SHIP2在放射线处理后喉癌Hep―2细胞中增殖凋亡及细胞周期中作用.docVIP

SHIP2在放射线处理后喉癌Hep―2细胞中增殖凋亡及细胞周期中作用 　　[摘要] 目的 研究放疗后SHIP2在喉癌Hep-2细胞中的表达情况及与Hep-2细胞增殖、凋亡及细胞周期之间的对应关系。 方法 取对数生长期的喉癌Hep-2细胞，分为实验组（2、4、8 Gy X线）与对照组（0 Gy X线）两组，各组给予不同剂量X线处理后12、24、36、48 h，运用CCK-8检测Hep-2细胞增殖活力。根据细胞活力检测结果，确定射线处理后最适宜的时间点作为后续实验检测的具体时间点。实验组与对照组给予对应处理后特定时间，运用Annexin V-FITC/PI双染流氏细胞术检测细胞凋亡，PI单染流式细胞术检测细胞周期，RT-qPCR检测SHIP2 mRNA相对表达量，Western-blot检测SHIP2蛋白的相对表达量。 结果 与对照组比较，实验组Hep-2细胞活性降低（P 0.05），且随放射剂量递增，细胞增殖活力降低，处理后48 h检测结果最为显著。实验组各组Hep-2细胞凋亡数较对照组均明显增加（均P 0.01）。实验组中给予4 Gy、8 Gy X线照射处理的Hep-2细胞G2/M期阻滞与对照组相比显著增加（均P 0.01）。实验组Hep-2细胞中SHIP2的mRNA及蛋白相对表达量较对照组均有所降低（均P 0.01）。 结论 放射线处理后SHIP2的下调表达可能是X线抑制Hep-2细胞增殖、促进其细胞凋亡和G2/M期阻滞的重要途径。 　　[关键词] X线；SHIP2；增殖；凋亡；细胞周期 　　[中图分类号] R739.65 [文献标识码] A [文章编号] 1673-7210（2017）12（c）-0004-06 　　[Abstract] Objective To explore the relation between the expression of SHIP2 and proliferation， apoptosis and cell cycle in Hep-2 cells after radiotherapy. Methods Lupine carcinoma Hep-2 cells were divided into the experiment group （treated with 2， 4， 8 Gy X- ray） and the control group （treated with 0 Gy）. CCK-8 was repeatedly used to detect the proliferation of the cells 12， 24， 36 h and 48 h after the treatment. The most appropriate and exact time to detect cells in the subsequent experiment was set up following the result of proliferation. Then apoptosis was evaluated by AnnexinV- FITC/PI double staining flow cytometry and cell cycle was evaluated by PI single staining flow cytometry. RT-qPCR and Western-blot were used to detect the expression of SHIP2 mRNA and its protein. Results Compared with the control group， Hep-2 cells proliferation in the experiment group was inhibited （all P 0.05） . As the dose of ray increased， the viability descended， and the most remarkable result arose 48 h after the treatment. There was a remarkable increase in apoptosis of the experiment group （all P 0.01）. After treated with 4 Gy and 8 Gy X-ray G2/M phase arrest was remarkable too （all P 0.01）. The expression of SHIP2 mRNA and SHIP2 protein in the experiment gr