Midkine derived from cancer-associated fibroblasts promotes cisplatin-resistance via up-regulation of the expression of lncRNA ANRIL in tumour cells

Dongya Zhang,Yayi Hou,Jing Ren,Yanhong Ni,Shuli Zhao,Yi Li,Guoping Shi,Liang Ding,Yong Wang

doi:10.1038/s41598-017-13431-y

Dongya Zhang, Yayi Hou + Show 7 more

Open Access

https://doi.org/10.1038/s41598-017-13431-y

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Journal: Scientific Reports	Publication Date: Nov 24, 2017
Citations: 69	License type: open-access

Affiliation: Nanjing University, Nanjing Medical University

Abstract

Midkine (MK) is a heparin-binding growth factor that promotes carcinogenesis and chemoresistance. The tumour microenvironment (TME) can affect chemotherapy sensitivity. However, the role of stromal-derived MK, especially in cancer-associated fibroblasts (CAFs), is unclear. Here, we confirmed that MK decreased cisplatin-induced cell death in oral squamous cell carcinoma (OSCC) cells, ovarian cancer cells and lung cancer cells. We also isolated primary CAFs (n = 3) from OSCC patients and found that CAFs secreted increased levels of MK, which abrogated cisplatin-induced cell death. Moreover, MK increased the expression of lncRNA ANRIL in the tumour cells. Normal tissues, matched tumour-adjacent tissues and OSCC tissues were analysed (n = 60) and showed that lncRNA ANRIL was indeed overexpressed during carcinogenesis and correlated with both high TNM stage and lymph node metastasis (LNM). Furthermore, lncRNA ANRIL knockdown in tumour cells inhibited proliferation, induced apoptosis and increased cisplatin cytotoxicity of the tumour cells via impairment of the drug transporters MRP1 and ABCC2, which could be restored by treatment with human MK in a caspase-3/BCL-2-dependent manner. In conclusion, we firstly describe that CAFs in the TME contribute to the high level of MK in tumours and that CAF-derived MK can promote cisplatin resistance via the elevated expression of lncRNA ANRIL.

Highlights

As an alkylating agent, cisplatin, is one of the most effective and commonly used chemotherapeutic agents for oral squamous cell carcinoma (OSCC) and other solid tumours, including testicular, ovarian, cervical and non-small-cell lung cancer[1]
To determine whether MK-enhanced tumour cells are resistant to cisplatin, we first determined the IC50 values of several tumour cells, including the human oral squamous cell lines HSC3, OSCC3 and SCC4; the human ovarian cancer cell line A2780; and the lung cancer cell line A549 based on their differing sensitivities to cisplatin-induced cell death
We found that the cell viability of HSC3, OSCC3 and SCC4 cells with MK treatment for 24 h was increased compared to cells without MK treatment (Fig. 1a–c); the results of experiments using A2780 and A549 cells were increased (Fig. 1d,e)

Summary

Introduction

Cisplatin (cis-diamminedichloroplatinum, DDP), is one of the most effective and commonly used chemotherapeutic agents for oral squamous cell carcinoma (OSCC) and other solid tumours, including testicular, ovarian, cervical and non-small-cell lung cancer[1]. It has been reported that MK promotes tumour progression by enhancing carcinoma cell growth and survival[5,6], cell invasiveness and migration and chemotherapy resistance[7,8,9,10,11]. Activation of autophagy-mediated cell death by the Akt/mTORC1 pathway[13] All these studies focused merely on tumour-derived MK in an autocrine manner; the role of stroma-derived MK still needed to be clarified. Another study suggested that overexpression of lncRNA ANRIL was closely associated with the poor prognosis of patients with NSCLC and enhanced cell proliferation and apoptosis by binding to PRC2 to induce epigenetic silencing of KLF2 and P21 transcription[26]. The effects of lncRNA ANRIL on chemoresistance are still not well understood

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