing Effect of APE1 on the Radiosensitivity of Non-small Cell Lung Cancer Cells

作者:欧俞宏, 李轶, 刘琴等

Author:OU Yu-hong, LI Yi, LIU Qin.et al

收稿日期:          年卷(期)页码:2019,50(1):1-6

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:APE1 非小细胞肺癌 辐照 克隆形成 增殖 凋亡

Key words:APE1 Non-small lung cancer Radiation Colony formation Proliferation Apoptosis

基金项目:

中文摘要

目的 研究脱嘌呤脱嘧啶核酸内切酶-1(APE1)对非小细胞肺癌细胞放射敏感性的影响。方法 采用免疫组化染色检测手术标本切片(非小细胞肺癌组织20例及癌旁组织5例)中APE1的表达,Western blot与实时荧光定量(qRT)-PCR检测非小细胞肺癌细胞系(A549,H460,H1299)及肺正常上皮细胞系中APE1的表达;Western blot检测辐照A549与H460细胞0~6 Gy后APE1的表达;构建沉默APE1的A549与H460稳定细胞株后,Western blot检测其辐照6 Gy后APE1的蛋白表达;细胞克隆形成实验检测细胞克隆形成率;CCK-8检测细胞增殖;细胞术流式检测细胞凋亡。结果 非小细胞肺癌组织及细胞系中APE1的表达明显上调;辐照能诱导非小细胞肺癌细胞APE1的表达,且表达随辐照剂量增高而上调;沉默APE1能有效地抑制辐照诱导A549与H460细胞中APE1的表达;成功构建沉默APE1的A459、H460稳定细胞株;沉默APE1联合辐照进一步抑制了非小细胞肺癌细胞克隆形成率、细胞增殖率,同时促进细胞凋亡( PAPE1可增强非小细胞肺癌细胞的放射敏感性。

英文摘要

Objective To determine the effect of apurinic/apyrimidinic endonuclease-1(APE1) on the radiosensitivity of non-small cell lung cancer cells. Methods The expression of APE1 was detected by immunohistochemistry in 20 surgical specimen of non-small cell lung cancer tissues and 5 cases of pericarcinous lung tissues.The expressions of APE1 protein and mRNA in non-small cell lung cancer cell lines(A549,H460,H1299)and normal lung epithelial cell lines was detected by Western blot and qRT-PCR. The A549 and H460 stable cell lines with silencing of APE1 were constructed.The expression of APE1 in the silenced and non-silenced cells after exposure to 0-6 Gy radiation was detected by Western blot. The effect of silencing of APE1 was measured by colony formation assay,the proliferation of non-small cell lung cancer cells detected by CCK-8 assay, and the apoptosis of non-small cell lung cancer cells detected by Annexin Ⅴ/PI flow cytometry.Results The expression of APE1 was upregulated in non-small cell lung cancer tissues and cell lines.Radiation induced the expression of APE1 in non-small cell lung cancer cells in a dose responsive way. The silencing of APE1 inhibited the expression of APE1 induced by radiation in A549 and H460 cells. Exposure to radiation of the cells with silenced APE1 further inhibited cell colony formation and cell proliferation, and promoted the apoptosis of non-small cell lung cancer cells ( P<0.05). Conclusion Silencing of APE1 enhances the radiosensitivity of non-small cell lung cancer cells.

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