摘要
目的探讨齐墩果酸对鼻咽癌CNE2细胞放疗敏感性的影响及其可能的机制,为提高鼻咽癌治疗效果提供理论依据。方法体外培养鼻咽癌CNE2细胞,分为4组:空白对照组(未处理)、齐墩果酸低剂量(2.5μmol/L齐墩果酸溶液)组、齐墩果酸中剂量(5.0μmol/L齐墩果酸溶液)组、齐墩果酸高剂量(10.0μmol/L齐墩果酸溶液)组。以6 MV-X线照射细胞,采用MTT实验检测细胞增殖情况并筛选最佳辐射剂量;采用流式细胞仪检测细胞凋亡情况;采用免疫印迹法检测凋亡相关蛋白[Bcl-2相关蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)、裂解的半胱氨酸天冬氨酸蛋白酶-3(caspase-3)]及通路相关蛋白[细胞命运决定子(Numb)、Notch1、SPLIT多毛增强子1重组蛋白(Hes1)、发状分裂相关增强子-5(Hes5)]表达水平。结果随着辐射剂量的增加(2、4、6、8 Gy)CNE2细胞增殖抑制率逐渐升高;同一辐射条件下,与空白对照组比较,齐墩果酸低、中、高剂量组CNE2细胞增殖抑制率均升高,差异有统计学意义(P<0.05),且呈剂量依赖性。照射剂量为4 Gy时CNE2细胞增殖抑制率为50%左右,因此选用照射剂量为4Gy进行后续实验。与空白对照组比较,齐墩果酸低、中、高剂量组CNE2细胞凋亡率均升高,差异有统计学意义(P<0.05);且呈剂量依赖性。与空白对照组比较,齐墩果酸低、中、高剂量组CNE2细胞Bcl-2、Notch1、Hes1、Hes5蛋白表达显著降低,Bax、caspase-3、Numb蛋白表达显著升高,差异均有统计学意义(P<0.05);且均呈剂量依赖性。结论齐墩果酸可抑制CNE2细胞增殖并促进其凋亡,增加其放疗敏感性,其作用机制可能与其促进Numb蛋白释放,抑制Notch信号通路活化有关。
Objective To investigate the effect of oleanolic acid on the radiosensitivity of nasopharyngeal carcinoma CNE2 cells and its possible mechanism,and provide theoretical basis for improving the therapeutic effect of nasopharyngeal carcinoma. Methods Nasopharyngeal carcinoma CNE2 cells were cultured in vitro and divided into four groups:blank control group(untreated),low dose oleanolic acid(2.5 μmol/L)group,medium dose oleanolic acid(5.0 μmol/L)group and high dose oleanolic acid(10.0 μmol/L)group. Cells were irradiated with 6 MV-X rays;MTT was used to detect cell proliferation and select the optimal radiation dose. Cell apoptosis was detected by Flow cytometry. The expression levels of apoptosis-related protein(Bax),B-cell lymphoma-2(Bcl-2),lytic caspase-3(caspase-3)and pathway-related proteins[cell fate determinants(Numb),Notch1,hairy and enhancer-of-split homologues 1(Hes1),hairy and enhancer of split 5(Hes5)]were detected by Western blotting. Results With the increase of radiation dose(2,4,6,8 Gy),the proliferation inhibition rate of CNE2 cells increased gradually;under the same radiation condition,compared with that in the blank control group,the proliferation inhibition rate of CNE2 cells in the low,medium and high dose oleanolic acid groups of increased(P<0.05)in a dose-dependent manner. Inhibition of CNE2 cell proliferation was about 50% at an irradiation dose of 4 Gy, so an irradiation dose of 4 Gy was chosen for subsequent experiments. Under the irradiation of 4 Gy,compared with the blank control group,the apoptosis rate of CNE2 cells in the low,medium and high dose oleanolic acid groups increased(P<0.05)in a dose-dependent manner. Compared with that in the blank control group,the protein expression levels of Bcl-2,Notch1,Hes1,Hes5 in the low,medium and high dose oleanolic acid groups were significantly lower(P<0.05),and the protein expression levels of Bax,caspase-3 and Numb were significantly higher(P<0.05) in a dose-dependent manner.Conclusions Oleanolic acid could inhibit the proliferation and increa
作者
曲丹菊
卢晓旭
韩振坤
莫颖禧
QU Dan-ju;LU Xiao-xu;HAN Zhen-kun;MO Ying-xi(Department of Otolaryngology,the First Affiliated Hospital of Henan University of Science&Technology,Luoyang,Henan 471000;Medical Insurance Office,Henan Cancer Hospital,Zhengzhou,Henan 450000;Department of Radiotherapy,the First Affiliated Hospital of Henan University of Science&Technology,Luoyang,Henan 471000;Guangxi Medical University,Nanning,Guangxi 530021,China)
出处
《热带医学杂志》
CAS
2021年第6期673-677,682,共6页
Journal of Tropical Medicine
基金
国家自然科学基金地区科学基金项目(81660447)。
