解毒消癥饮逆转氟尿嘧啶诱导的肝癌干细胞耐药的机制研究  

Jiedu xiaozheng yin reverses the resistance of human hepatoma Huh7 cell line to low-dose 5-FU

作　　者：陈旭征[1] 黎金浓[2] 胡丹[2] 曹治云[1] 廖联明[1] 杜建[1] 

机构地区：[1]福建中医药大学中西医结合研究院肿瘤研究所,福州350122 [2]福建中医药大学中西医结合研究院药学院,福州350122

出　　处：《中华细胞与干细胞杂志（电子版）》2013年第4期15-19,共5页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(81102582)

摘　　要：目的探讨解毒消癥饮乙酸乙酯提取物(EE-JXY)能否降低肝癌细胞株Huh7对小剂量氟尿嘧啶(5-FU)化疗耐药的影响,并从肿瘤干细胞的角度探讨可能的机制。方法EE-JXY联合小剂量5-FU体外干预人肝癌细胞株Huh7。MTT法检测细胞存活率;流式细胞仪分析Huh7肝癌细胞株中侧群(side population,SP)细胞的比例;PCR检测肝癌干细胞相关基因ATP结合盒转运蛋白G2(ABCG2)和八聚体结合转录因子4(Oct4)mRNA的表达。采用单因素方差分析对数据进行统计。结果小剂量5-FU对Huh7细胞的增殖有抑制作用[24 h存活率(84.54±2.88)﹪,96 h存活率(58.36±3.52)﹪],但SP细胞的比例也从24 h的4.72﹪升高至96 h的12.07﹪,同时诱导了ABCG2和Oct4 mRNA表达的上调。EE-JXY能够增强小剂量5-FU对Huh7细胞增殖的抑制能力[24h存活率从(84.54±2.88)﹪下降到(31.23±2.42)﹪,96 h存活率从(58.36±3.52)﹪下降到(25.37±0.99)﹪],SP含量从24 h的4.72﹪下降至1.69﹪,96 h的12.07﹪下降至9.97﹪,同时也抑制了ABCG2和Oct4 mRNA的表达。结论肝癌Huh7细胞对小剂量5-FU存在化疗抵抗。EE-JXY能通过降低肝癌干细胞相关基因ABCG2 mRNA和Oct4 mRNA水平及SP的比例,增强Huh7细胞对小剂量5-FU的化疗敏感性,这可能是解毒消癥饮降低肿瘤复发和转移的机制之一。Objective To investigate the effect of ethyl acetate extraction from Jiedu Xiaozheng Yin （EE-JXY） on resistance to 5-fluorouracil （5-FU） chemotherapy in Huh7 cell line, and explore the underlying mechanism from the aspect of cancer stem cells. Methods Huh7 cells were cultured in vitro with low-dose 5-FU in the present or absence of EE-JXY. The cell viability was measured using MTT method. The percentage of side population （SP） cells was analyzed by a flow cytometer. The mRNA levels of ABCG2 and Oct4 were examined using reverse transcription PCR. Results Low-dose 5-FU inhibited the proliferation of Huh7 cells, whereas increased the percentage of SP cells and up-regulated the expression of ABCG2 and Oct4 mRNA. EE-JXY not only enhanced the antitumor efficacy of low-dose 5-FU, but also counteracted low-dose 5-FU-induced increase of SP cells and levels of ABCG2 and Oct4 mRNA. Conclusion EE-JXY could reverse the resistance of Huh7 cells to low-dose 5-FU, the underlying mechanism may be related to inhibiting the ABCG2 and Oct4 mRNA expression and decreasing the proportion of SP cells.

关 键 词：肝肿瘤 干细胞 抗药性 肿瘤 基因 

分 类 号：R735.7[医药卫生—肿瘤]