Transcriptome profiling reveals Silibinin dose-dependent response network in non-small lung cancer cells.

Jagan Mohan Kaipa,Vytaute Starkuviene,Holger Erfle,Evgeny Gladilin,Roland Eils

doi:10.7717/peerj.10373

Abstract

Silibinin (SIL), a natural flavonolignan from the milk thistle (Silybum marianum), is known to exhibit remarkable hepatoprotective, antineoplastic and EMT inhibiting effects in different cancer cells by targeting multiple molecular targets and pathways. However, the predominant majority of previous studies investigated effects of this phytocompound in a one particular cell line. Here, we carry out a systematic analysis of dose-dependent viability response to SIL in five non-small cell lung cancer (NSCLC) lines that gradually differ with respect to their intrinsic EMT stage. By correlating gene expression profiles of NSCLC cell lines with the pattern of their SIL IC50 response, a group of cell cycle, survival and stress responsive genes, including some prominent targets of STAT3 (BIRC5, FOXM1, BRCA1), was identified. The relevancy of these computationally selected genes to SIL viability response of NSCLC cells was confirmed by the transient knockdown test. In contrast to other EMT-inhibiting compounds, no correlation between the SIL IC50 and the intrinsic EMT stage of NSCLC cells was observed. Our experimental results show that SIL viability response of differently constituted NSCLC cells is linked to a subnetwork of tightly interconnected genes whose transcriptomic pattern can be used as a benchmark for assessment of individual SIL sensitivity instead of the conventional EMT signature. Insights gained in this study pave the way for optimization of customized adjuvant therapy of malignancies using Silibinin.

Highlights

The ability of malignant cells to attain drug resistance and to escape cell death frequently observed in different types of cancer represents the major challenge to chemical tumor therapy
As one can see from the overview of SIL/WFA IC50 ranks of all five non-small cell lung cancer (NSCLC) cell lines in Fig. 2, the pattern of WFA IC50 values exhibits a strong correlation with the intrinsic epithelial-mesenchymal transition (EMT) stage of NSCLC cells, while SIL IC50 does not show such a correlation
Even though the reverse transfection method was considered to be more efficient providing higher transfection rates with minimal nucleic acid usage (Erfle et al, 2007), here we found out forward transfection to show a better performance crossover five tested NSCLC cell lines, see Fig. S1

Summary

Introduction

The ability of malignant cells to attain drug resistance and to escape cell death frequently observed in different types of cancer represents the major challenge to chemical tumor therapy. Several evolutionary conserved mechanisms mediate elevated survival capabilities. Transcriptome profiling reveals Silibinin dosedependent response network in non-small lung cancer cells. As a consequence of elevated environmental adaptation and self-reprogramming capabilities, cancer cells often attain resistance against targeted drugs and even drug combinations by bypassing affected pathways (Patel & Rothenberg, 1994; Clarke et al, 2019). In most of the tumors, multiple survival mechanisms and pathways are active in parallel. A network of dozens tightly interconnected genes rather than just few linear signaling pathways maintain abnormal survival and resistance capabilities of cancer cells (Gladilin & Eils, 2017)

Methods

Results

Conclusion