绿茶主要活性成分对叙利亚地鼠胚胎癌前细胞生长和凋亡的影响  

Effects of cell growth and apoptosis of preneoplastic Syrian hamster embryo cells by green tea constituent epigallocatechin-3-gallate

作　　者：焦振泉[1] 吕相征[1] 郭云昌[1] 阚飚[2] 徐勇[3] 

机构地区：[1]中国疾病预防控制中心营养与食品安全所,北京100050 [2]中国疾病预防控制中心传染病预防控制所,北京100050 [3]美国印第安纳大学药理和毒理系

出　　处：《中华预防医学杂志》2007年第5期380-386,共7页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金(30400357)

摘　　要：目的建立叙利亚地鼠胚胎细胞(Syrian hamster embHo cell,SHE 细胞)混合培养模型,研究表没食子儿茶素没食子酸酯(epigallocatechin-3-Gallate,EGCG)对 SHE 癌前细胞生长、增殖和凋亡的作用,探讨 EGCG 的抑癌机制。方法将 SHE 癌前细胞和正常细胞分别按1:10 000、1:1000、1:100、1:10比例接种于6孔的培养皿中,培养7d,建立 SHE 细胞混合培养模型。以0μmol/L EGCG为对照组,分别选取浓度为0.5、1、5、10、50 μmol/L 的 EGCG,通过 SHE 细胞生长实验,原位细胞凋亡实验、原位细胞增殖实验和基因芯片检测 EGCG 对 SHE 正常细胞、SHE 癌前细胞和混合培养模型中SHE 癌前细胞的生长、增殖、凋亡及相关调控基因表达的影响。结果 SHE 癌前细胞与正常细胞的比例为1:100是合适的混合培养模型。0.5、1、5、10μmol/L 的 EGCG 促进了 SHE 正常细胞的生长,而浓度为50 μmol/L 的 EGCG 抑制了 SHE 正常细胞的生长。在1:200比例的 SHE 癌前细胞和正常细胞混合培养模型中,与对照组相比,浓度为5、10μmol/L 的 EGCG 抑制了不同大小的 SHE 癌前细胞克隆的生长。对照组中 SHE 癌前细胞的 DNA 合成指数和凋亡指数分别是39.3%和6.5%,5、10μmol/L EGCG 作用后 SHE 癌前细胞的 DNA 合成指数分别降至25.6%和24.8%,细胞凋亡指数分别升至12.65%和14.5%。EGCG 抑制了混合培养模型中 SHE 癌前细胞的生长和增殖,促进了其凋亡。EGCG 对于细胞凋亡的调控可能通过 p53、NF-κB、bel-2通道的基因表达来实现;EGCG 对于细胞增殖的调控可能通过阻滞细胞周期 G_1/S 期实现。结论 EGCG 可能是通过抑制癌前细胞的增殖和促进其凋亡进而抑制癌症。Objective The co-culture model of Syrian hamster embryo （SHE） normal （ primary cell） and preneoplastic cells mimicking in vivo status was establsihed and used to study the chemopreventive effects of epigallocatechin-3-Gallate （EGCG） on cell growth , proliferation , apoptosis and regulated genes expression of SHE preneoplastic cells and discussed on the mechanism of EGCG＇ s chemopreventive effect of carcinogeneisis. Methods The SHE cell preneoplastic and normal cells were cultured on the plates with 1：10 000,1：1000,1 ： 100, 1 ： 10 rates for 7days, and the co-culture model was established. The different concentration of EGCG （0,0. 5,1,5,10,50 μmol/L ）were used to treat the cells and the SHE cells growth assay, in situ cell apoptosis assay , in situ cell proliferation assay and microarray assay were used to determined the growth , apoptosis and proliferation of SHE preneoplastic cells . Results The co-culture model of SHE cells with the 1：100 rate between SHE preneoplastic cells and normal cells was established. 0. 5,1,5,10 μmol/L EGCG increased the colony growth and proliferation of SHE normal cells. In the coculture model of SHE cells with 1 ： 200 rate , compared the the control group, 5,10 μmol/L 的 EGCG suppressed the growthof different size of SHE preneoplastic cells clone. The DNA proliferation index and apoptosis index in the control group were 39. 3% - 6. 5% ,respectively. After treatment of 5,10 μmol/L EGCG , the proliferation index were decreased to 25.6% and 24. 8%, and the apoptosis index were increased to 12. 65% and 14. 5%. EGCG suppressed the growth and proliferation of SHE preneoplastic cells in co-culture model and increased its apoptosis. The pathway of cell apoptosis was regulated through the P53,NF-κB,bcl-2 signal pathway, and the pathway of cell proliferation was regulated through the growth arrest at G1/S phase of cell cycle. Conclusion The selective regulation of EGCG to normal and preneoplastic cells, the interaction of EGCG, SHE normal cells and SHE preneo

关 键 词：儿茶素 癌前状态 脱噬作用 

分 类 号：R285.5[医药卫生—中药学] R73-36[医药卫生—中医学]