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作 者:申安[1] 王雪莲[1] 宁厚法[1] 马炳奇[1] 李世平[1]
机构地区:[1]潍坊医学院附属医院肝胆外科,山东261031
出 处:《中华肝胆外科杂志》2017年第2期130-133,共4页Chinese Journal of Hepatobiliary Surgery
摘 要:目的探讨细胞凋亡抑制蛋白1(cIAP-1)对肝癌SMMC-7721细胞放疗敏感性的影响。方法首先构建特异性沉默cIAP-1基因的pGCsi—H1-shRNA载体;然后通过CCK8试验、蛋白印迹检测(Western blot)、实时定量.PCR(qRT—PCR)、流式细胞仪和凋亡检测等方法分析照射处理后细胞活性、细胞周期以及细胞凋亡的变化。结果分别在不同照射剂量(1Gy、4Gy、7Gy、10Gy)条件下检测肝癌SMMC-7721细胞24h细胞增殖率。与对照组(pGCsi—H1-control)比较,沉默cIAP-1基因细胞组(pGCsi—H1-shRNA)在各个照射剂量条件下细胞增殖率均明显降低,并且存在照射剂量依赖性(P〈0.05)。与未照射处理组比较,照射处理组SMMC-7721细胞明显阻滞在G1/G0期(P〈0.01),处于S期细胞比例也明显降低(P〈0.01)。与对照组比较,沉默cIAP-1基因组细胞周期明显阻滞在G1/G0期(P〈0.05),沉默cIAP-1基因组细胞周期凋亡百分比显著增加(P〈0.05)。结论沉默cIAP-1表达可提高肝癌对放疗的敏感性,并能协同放疗抑制肝癌细胞的增殖率。Objective To explore the effect of cellular inhibitor of apoptosis proteinl ( cIAP-1 ) gene on the radiosensitivity of SMMC-7721 cells. Methods We silenced cIAP-1 expressions by the shRNA technology, and then we detected the changes of cell proliferation, cell cycle and cell apoptosis by CCK8 assay, Western blot, qRT-PCR and flow cytometry after the radiotherapy. Results The cell proliferation rate of liver cancer SMMC-7721 cells at different radiation doses of 1 Gy, 4 Gy, 7 Gy and 10 Gy was detected. Comparing with control group (pGCsi-H1-control), the cell proliferation in cIAP-1 silencing group (pGCsi- H1-shRNA) was significantly reduced at various radiation doses, and the effect was dose-dependent (P 〈 0. 05). G1/G0 phase arrest was observed after radiation (P 〈0.01 ), and the proportion of cells in S phase was significantly reduced compared with control (P 〈 0.01 ). Compared with control group, G1/G0 phase arrest was detected ( P 〈 0.05 ), and the percentage of cell apoptosis was increased significantly in cIAP-1 silencing group (P 〈 0.05 ). Conclusion cIAP-1 silencing can enhance the radiosensitivity of liver cancer cells, and inhibit cell proliferation by promoting the anti-tumor effect of radiation.
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