Versatile Pt NCs-based chemotherapeutic agents significantly induce the apoptosis of cisplatin-resistant non-small cell lung cancer

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Highlights

  • First attempt for Pt NPs- or NCs-based chemotherapeutic agents applied in the chemotherapy for NSCLC.

  • Pt NCs-based chemotherapeutic agents significantly inhibited the proliferation of NSCLC cells in comparison with cisplatin.

  • Specifically promote cell apoptosis of cisplatin-resistant NSCLC cells.

  • Overcome the cisplatin-resistance for NSCLC.

Abstract

Recently, the incidence of lung cancer is generally rising along with air pollution and smoking, and non-small cell lung cancer (NSCLC) accounts for nearly 85% among all lung cancer diagnoses. With the development of chemotherapy, the drug resistance rate of common platinum-based chemotherapeutic drugs (like cisplatin) is gradually increased, which seriously affects the chemotherapy efficiency and survival rate of patients. In this study, polyethylenimine caged platinum nanoclusters (PEI-caged Pt NCs) were proposed as a new chemotherapeutic agent to apply in the treatment of NSCLC, choosing the classical cisplatin-resistant A549/DDP cells and normal A549 cells as targets. It was found that our Pt NCs-based chemotherapeutic drugs showed its preferable therapeutic effect in cisplatin-resistant NSCLC through the results of confocal microscopic images, cell counting kit-8 test, cell apoptosis assay and western blot. Most importantly, in the cisplatin-resistance A549/DDP cells, this kind of agents could enter the nucleus obviously, and emerged a superior inhibitory and apoptotic effects than A549 via activating p53 protein and the related signaling pathways. Comparing with the traditional chemotherapy drugs, these Pt NCs-based chemotherapeutic agents exhibit great potential and advantages in the treatment and diagnosis of NSCLC regardless of the therapeutic effect or toxic side effects, especially the drug resistance.

Graphical abstract

Pt NCs-based chemotherapeutic agent can significantly inhibit proliferation of NSCLC cells compared to cisplatin and specifically promote cell apoptosis for both A549 and drug-resistant A549/DDP cells, exhibiting the great potential and advantages in the diagnosis and treatment of NSCLC.

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Introduction

In the past decades, the incidence of lung cancer emerges an increasing trend along with air pollution and smoking which seriously affects the human survival. Lung cancer, as one of malignant cancers with high incidence, can divided into two main types: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) [1,2]. In general, NSCLC accounts for nearly 85% among all lung cancer diagnoses [3]. Surgical treatment of lung cancer is the main and effective method for the non-metastatic and early stage lung cancer, but more than two-thirds of lung cancer patients have been already diagnosed as the advanced or metastatic cancer when it was found. Chemotherapy is one of the important adjuvant therapies for lung cancer patients, which can achieve an effective palliation. Accompanying with the increase of chemotherapy drug resistance rate, the treatment of traditional chemotherapy was confronted with the technological barrier. Therefore, more potential therapeutic strategies were explored based on the cancer molecular biology, including targeting epidermal growth factor receptor (EGFR) and other signal transduction and angiogenesis pathways [4,5]. As the development of molecular profiling technology in these years, targeted therapies bring hope to various refractory lung cancer patients, which has become an innovative breakthrough in the traditional treatment of lung cancer. Regretfully, targeted therapies have not demonstrated a survival advantage in stage IV diseases via many randomized trials, even though this kind of treatments have improved survival and quality of patients distinctly [6]. Compared with targeted therapy, chemotherapy has a wider therapeutic range and better price for lung cancer patients.

