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Genistein对受辐射L-02人肝细胞增殖和DNA损伤的影响
类春燕1,沈秀华2,张乃宁1,丛峰松3,马俐君4*,宋立华1,5*
0
(1. 上海交通大学农业与生物学院食品科学与工程系,上海200240
2. 上海交通大学医学院营养系,上海200025
3. 上海交通大学生命科学技术学院实验教学中心,上海200240
4. 上海交通大学医学院附属仁济医院长宁分院肿瘤科,上海200050
5. 上海交通大学陆伯勋食品安全研究中心,上海200240
*通信作者)
摘要:
目的 探讨金雀异黄素(genistein,GEN)对L-02人肝细胞DNA辐射损伤的防护作用。方法 (1)L-02细胞经不同浓度(1、5、10、20 μmol/L) GEN预处理24 h后,接受不同剂量(6、8、12、16、20 Gy) X线照射,利用MTT法检测细胞增殖情况。(2)L-02细胞经不同浓度GEN预处理后接受8 Gy (300 cGy/min) X线照射,利用单细胞凝胶电泳检测照射后不同时间(24、48、72 h)细胞DNA损伤情况。另设正常对照(N)、单纯GEN处理(G)及单纯照射(R)组作为参照。结果 (1)当照射剂量为6、8及12 Gy时,5 μmol/L GEN预处理组细胞增殖率显著高于R组(P<0.05),而照射剂量为16 Gy和20 Gy时,各浓度GEN预处理组细胞增殖率与R组相比未见明显升高。(2)DNA损伤检测结果显示,N组和不接受照射的各浓度G组细胞均未见彗星拖尾。经8 Gy X线照射后24 h,R组及各浓度GEN预处理组彗星出现率均小于1%,各组间彗星尾长差异无统计学意义;照后48 h, R组彗星出现率达(24.2±1.2)%,彗星尾长达(283.6±22.3) μm,而各浓度GEN预处理组的彗星出现率和彗星尾长均较R组明显降低(P<0.05);照后72 h,R组彗星出现率较48 h明显下降(P<0.05)、彗星尾长明显缩短(P<0.05),1、5 μmol/L GEN预处理组彗星出现率和彗星尾长仍明显低于R组(P<0.05),但10、20 μmol/L GEN预处理组彗星出现率则明显升高(P<0.05),彗星尾长明显增长(P<0.05)。 结论 低浓度(1、5 μmol/L) GEN能有效减轻较低剂量(8 Gy)照射对L-02人肝细胞的DNA辐射损伤。
关键词:  金雀异黄素  肝细胞  X线  辐射损伤  DNA损伤
DOI:10.3724/SP.J.1008.2012.00359
投稿时间:2011-10-31修订日期:2012-03-06
基金项目:国家自然科学基金(30972460),上海市自然科学基金(09ZR1415400).
Effects of genistein on proliferation and DNA damage of irradiated human liver L-02 cells
LEI Chun-yan1,SHEN Xiu-hua2,ZHANG Nai-ning1,CONG Feng-song3,MA Li-jun4*,SONG Li-hua1,5*
(1. Department of Food Science and Engineering, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai200240, China
2. Department of Nutrition, School of Medicine, Shanghai Jiaotong University, Shanghai200025, China
3. Experimental Teaching Center, School of Life Science and Technology, Shanghai Jiaotong University, Shanghai200240, China
4. Department of Oncology, Changning Branch, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200050, China
5. Bor S. Luh Food Safety Research Center, Shanghai Jiaotong University, Shanghai 200240, China
*Corresponding authors.)
Abstract:
Objective To investigate the radioprotective effects of genistein (GEN) against radiation-induced DNA damage in human liver cell line L-02.Methods (1)L-02 cells were treated with different concentrations of GEN (1, 5, 10, and 20 μmol/L) for 24 h, and then irradiated with X-ray at the doses of 6, 8, 12,16,and 20 Gy. Forty-eight hours after irradiation, MTT method was applied to examine the proliferation of L-02 cells.(2)L-02 cells were treated with different concentrations of GEN (1, 5, 10, and 20 μmol/L) for 24 h, and then irradiated with X-ray at the doses of 8 Gy (300 cGy/min) . Single cell gel electrophoresis was used to determine the DNA damage after radiation . Results (1)After irradiation with 6, 8 and 12 Gy of X-ray, the cell proliferation rate of 5 μmol/L GEN-pretreated group was significantly increased compared to radiation alone (R) group (P<0.05). But no significant increase was observed in GEN-pretreated groups irradiated with 16 Gy and 20 Gy of X-ray compared with R group. (2)As for the DNA damage, no comet cells were detected in normal control group or all GEN-treated groups without irradiation. After irradiation with 8 Gy of X-ray for 24 h, comet incidences were less than 1% in all GNE-pretreated groups and R group, and comet tail length showed no significant difference between different groups. At 48 h after irradiation, the comet incidence of R group was (24.2±1.2)% and the comet tail length was (283.6±22.3) μm, while both comet incidence and tail length of GEN-pretreated groups were significantly lower than those of R group (P<0.05). The comet incidence and tail length of R group were significantly decreased 72 h after irradiation compared with 48 h after irradiation (P<0.05), and those in 1 μmol/L and 5 μmol/L GEN-pretreated groups were still significantly lower than those of R group (P<0.05), but the comet incidence and tail length of 10 μmol/L and 20 μmol/L GEN-pretreated groups were significantly increased compared to those of R group (P<0.05). Conclusion Low concentration of GEN (1 and 5 μmol/L) can effectively protect the human liver cells L-02 against X-ray (8 Gy)-induced DNA damage.
Key words:  genistein  hepatocytes  X-rays  radiation injuries  DNA damage