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Original Research

Ischemia Induces Quiescence and Autophagy Dependence in Hepatocellular Carcinoma

Published Online:Mar 2 2017https://doi.org/10.1148/radiol.2017160728

Abstract

Our findings, in combination with the nutrient deprivation and oxidative stress associated with the post-transarterial embolization (TAE)/transarterial chemoembolization (TACE) microenvironment, suggest that autophagy may represent an optimal and even necessary target for adjunct therapy in TAE/TACE.

Purpose

To characterize hepatocellular carcinoma (HCC) cells surviving ischemia with respect to cell cycle kinetics, chemosensitivity, and molecular dependencies that may be exploited to potentiate treatment with transarterial embolization (TAE).

Materials and Methods

Animal studies were performed according to institutionally approved protocols. The growth kinetics of HCC cells were studied in standard and ischemic conditions. Viability and cell cycle kinetics were measured by using flow cytometry. Cytotoxicity profiling was performed by using a colorimetric cell proliferation assay. Analyses of the Cancer Genome Atlas HCC RNA-sequencing data were performed by using Ingenuity Pathway Analysis software. Activation of molecular mediators of autophagy was measured with Western blot analysis and fluorescence microscopy. In vivo TAE was performed in a rat model of HCC with (n = 5) and without (n = 5) the autophagy inhibitor Lys05. Statistical analyses were performed by using GraphPad software.

Results

HCC cells survived ischemia with an up to 43% increase in the fraction of quiescent cells as compared with cells grown in standard conditions (P < .004). Neither doxorubicin nor mitomycin C potentiated the cytotoxic effects of ischemia. Gene-set analysis revealed an increase in mRNA expression of the mediators of autophagy (eg, CDKN2A, PPP2R2C, and TRAF2) in HCC as compared with normal liver. Cells surviving ischemia were autophagy dependent. Combination therapy coupling autophagy inhibition and TAE in a rat model of HCC resulted in a 21% increase in tumor necrosis compared with TAE alone (P = .044).

Conclusion

Ischemia induces quiescence in surviving HCC cells, resulting in a dependence on autophagy, providing a potential therapeutic target for combination therapy with TAE.

© RSNA, 2017

Online supplemental material is available for this article.

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Article History

Received April 5, 2016; revision requested June 15; revision received October 12; accepted November 21; final version accepted December 18.
Published online: Mar 02 2017
Published in print: June 2017