The effect of compound kushen injection on cancer cells: Integrated identification of candidate molecular mechanisms

Abstract

Traditional Chinese Medicine (TCM) preparations are often extracts of single or multiple herbs containing hundreds of compounds, and hence it has been difficult to study their mechanisms of action. Compound Kushen Injection (CKI) is a complex mixture of compounds extracted from two medicinal plants and has been used in Chinese hospitals to treat cancer for over twenty years. To demonstrate that a systematic analysis of molecular changes resulting from complex mixtures of bioactives from TCM can identify a core set of differentially expressed (DE) genes and a reproducible set of candidate pathways. We used in vitro cancer models to measure the effect of CKI on cell cycle phases and apoptosis, and correlated those phenotypes with CKI induced changes in gene expression. We treated two cancer cell lines with or without CKI and assessed the resulting phenotypes by employing cell viability and proliferation assays. Based on these results, we carried out high-throughput transcriptome data analysis to identify genes and candidate pathways perturbed by CKI. We integrated these differential gene expression results with previously reported results and carried out validation of selected differentially expressed genes. CKI induced cell-cycle arrest and apoptosis in the cancer cell lines tested. In these cells CKI also altered the expression of 363 core candidate genes associated with cell cycle, apoptosis, DNA replication/repair, and various cancer pathways. Of these, 7 are clinically relevant to cancer diagnosis or therapy, 14 are cell cycle regulators, and most of these 21 candidates are downregulated by CKI. Comparison of our core candidate genes to a database of plant medicinal compounds and their effects on gene expression identified one-to-one, one-tomany and many-to-many regulatory relationships between compounds in CKI and DE genes. By identifying genes and promising candidate pathways associated with CKI treatment based on our transcriptome-based analysis, we have shown that this approach is useful for the systematic analysis of molecular changes resulting from complex mixtures of bioactives.

High Throughput Transcriptome Data Analysis and Computational Verification Reveal Immunotherapy Biomarkers of Compound Kushen Injection for Treating Triple-Negative Breast Cancer

“…Multiple studies have revealed the mechanism of CKI on TNBC. CKI strongly reduces the migration and invasion of MDA-MB-231 cells and induces cell cycle arrest and apoptosis in them (36,37,63,64). Furthermore, CKI has been found to suppress energy metabolism and DNA repair pathways in MDA-MB-231 cells (63).…”

Analysis of Genetic Diversity in the Traditional Chinese Medicine Plant ‘Kushen’ (Sophora flavescens Ait.)

“…Of particular importance is the use of kushen extracts in combination with extracts from Smilax glabra in compound kushen injection (CKI). CKI has been used in Chinese hospitals to treat cancer for over 20 years ( Cui et al, 2020 ) and has been reported to boost immunity, decrease inflammation, and decrease metastasis ( Wang et al, 2015 ).…”

A System Pharmacology Model for Decoding the Synergistic Mechanisms of Compound Kushen Injection in Treating Breast Cancer

“…CKI is a TCM formulae extract from kushen ( Radix Sophorae flavescentis ) and baituling ( Rhizoma Heterosmilacis ) at a ratio of 7:3, in which exist hundreds of components including alkaloids and flavonoids ( Cui et al, 2020 ). As approved by the China Food and Drug Administration, CKI can be employed for cancer treatment ( Guo et al, 2015 ) and has several pharmacological functions, including anticancer properties, hemostasis, and immunity enhancement ( Wang et al, 2015 ).…”