HIP2在放射线处理后喉癌Hep-2细胞中的增殖、凋亡及细胞周期中作用

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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。

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	梁慧玲何晓琴甘园园周宇杰熊琳唐甜徐细明

关键词 ： X线, SHIP2, 增殖, 凋亡, 细胞周期

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 group was lower than those of the control group (all P ＜ 0.01). Conclusion In Hep-2 cells, low expression of SHIP2 after the treatment with X-ray, may be one important pathway of X-ray inhibiting cell proliferation and increasing cell apoptosis and G2/M phase arrest.

Key words： X-ray SHIP2 Proliferation Apoptosis Cell cycle

基金资助:国家自然科学基金资助项目青年项目（814022 42）。

通讯作者: 徐细明（1976-），男，博士，主任医师，副教授，硕士生导师；研究方向：肝癌等肿瘤的发病机制与防治。

作者简介: 梁慧玲(1991-)，女，武汉大学第一临床学院2015级肿瘤学专业在读硕士研究生；研究方向：头颈部肿瘤、消化道肿瘤的发病机制与防治。

引用本文:

梁慧玲何晓琴甘园园周宇杰熊琳唐甜徐细明. HIP2在放射线处理后喉癌Hep-2细胞中的增殖、凋亡及细胞周期中作用[J]. 中国医药导报, 2017, 14(36): 4-8,20.
LIANG Huiling HE Xiaoqin GAN Yuanyuan ZHOU Yujie XIONG Lin TANG Tian XU Ximing. Study on the relation of SHIP2 with proliferation, apoptosis and cell cycle in Hep-2 cells after radiation. 中国医药导报, 2017, 14(36): 4-8,20.

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http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2017/V14/I36/4

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