[1]方姝煜,崔洁洁,龚梦嘉,等.全反式维甲酸诱导肝癌细胞Hepa1-6成熟和分化过程中的自噬[J].南方医科大学学报,2018,(05):527.
点击复制

全反式维甲酸诱导肝癌细胞Hepa1-6成熟和分化过程中的自噬()
分享到:

《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年05期
页码:
527
栏目:
出版日期:
2018-05-15

文章信息/Info

Title:
Changes in autophagy during maturation and differentiation of Hepa1-6 cells induced by all-trans retinoic acid
作者:
方姝煜崔洁洁龚梦嘉何昀张敬芳毕杨
关键词:
原发性肝癌全反式维甲酸自噬细胞分化
Keywords:
primary hepatic carcinoma all-trans retinoic acid autophagy cell differentiation
摘要:
目的探讨不同浓度全反式维甲酸(ATRA)对小鼠肝癌细胞Hepa1-6成熟分化及细胞自噬水平的影响。方法以小鼠肝癌 细胞Hepa1-6为研究对象,分别给予0.1、1、10 μmol/L浓度的ATRA处理,荧光定量PCR和Western blot检测肝细胞相关标志基 因的表达,吲哚菁绿(ICG)及过碘酸-希夫(PAS)染色检测小鼠肝癌细胞Hepa1-6的成熟功能,透射电镜观测细胞连接及自噬小 体,选择最适的ATRA作用浓度;Western blot检测自噬相关标志蛋白的表达,ptfLC3质粒转染细胞后共聚焦观察自噬流变化, 检测自噬流是否通畅。结果相较于空白对照组,ATRA可呈浓度依赖性抑制肝前体标志蛋白AFP的表达,并促进肝细胞成熟 标志ALB、CK18、TAT和ApoB的表达,ICG及PAS染色提示阳性细胞数明显增多;透射电镜结果显示ATRA诱导组的紧密连接 和细胞骨架明显增多,可见高尔基复合体以及较多的自噬小体和自噬溶酶体。均以10 μmol/L组最为显著,作为最适终浓度。 自噬相关标志蛋白LC3-II、Beclin-1、RAB7、P62的表达不同程度增高,LC3-Ⅱ/LC3-Ⅰ的比值明显增加(P<0.05),激光共聚焦可 见10 μmol/LATRA处理组细胞胞浆内绿色亮点的自噬小体及红色亮点的自噬溶酶体均较对照组明显增多。结论ATRA可诱 导小鼠肝癌细胞Hepa1-6的成熟分化,并促进细胞自噬水平增高。
Abstract:
Objective To investigate the effects of different concentrations of all-trans retinoic acid (ATRA) on the maturation, differentiation and autophagy of Hepa1-6 cells. Method Hepa1-6 cells were treated with 0.1, 1, and 10 μmol/L ATRA, and the changes in the expressions of hepatic specific markers were detected using real-time PCR and Western blotting. Indocyanine green (ICG) and periodic acid-schiff (PAS) staining was used to assess the functional maturation of Hepa1-6 cells, and the cellcell junction and autophagy were observed under transmission electron microscopy to determine the optimal concentration of ATRA for treatment. The expressions of autophagy- related markers in the cells were detected using Western blotting, and confocal microscopy was used to observe the autophagic flow in the cells transfected with ptfLC3 plasmid. Results Compared with the control cells, the hepatocytes treated with ATRA showed a concentration-dependent decrease in AFP expression and increase in the expressions of ALB, CK18, TAT and ApoB. ICG and PAS staining revealed significantly increased number of positive cells after ATRA treatment. Following ATRA treatment, the cells exhibited obviously increased tight junctions, cytoskeleton and number of autophagosomes under transmission electron microscopy. ATRA treatment resulted in significantly increased the expressions of autophagy- related markers LC3- II, Beclin- 1, RAB7 and P62 and also an increased ratio of LC3-Ⅱ/LC3-Ⅰ (P<0.05). Confocal microscopy revealed obviously increased green and red spots in the cells after ATRA treatment. Conclusion ATRA can induce the maturation and differentiation and enhance the level of autophagy in Hepa1-6 cells.

相似文献/References:

[1]陈丽,石燕,蒋成英,等.瘦素和瘦素受体在肝癌中的表达与临床意义[J].南方医科大学学报,2011,(05):830.
[2]黄达仁,左强,蔡红兵,等.组织多肽特异性抗原在原发性肝癌中的临床应用[J].南方医科大学学报,2005,(01):89.
 HUANG Da-ren,ZUO Qiang,CAI Hong-bing,et al.Clinical value of serum tissue polypeptide specific antigen detection for primary hepatic cancer[J].,2005,(05):89.
[3]郑瑞丹,杨铮,饶日春,等.酷似原发性肝癌的肝巨大血管瘤1例报告[J].南方医科大学学报,2004,(03):342.
[4]薛净,李瑜元,聂玉强,等.高强度聚焦超声治疗原发性肝癌104例临床分析[J].南方医科大学学报,2006,(11):1675.
[5]董淑英,童旭辉,蒋国君,等.星形胶质细胞中缝隙连接蛋白connexin 43的表达及其功能调控[J].南方医科大学学报,2012,(10):1423.
[6]贺松其,程旸,朱云,等.鳖甲煎丸对肝细胞癌中Wnt/β-catenin信号通路及抑制基因DKK-1、FrpHe表达的影响[J].南方医科大学学报,2013,(01):30.
[7]郭伟伟,刘莉,吴德华.DC-CIK细胞免疫治疗联合TACE术治疗原发性肝癌[J].南方医科大学学报,2014,(05):674.

更新日期/Last Update: 1900-01-01