Platinum (Pt) stepped into the anti-cancer drugs when it was authorized by Food and Drug Administration (FDA) in 1978, and was revealed the remarkable curative effects in cancer therapy [[7], [8], [9], [10], [11], [12]]. Nonetheless, cisplatin (in Pt +2 state), as a representative drug, possesses a number of disadvantages which weakens their therapeutic efficiency through resulting in the ototoxicity, nephrotoxicity, neurotoxicity and myelosuppression [[13], [14], [15]]. Besides, various cancers, like colorectal, prostate and breast cancer, are essential insensitive to cisplatin, while others (e.g. testicular and ovarian cancer) may emerge the resistance to cisplatin after several cycles of therapy even though these cancer cells show sensitivity, stability and efficacy at the beginning stage [[16], [17], [18]]. As for the lung cancer, cisplatin-resistance is one of the most common and intractable problems during the clinical treatment of NSCLC on account of multiple mechanism such as lack of effective drug concentration in tumor cells, reduction of drug activity, DNA repair pathways, cell apoptosis changes, and so on [19]. Some genomics studies indicated that the mutation, translocation, missing, amplification and epigenetic change of protein-coding genes or promoter area, microRNA expression changes could lead to the drug-resistance of tumor [20]. Moreover, the phenotype of tumor drug-resistance is related to the change of multiple signal pathways. At present, taxanes, gemcitabine, vinorelbine and irinotecan, have been utilized in the first-line treatment of NSCLC as the alternative for cisplatin, however, the overall effect of chemotherapy cannot be well realized, or it is difficult to achieve the desired effect in the second treatment even if the initial treatment effect is satisfied. According to statistics, the combined chemotherapy efficiency of NSCLC is only 14–40%, and it is extremely easy to relapse. Thus, the new safe and effective Pt-based chemotherapeutic agents or novel way of delivering Pt-based agents are still need to explore consistently in order to overcome the serious problems like increasing incidence, complex disease, drug resistance, etc. [[21], [22], [23]] Recently, Pt nanoparticles (NPs) or nanoclusters (NCs), were applied in the Pt-based anticancer drug system and exhibited their potential to induce the apoptosis of several cancer cells [[24], [25], [26]]. Their apoptosis mechanism is owing to the rapid decomposition by intracellular acidic organelles and then the oxidation states of Pt trends to combine with DNA or proteins resulting in the DNA destroy. Xia et al. presented Pt nanocluster assembly could damage DNA through targeting disseminated hepatocellular carcinoma (HCC) tumor-initiating cancer stem-like cells (CSLCs) to achieve the effect of inhibiting tumor cell proliferation [27]. In our previous study, dual-functional Pt NCs-based anticancer materials were employed to biological image the blood system cells as fluorescent markers and selectively inhibit K562 and BV173 cancer cells through inducing the expression of pro-apoptotic proteins (p53, PUMA, and cleaved caspase) [15]. Until to now, it was not found Pt NPs- or NCs-based chemotherapeutic agents applied in the chemotherapy for NSCLC system.

Herein, we proposed polyethyleneimine-caged Pt NCs (PEI-caged Pt NCs) to use in the treatment of NSCLC, choosing the classical cisplatin-resistant A549/DDP cells and normal A549 cells as targets. These Pt NCs-based drugs could easily enter both A549 and A549/DDP cells achieving the biological imaging of NSCLC cells as fluorescent markers. Comparing with cisplatin, our Pt NCs-based chemotherapeutic agents significantly inhibited the proliferation of NSCLC cells, in addition, they also obviously induced the apoptosis in both A549/DDP and A549 cells. Most importantly, our agents showed the obvious entering behaviors for cell nucleus, superior inhibition and apoptosis effect on the cisplatin-resistance cells. Without adding any other fluorescent biomarkers, Pt NCs-based chemotherapeutic agents can overcome the problem of drug-resistance for NSCLC system, which exhibits the promising potential in the diagnosis and treatment of NSCLC.

Section snippets

Cell culture

Human lung adenocarcinoma (LAC) cells A549 were purchased from the Cell Bank of Chinese Academy of Sciences (Shanghai, China). A549 resistance to DDP (A549/DDP) was brought from the SiXin Biotechnology Corporation Ltd. (Shanghai, China). A549/DDP cell resistance index (RI) was 4.51. After three times of cell reproduction, 2 μg/mL DDP should be added into the medium to maintain its resistance. All cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, GIBCO, New York, NY, USA) containing

Bio-imaging of NSCLC cells

The classical cisplatin-resistant A549/DDP cells and normal A549 cells were selected as targets to check the bio-imaging ability for Pt NCs-based drugs (Fig. 1a). The confocal microscopic images of A549 and A549/DDP cells labeled with Pt NCs-based chemotherapeutic agent are presented in Fig. 2. Both A549 and A549/DDP cells exhibit the red fluorescence signal that is emitted from our Pt NCs-based chemotherapeutic agent, while the cell nuclei are stained by DAPI showing the blue fluorescence.

Discussion

Based on the previous studies, specific DNA damages that induce apoptosis have been identified, which is one of the important routes of cell inactivation [22]. It is well known that P53 is a vital portion in the apoptotic response of cells, which induces transcriptional activation of pro-apoptotic factors such as recombinant factor related apoptosis (FAS), recombinant p53 upregulated modulator of apoptosis (PUMA) and Bcl-2 associated X protein (BAX) [28,29]. What has been confirmed is that a

Acknowledgements

This research was funded by National Natural Science Foundation of China (21807121), Key Scientific Research Projects of High Education of Henan Province (18A430005), Medical Science and Technology Project of Henan Province (201503133), and Project for Fundamental Research Funds of Zhongyuan University of Technology (K2018YY020). Dr. X. Huang gratefully acknowledges the support from “The 2019 Youth Talents Promotion Project of Henan Province”, “The 2018 Backbone Teachers of Zhongyuan University

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    These authors contributed equally to this work and should be considered as first authors.

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