丹参酮IIA通过调控自噬抑制乏氧诱导的肝内胆管细胞癌侵袭能力的研究
Tanshinone IIA Iinhibit the migration and invasion of Intrahepatic Cholangiocarcinoma cells through regulating the autophagy induced by hypoxia

成薇婷,许凯△.丹参酮IIA通过调控自噬抑制乏氧诱导的肝内胆管细胞癌侵袭能力的研究[J].内科急危重症杂志,2016,22(5):

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英文关键词:Intrahepatic cholangiocarcinoma hypoxia Tanshinone IIA autophagy invasion

基金项目:湖北省自然基金

作者	单位	E-mail
成薇婷	华中科技大学同济医学院附属中西医结合医院肿瘤科	joycvt@163.com
许凯△	华中科技大学同济医学院附属同济医院骨科

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中文摘要:

目的探讨在乏氧状态下，丹参酮IIA对肝内胆管细胞癌(Intrahepatic Cholangiocarcinoma, ICC)自噬的调控，以及对侵袭能力的影响作用及其机制。方法通过1%O2培养建立肝内胆管细胞癌ICC-9810细胞乏氧状态，应用丹参酮IIA干预细胞后，通过划痕试验检测ICC-9810细胞运动能力，迁移试验及侵袭试验检测细胞的迁移与侵袭能力；转染pGFP-LC3质粒后，应用荧光显微镜观察LC3融合蛋白及自噬小体在ICC-9810细胞中的表达情况，蛋白质印迹法检测HIF-1α、自噬相关蛋白LC3II/I和Beclin1在ICC-9810细胞中的表达差异。结果在乏氧环境下，ICC-9810细胞运动能力增强了79.6%，侵袭力增强了1.36倍，细胞自噬水平明显升高，丹参酮IIA干预后ICC-9810细胞自噬水平下调， HIF-1α、LC3II/I和Beclin1蛋白表达水平分别降低3.9倍，2.4倍和1.1倍，迁移和侵袭能力下调了4.7倍。结论乏氧可以促进肝内胆管细胞癌自噬水平并增强细胞运动侵袭能力，丹参酮IIA可以通过下调自噬，抑制肝内胆管癌细胞在乏氧条件下异常增高的的侵袭力。

英文摘要:

Objective: To study the effect and mechanism of Tanshinone IIA on autophagy and invasiveness of intrahepatic cholangiocarcinoma(ICC) under hypoxic condition. Methods: The hypoxia state was induced by hypoxia culture(1%) in ICC-9810 cells and then treated cells with Tanshinone IIA; scratch assay was used to test the mobility of ICC-9810 cells, migration assay and invasion assay were performed to detect the migration ability and invasiveness of ICC-9810 cells; the autophagosome and expression of LC3 fusion proteinwas observed by fluorescence microscopy in ICC-9810 cells after transfection pGFP-LC3 plasmid; Expression of HIF-1α,autophagy related protein LC3 II/I and Beclin1 were detected by Western blot. Result: Hypoxia can enhance the mobility and invasiveness for 79.6% and 1.36 fold compare to the control cells as same as autophagy of ICC-9810 cells. After stimulation with Tanshinone IIA, the level of autophagy of ICC-9810 cells was reduced, and the expression of HIF-1α, LC3 II/I and Beclin1 were down-regulated to 3.9, 2.4 and 1.1 fold respectively, at the same time, the ability of migration and invasive decreased. Conclusion: The level of autophagy and invasiveness of ICC cells can be promoted by hypoxia; Tanshinone IIA can reduce the aberrant invasiveness of ICC cells under hypoxic conditions through alleviating the level of autophagy.