Dihydroartemisinin Sensitizes Mutant p53 (R248Q)-Expressing Hepatocellular Carcinoma Cells to Doxorubicin by Inhibiting P-gp Expression.

Yue Yang,Yun Su,Jianxin He,Jing Chen,Hulai Wei,Li Lin,Cunmin Zhou,Yongqi Liu

doi:10.1155/2019/8207056

Abstract

Mutant p53 (R248Q) induces doxorubicin (ADM) resistance in hepatocellular carcinoma (HCC). Dihydroartemisinin (DHA) can synergistically enhance anticancer effect of many chemotherapeutic agents. However, whether DHA could increase therapeutic efficacy of ADM in p53 (R248Q)-expressing HCC cells remains unknown. In the present study, we established mutant p53 (R248Q)-expressing Hep3B cells to study the effect and mechanism of DHA on ADM resistance and the synergistic effect of DHA with ADM. We found that P-gp was highly expressed in p53 (R248Q)-expressing Hep3B cells. As a result, cells expressing p53 (R248Q) displayed higher cell viability and lower cell apoptosis level upon ADM treatment. Meanwhile, phosphorylation levels of ERK1/2 and p65 were elevated in p53 (R248Q)-expressing Hep3B cells. However, combination of DHA and ADM treatment decreased cell viability and elevated cell apoptosis level in p53 (R248Q)-expressing Hep3B cells. Molecular dynamics simulations showed that DHA had the potential to bind with mutant p53 (R248Q) protein. Furthermore, DHA treatment decreased P-gp expression and inhibited phosphorylation levels of ERK1/2 and p65 in p53 (R248Q)-expressing Hep3B cells. Finally, DHA treatment could significantly reduce ADM efflux in p53 (R248Q)-expressing cells. Our results indicate that DHA could decrease P-gp expression via inhibiting the p53 (R248Q)-ERK1/2-NF-κB signaling pathway, which eventually confers sensitization of p53 (R248Q)-expressing HCC cells to ADM. Our study provides evidence for the potential application of DHA and ADM combination in treatment of mutant p53 (R248Q)-harbored HCC.

Highlights

Liver cancer is currently the third leading cause of cancerrelated mortality worldwide [1, 2]
We further demonstrated that DHA decreased the expression of P-gp via inhibiting p53 (R248Q)ERK1/2-NF-κB signaling, which leads to the reversal of ADM resistance in p53 (R248Q)-expressing Hepatocellular carcinoma (HCC) cells
In order to analyze the effects of p53 (R248Q) on P-gp expression, we firstly constructed Hep3B cells expressing mutant p53 (R248Q) and detected P-gp expression in Hep3B-derived cells. p53 (R248Q)expressing cells showed obviously increased P-gp expression compared with either empty vector lenti-virus infected cells or WT p53-expressing cells (Figures 1(a), 1(b)). en, we examined the ADM resistance in p53expressing cells by detecting cell survival after ADM treatment. e results showed that cells expressing p53 (R248Q) exhibited significantly higher survival in comparison with either control cells or WT p53-expressing cells upon ADM treatment (Figure 1(c))

Summary

Introduction

Liver cancer is currently the third leading cause of cancerrelated mortality worldwide [1, 2]. Hepatocellular carcinoma (HCC) is the most common type of liver cancer [3]. Chemotherapy is a major treatment for HCC patients because most patients are diagnosed at advanced stages when the surgical excision is infeasible [4]. Doxorubicin (ADM) is a commonly used drug for HCC [5]. Long-term use of ADM inevitably causes drug resistance and greatly reduces the treatment efficacy in HCC patients [6]. P-gp belongs to the family of ATPdependent membrane pumps, which can facilitate the efflux of chemotherapeutic drugs such as ADM, vincristine, and paclitaxel, resulting in drug resistance [9,10,11]

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Results

Conclusion