汉防己甲素对人鼻咽癌CNE1和CNE2细胞株的放疗增敏作用

发布时间：2018-05-11 01:42

本文选题：汉防己甲素 + 鼻咽癌　；参考：《中国病理生理杂志》2017年09期

【摘要】：目的:探讨最大非毒性剂量汉防己甲素(tetrandrine,Tet)对人鼻咽癌细胞株CNE1和CNE2的放疗增敏机制。方法:分别采用最大非毒性剂量Tet(对CNE1细胞为1.5μmol/L;对CNE2细胞为1.8μmol/L)、4 Gy放疗和最大非毒性剂量Tet联合放疗处理CNE1和CNE2细胞;流式细胞术检测各组细胞周期分布,Western blot检测各组细胞γ-H2AX、cleaved caspase-3、p-CDC25C、CDK1、p-CDK1、cyclin B1、ERK和p-ERK的蛋白水平。结果:最大非毒性剂量Tet联合放疗后可上调CNE1和CNE2细胞中γ-H2AX的表达。放疗组CNE1和CNE2细胞G_2/M期的比例分别为(18.09±0.42)%和(18.48±1.32)%,联合处理组CNE1和CNE2细胞G_2/M期的比例降为(15.88±1.04)%和(13.80±0.82)%,与放疗组比较差异有统计学意义(P0.05)。联合处理可增加放疗所致的cleaved caspase-3的蛋白水平(P0.05)。不同浓度Tet处理CNE1和CNE2细胞后,p-CDC25C和p-CDK1的蛋白水平随Tet浓度增加而升高(P0.05),CDK1的表达无明显改变;最大非毒性剂量Tet不影响p-CDC25C、p-CDK1和CDK1的蛋白水平。在CNE1和CNE2细胞中,联合处理可明显降低放疗引起的p-CDC25C和p-CDK1的蛋白水平(P0.05),上调放疗后cyclin B1的表达,而对总CDK1的表达无明显调节作用;联合处理可显著抑制放疗所致的pERK蛋白水平(P0.05)。结论:最大非毒性剂量Tet可以增加放疗引起的CNE1和CNE2细胞的DNA断裂及细胞凋亡,其放疗增敏的机制可能与Tet调控ERK/CDC25C/CDK1/cyclin B1通路、去除放疗导致的G2/M期阻滞有关。
[Abstract]:Aim: to investigate the mechanism of radiosensitization of human nasopharyngeal carcinoma cell line CNE1 and CNE2 by maximum nontoxic dose Tetrandrine Tet. Methods: CNE1 and CNE2 cells were treated with maximum nontoxic dose of tett (1.5 渭 mol / L for CNE1 cells, 1.8 渭 mol / L of CNE2 cells with 4 Gy radiotherapy and maximum nontoxic dose of Tet combined with radiotherapy). Cell cycle distribution in each group was detected by flow cytometry. Western blot was used to detect the protein levels of 纬 -H2AXCleaved caspase-3pCDC25, CDK1, CDK1, p-CDK1, cyclin B1, ERK and p-ERK in each group. Results: the expression of 纬 -H2AX in CNE1 and CNE2 cells could be upregulated after maximum nontoxic dose of Tet combined with radiotherapy. The ratio of G 2 / M phase of CNE1 and CNE2 cells in radiotherapy group was 18.09 卤0.42% and 18.48 卤1.32%, respectively. The ratio of G 2 / M phase of CNE1 and CNE2 cells in combined treatment group was 15.88 卤1.04% and 13.80 卤0.82% respectively, which was significantly higher than that in radiotherapy group (P 0.05). Combined treatment could increase the protein level of cleaved caspase-3 induced by radiotherapy (P 0.05). The protein levels of p-CDC25C and p-CDK1 in CNE1 and CNE2 cells treated with different concentrations of Tet were increased with the increase of Tet concentration, and the expression of P0.05 and CDK1 was not significantly changed, while the maximum non-toxic dose of Tet did not affect the protein levels of p-CDC25Cp-CDK1 and CDK1. In CNE1 and CNE2 cells, combined treatment significantly decreased the protein level of p-CDC25C and p-CDK1 induced by radiotherapy and up-regulated the expression of cyclin B1, but did not regulate the expression of total CDK1, while combined treatment significantly inhibited the level of pERK protein (P0.05) induced by radiotherapy. Conclusion: the maximum nontoxic dose of Tet can increase DNA fragmentation and apoptosis of CNE1 and CNE2 cells induced by radiotherapy. The mechanism of radiosensitization may be related to Tet regulation of ERK/CDC25C/CDK1/cyclin B1 pathway and removal of G 2 / M phase arrest induced by radiotherapy.
【作者单位】：中山大学附属第三医院耳鼻咽喉头颈外科;
【基金】：广东省自然科学基金资助项目(No.2014A030313142) 广州市科技计划对外科技合作专项(No.2013J4500013) 广州市天河区科技计划项目医疗卫生专项(No.2013KW027)
【分类号】：R739.63

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