中华细胞与干细胞杂志（电子版）
中華細胞與榦細胞雜誌（電子版）
중화세포여간세포잡지（전자판）
CHINESE JOURNAL OF CELL AND STEM CELL
2013年
4期
191-196
,共6页
陈旭征%黎金浓%胡丹%曹治云%廖联明%杜建
陳旭徵%黎金濃%鬍丹%曹治雲%廖聯明%杜建
진욱정%려금농%호단%조치운%료련명%두건
肝肿瘤%干细胞%抗药性,肿瘤%基因
肝腫瘤%榦細胞%抗藥性,腫瘤%基因
간종류%간세포%항약성,종류%기인
Liver neoplasm%Stem cell%Drug resistance,neoplasm%Genes
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。
目的：探討解毒消癥飲乙痠乙酯提取物（EE-JXY)能否降低肝癌細胞株Huh7對小劑量氟尿嘧啶（5-FU）化療耐藥的影響，併從腫瘤榦細胞的角度探討可能的機製。方法EE-JXY聯閤小劑量5-FU體外榦預人肝癌細胞株Huh7。MTT法檢測細胞存活率；流式細胞儀分析Huh7肝癌細胞株中側群（side population，SP）細胞的比例；PCR檢測肝癌榦細胞相關基因ATP結閤盒轉運蛋白G2（ABCG2）和八聚體結閤轉錄因子4（Oct4）mRNA的錶達。採用單因素方差分析對數據進行統計。結果小劑量5-FU對Huh7細胞的增殖有抑製作用[24 h存活率（84.54±2.88）﹪，96 h存活率（58.36±3.52）﹪]，但SP細胞的比例也從24 h的4.72﹪升高至96 h的12.07﹪，同時誘導瞭ABCG2和Oct4 mRNA錶達的上調。EE-JXY能夠增彊小劑量5-FU對Huh7細胞增殖的抑製能力[24h存活率從（84.54±2.88）﹪下降到（31.23±2.42）﹪，96 h存活率從（58.36±3.52）﹪下降到（25.37±0.99）﹪]，SP含量從24 h的4.72﹪下降至1.69﹪，96 h的12.07﹪下降至9.97﹪，同時也抑製瞭ABCG2和Oct4 mRNA的錶達。結論肝癌Huh7細胞對小劑量5-FU存在化療牴抗。EE-JXY能通過降低肝癌榦細胞相關基因ABCG2 mRNA和Oct4 mRNA水平及SP的比例，增彊Huh7細胞對小劑量5-FU的化療敏感性，這可能是解毒消癥飲降低腫瘤複髮和轉移的機製之一。
목적：탐토해독소징음을산을지제취물（EE-JXY)능부강저간암세포주Huh7대소제량불뇨밀정（5-FU）화료내약적영향，병종종류간세포적각도탐토가능적궤제。방법EE-JXY연합소제량5-FU체외간예인간암세포주Huh7。MTT법검측세포존활솔；류식세포의분석Huh7간암세포주중측군（side population，SP）세포적비례；PCR검측간암간세포상관기인ATP결합합전운단백G2（ABCG2）화팔취체결합전록인자4（Oct4）mRNA적표체。채용단인소방차분석대수거진행통계。결과소제량5-FU대Huh7세포적증식유억제작용[24 h존활솔（84.54±2.88）﹪，96 h존활솔（58.36±3.52）﹪]，단SP세포적비례야종24 h적4.72﹪승고지96 h적12.07﹪，동시유도료ABCG2화Oct4 mRNA표체적상조。EE-JXY능구증강소제량5-FU대Huh7세포증식적억제능력[24h존활솔종（84.54±2.88）﹪하강도（31.23±2.42）﹪，96 h존활솔종（58.36±3.52）﹪하강도（25.37±0.99）﹪]，SP함량종24 h적4.72﹪하강지1.69﹪，96 h적12.07﹪하강지9.97﹪，동시야억제료ABCG2화Oct4 mRNA적표체。결론간암Huh7세포대소제량5-FU존재화료저항。EE-JXY능통과강저간암간세포상관기인ABCG2 mRNA화Oct4 mRNA수평급SP적비례，증강Huh7세포대소제량5-FU적화료민감성，저가능시해독소징음강저종류복발화전이적궤제지일。
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.
