Nanoquinacrine induced apoptosis in cervical cancer stem cells through the inhibition of hedgehog-GLI1 cascade: Role of GLI-1.

Anmada Nayak,Neha Tripathi,Shakti Ranjan Satapathy,Chanakyanath Kundu,Prasad V Bharatam,Sumit Siddharth,Dipon Das

doi:10.1038/srep20600

Anmada Nayak, Neha Tripathi + Show 5 more

Open Access

https://doi.org/10.1038/srep20600

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Abstract

To improve the pharmacokinetics and to study the anti-cervical cancer and anti-stem cells (CSCs) mechanism of Quinacrine (QC), a spherical nano particle of QC (i.e. NQC) was prepared and characterized. QC and NQC showed higher cytotoxicity in multiple cancer cells than the normal epithelial cells. NQC exhibited more toxicity in cervical cancer cells and its CSCs than QC. A dose-dependent decreased expression of Hedgehog-GLI (HH-GLI) components were noted in NQC treated HeLa cells and its CSCs. NQC increased the expressions of negatively regulated HH-GLI components (GSK3β, PTEN) and caused apoptosis in CSCs. Reduction of GLI1 at mRNA and promoter level were noted after NQC exposure. The expressions of HH-GLI components, GLI1 promoter activity and apoptosis were unaltered in NQC treated GLI1-knockdown cells. In silico, cell based and in vitro reconstitution assay revealed that NQC inhibit HH-GLI cascade by binding to the consensus sequence (5′GACCACCCA3′) of GLI1 in GLI-DNA complex through destabilizing DNA-GLI1 complex. NQC reduced the tumors size and proliferation marker Ki-67 in an in vivo xenograft mice model. Thus, NQC induced apoptosis in cancers through inhibition of HH-GLI cascade by GLI1. Detail interaction of QC-DNA-GLI complex can pave path for anticancer drug design.

Highlights

Metastasis is a complex process of sequential events characterized by tumor cell detachment, Epithelial to mesenchymal transition (EMT), intravasation, survival within blood and lymphatic vessel, mesenchymal to epithelial transition, micrometastasis and macrometastasis[21]
Multiple small molecule inhibitors against HH-GLI signaling are tested in multiple in vitro and in vivo systems and some of them are in clinical trial but nothing is available in clinic[29]
Due to high drug efflux, DNA repair and self renewable capabilities of cancer stem cells (CSCs), it often escapes from the toxic effect of drug and survives

Summary

Introduction

QC acts as a cytotoxic agent through a number of mechanisms which are not yet completely explored This demands identification of the target signalling pathway, as its inhibition leads to cellular apoptosis; which further requires better understanding of various constituents of cancer. SHH interaction with PTCH1 relieves the inhibition of SMO which in turn leads to the activation of GLI group of transcription factors. GANT61, a specific inhibitor of GLI1 reduces cell viability, spheroid formation, GLI-DNA binding, transcriptional activities and induces apoptosis by caspase 3 activation, PARP cleavage. It up-regulated the death receptors TRAIL-R1/DR4, TRAIL-R2/DR5 and Fas in pancreatic cancer stem cells[30,31].

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