SHIP2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  

Study on the relation of SHIP2 with proliferation,apoptosis and cell cycle in Hep-2 cells after radiation

作　　者：梁慧玲[1] 何晓琴[1] 甘园园[1] 周宇杰[1] 熊琳 唐甜[1] 徐细明[1] 

机构地区：[1]武汉大学人民医院肿瘤中心,湖北武汉430060

出　　处：《中国医药导报》2017年第36期4-8,20,共6页China Medical Herald

基　　金：国家自然科学基金资助项目青年项目(81402242)

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-q PCR检测SHIP2m RNA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m RNA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。Objective To explore the relation between the expression of SHIP2 and proliferation, apoptosis and cell cy- cle 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 gToup, Hep-2 ceils＂ 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 remark- able increase in apoptosis of the experiment group （all P 〈 0.01）. After treated with 4 Gy and 8 Gy X-ray G2/M phase ar- rest 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.

关 键 词：X线 SHIP2 增殖 凋亡 细胞周期 

分 类 号：R739.65[医药卫生—肿瘤]