摘要
目的:研究10-羟基喜树碱对肝癌细胞Hep G2的DNA甲基化水平的调节作用。方法:体外培养Hep G2细胞,分为给药组(10-羟基喜树碱25μg/ml)和空白对照组,每组6个小组,每小组5个平行孔,分别于给药后24、48、72 h各组取2个小组,一组用MTT法检测Hep G2细胞的生长抑制率;另一组提取Hep G2细胞的DNA,酸水解后高效液相色谱法检测其甲基化率。结果:10-羟基喜树碱作用24、48、72 h后,Hep G2细胞的生长抑制率分别为(61.6±4.9)%、(85.7±0.7)%、(97.9±0.7)%;给药组Hep G2细胞DNA甲基化率分别为(2.81±0.34)%、(6.67±0.24)%、(6.83±0.24)%,明显高于相应的空白对照组[(1.88±0.13)%、(1.91±0.11)%、(1.98±0.18)%](P<0.05),且与作用时间呈正相关。结论:10-羟基喜树碱在抑制Hep G2细胞的生长的同时提高了细胞DNA的甲基化水平。
OBJECTIVE: To study the effects of 10-hydroxycamptothecine (10-HCPT) on the regulation of DNA methylation level in liver cancer cell HepG2. METHODS: Liver cancer cell HepG2 was cultured in vitro and divided into medication group (10-HCPT 25 μg/ml) and blank control group with 6 subgroups in each group. There were 5 parallel holes in each subgroup. 2 sub- groups were sampled in each group 24, 48 and 72 h after medication. For one subgroup, the growth inhibition rates of HepG2 was detected by using MTT assay; for another subgroup, DNA of HepG2 and the parallel test cell were extracted respectively, and the methylation rate was determined by HPLC after acid hydrolysis. RESULTS: After treated with 10-HCPT for 24, 48 and 72 h, the growth inhibition rates of HepG2 were (61.6 ± 4.9)%, (85.7 ± 0.7)% and (97.9 ± 0.7)%. The DNA methylation rate of medication group were (2.81 ± 0.34)%, (6.67 ± 0.24)% and (6.83 ± 0.24)%, which were significantly higher than those of blank control group [(1.88 ± 0.13% ), (1.91± 0.11)% and (1.98± 0.18)%] (P〈0.05), positively associated with treatment time. CONCLUSIONS: 10-HCPT may increase the overall DNA methylation level during inhibiting cancer cell.
出处
《中国药房》
CAS
CSCD
2014年第45期4245-4247,共3页
China Pharmacy
基金
湖南省教育厅科研项目(No.11C0943)
湖南省"十二五"中药学重点学科资助项目(No.湘教通〔2011〕76)
关键词
10-羟基喜树碱
甲基化
肝癌细胞Hep
G2
抑癌机制
10-hydroxycamptothecine
Methylation
Liver cancer cell HepG2
Cancer inhibition mechanism