학술논문
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
Document Type
Academic Journal
Author
梁慧玲; 何晓琴; 甘园园; 周宇杰; 熊琳; 唐甜; 徐细明; LIANG Huiling; HE Xiaoqin; GAN Yuanyuan; ZHOU Yujie; XIONG Lin; TANG Tian; XU Ximing
Source
中国医药导报 / China Medical Herald. 14(36):4-20
Subject
Language
Chinese
ISSN
1673-7210
Abstract
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 GyX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SHIP2mRNA相对表达量,Westem-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 GyX线照射处理的Hep-2细胞GJM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.
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.RTqPCR 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.
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.RTqPCR 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.