中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
10期
764-767
,共4页
祝葆华%蒲荣%张国平%李明意%王兰天%苑进凯
祝葆華%蒲榮%張國平%李明意%王蘭天%苑進凱
축보화%포영%장국평%리명의%왕란천%원진개
癌,肝细胞%恶性表型%柴胡皂甙-d%p27
癌,肝細胞%噁性錶型%柴鬍皂甙-d%p27
암,간세포%악성표형%시호조대-d%p27
Carcinoma,hepatocellular%Malignant phenotype%Saikosaponins-d%p27
目的 探讨柴胡皂甙-d (SSd)对人肝癌细胞株HepG2细胞恶性表型的逆转作用及其机制. 方法 常规培养HepG2细胞至对数生长期,四甲基偶氮唑盐(MTT)比色法观察SSd不同浓度、作用不同时间对HepG2细胞增殖情况的影响.实验组选择10 mg/L的SSd作用HepG2细胞48h,Giemsa染色法观察细胞形态学改变;化学发光法测定细胞上清液中甲胎蛋白(AFP)浓度;放射免疫法检测细胞上清液中白蛋白浓度;transwell小室实验观察细胞迁移率;RT-PCR检测p27基因mRNA表达.并与空白对照组做比较.数据在多组间比较采用随机分组单因素方差分析,在两组间比较采用t检验. 结果 10 mg/L的SSd作用48 h,对HepG2细胞的生长抑制最明显(F=265.06,P＜0.01).实验组HepG2细胞形态倾向于正常分化,形态变小、变圆,核变小、变圆或卵圆形,核/质比例缩小.与对照组比较,实验组穿膜细胞数明显减少[(50.60+4.04)个与(41.00+4.64)个,t= -7.32,P＜0.01];细胞上清液中白蛋白含量明显增多[(24.7+2.8)×10-3g/L与(31.1 +4.9)×10-3g/L,t=7.83,P＜ 0.05];AFP含量明显减少[(118.2±15.6)×10μ g/L与(70.3+5.7)×103μg/L,t=-10.72,P＜0.0l].实验组p27基因mRNA的相对表达量明显多于对照组(t=22.00,P＜0.05). 结论 SSd对人肝癌细胞株HepG2细胞恶性表型具有明显的逆转作用,其机制可能与SSd上调HepG2细胞p27基因mRNA表达使其进入分化过程有关.
目的 探討柴鬍皂甙-d (SSd)對人肝癌細胞株HepG2細胞噁性錶型的逆轉作用及其機製. 方法 常規培養HepG2細胞至對數生長期,四甲基偶氮唑鹽(MTT)比色法觀察SSd不同濃度、作用不同時間對HepG2細胞增殖情況的影響.實驗組選擇10 mg/L的SSd作用HepG2細胞48h,Giemsa染色法觀察細胞形態學改變;化學髮光法測定細胞上清液中甲胎蛋白(AFP)濃度;放射免疫法檢測細胞上清液中白蛋白濃度;transwell小室實驗觀察細胞遷移率;RT-PCR檢測p27基因mRNA錶達.併與空白對照組做比較.數據在多組間比較採用隨機分組單因素方差分析,在兩組間比較採用t檢驗. 結果 10 mg/L的SSd作用48 h,對HepG2細胞的生長抑製最明顯(F=265.06,P＜0.01).實驗組HepG2細胞形態傾嚮于正常分化,形態變小、變圓,覈變小、變圓或卵圓形,覈/質比例縮小.與對照組比較,實驗組穿膜細胞數明顯減少[(50.60+4.04)箇與(41.00+4.64)箇,t= -7.32,P＜0.01];細胞上清液中白蛋白含量明顯增多[(24.7+2.8)×10-3g/L與(31.1 +4.9)×10-3g/L,t=7.83,P＜ 0.05];AFP含量明顯減少[(118.2±15.6)×10μ g/L與(70.3+5.7)×103μg/L,t=-10.72,P＜0.0l].實驗組p27基因mRNA的相對錶達量明顯多于對照組(t=22.00,P＜0.05). 結論 SSd對人肝癌細胞株HepG2細胞噁性錶型具有明顯的逆轉作用,其機製可能與SSd上調HepG2細胞p27基因mRNA錶達使其進入分化過程有關.
목적 탐토시호조대-d (SSd)대인간암세포주HepG2세포악성표형적역전작용급기궤제. 방법 상규배양HepG2세포지대수생장기,사갑기우담서염(MTT)비색법관찰SSd불동농도、작용불동시간대HepG2세포증식정황적영향.실험조선택10 mg/L적SSd작용HepG2세포48h,Giemsa염색법관찰세포형태학개변;화학발광법측정세포상청액중갑태단백(AFP)농도;방사면역법검측세포상청액중백단백농도;transwell소실실험관찰세포천이솔;RT-PCR검측p27기인mRNA표체.병여공백대조조주비교.수거재다조간비교채용수궤분조단인소방차분석,재량조간비교채용t검험. 결과 10 mg/L적SSd작용48 h,대HepG2세포적생장억제최명현(F=265.06,P＜0.01).실험조HepG2세포형태경향우정상분화,형태변소、변원,핵변소、변원혹란원형,핵/질비례축소.여대조조비교,실험조천막세포수명현감소[(50.60+4.04)개여(41.00+4.64)개,t= -7.32,P＜0.01];세포상청액중백단백함량명현증다[(24.7+2.8)×10-3g/L여(31.1 +4.9)×10-3g/L,t=7.83,P＜ 0.05];AFP함량명현감소[(118.2±15.6)×10μ g/L여(70.3+5.7)×103μg/L,t=-10.72,P＜0.0l].실험조p27기인mRNA적상대표체량명현다우대조조(t=22.00,P＜0.05). 결론 SSd대인간암세포주HepG2세포악성표형구유명현적역전작용,기궤제가능여SSd상조HepG2세포p27기인mRNA표체사기진입분화과정유관.
Objective To observe the effect of SSd on reversing the malignant phenotype of HepG2cells and to investigate its mechanism in order to prove that SSd is a new choice to prevent and treat HCC.Methods HepG2 cells were cultured and treated by different concentrationgs (0 mg/L,2.5 mg/L,5.0 mg/L,10.0 mg/L and 20.0 mg/L) of SSd for 24 h,and treated by 10 mg/L of SSd for 0 h,6 h,12 h,24 h,48 h and 72h respectively.The cell inhibition rates were measured by MTT assay.Then cells were treated by 10 mg/L SSd for 48 hr in experimental group and treated by no SSd as a control,their morphological changes were observed by contrast phase microscope.The concentrations of ALB and AFP in clear supernatant liquid of cells were detected by radio-immunity and chemiluminescence.The cell migration rates were observed by transwell method,the relative expression levels of p27 mRNA were messured by RT-PCR.Results The inhibitive effect of 10 mg/L SSd was the most significant among different concentrations (F=265.06,P＜0.01).The shape of HepG2from experimental group turned into small and round,and their volume ratios of nucleus to plasma decreased.ALB in supernatant liquid of HepG2 was higher (t= 7.83,P ＜ 0.05,and its AFP was lower (t=-10.72,P ＜ 0.01 )as compared to control group.Cells migrated were fewer and p27 mRNA expression of HepG2 was higher in experimental group than that in control group (t=22.00,P＜0.05).Conclusion SSd could reverse the malignant phenotype of HepG2 cells.It was suggested that the up-regulation of p27 mRNA expression play an important role in the differentiation of HepG2 cells treated by SSd.
