细胞周期相关基因微阵列在中药抑制肝癌细胞增殖作用机理研究中的应用

Investigation on the molecular mechanisms of anti-hepatocar cinoma herbs of traditional Chinese medicine by cell cycle microarray

目的 :设计DNA微阵列并利用其研究中药抗肿瘤的分子机制。方法 :制备了包含有与细胞周期和DNA损伤调控检测相关的 2 4个基因的cDNA微阵列 ,选择了本实验室已证明对肝癌细胞有抑制效果的中药 ,通过流式细胞仪检测其对细胞周期的影响 ,提取药物作用后的RNA ,反转录后标记探针与微阵列杂交 ,检测中药作用细胞后对细胞周期及损伤检测相关基因转录的影响。结果 :4种中药作用SMMC 772 1后 ,细胞周期基因和损伤检测点基因均有不同程度改变 ,表现为部分上调 ,部分下调 ,这些和流式细胞术检测的结果是相符合的。结论

Objective: To design DNA microarray and investigate the molecular anti-tumor mechanism of herbs of traditional chinese medicine. Method: cDNA microarrays consisting of 56 probes representing 24 human cell cycle genes were constructed, Four anti-hepatocarcinoma herbs including Radix Linderae,Hebra Artemisiae Annuae,Radix Arnebiae,Radix Astragli,were chosen. Effects of herbs on SMMC-7721 cell cycle were observed by flow cytometry assay. Effects of herbs on cell cycle gene expression in SMMC-7721 cells were analyzed by comparing hybridization of Dig-Labeled cDNAs from herb-treated cells and cDNAs from untreated cells. Result: Expressions of cell cycle genes were changed in different degrees after herbs treated. Some genes were down-regulated and some genes were up-regulated. The changes in gene expression agreed with the results of flow cytometry assay. Conclusion: The results suggest that these herbs may have effects on cell cycle and DNA damage checkpoint genes which may be the mechanism of the herbs,and DNA microarray can be used to investigate the biological function of extracts of traditional Chinese medicine.