Effect of Compound Kushen Injection, a Natural Compound Mixture, and Its Identified Chemical Components on Migration and Invasion of Colon, Brain, and Breast Cancer Cell Lines

Abstract

Traditional Chinese Medicines are promising sources of new agents for controlling cancer metastasis. Compound Kushen Injection (CKI), prepared from medicinal plants Sophora flavescens and Heterosmilax chinensis , disrupts cell cycle and induces apoptosis in breast cancer; however, effects on migration and invasion remained unknown. CKI, fractionated mixtures, and isolated components were tested in migration assays with colon (HT-29, SW-480, DLD-1), brain (U87-MG, U251-MG), and breast (MDA-MB-231) cancer cell lines. Human embryonic kidney (HEK-293) and human foreskin fibroblast (HFF) served as non-cancerous controls. Wound closure, transwell invasion, and live cell imaging showed CKI reduced motility in all eight lines. Fractionation and reconstitution of CKI demonstrated combinations of compounds were required for activity. Live cell imaging confirmed CKI strongly reduced migration of HT-29 and MDA-MB-231 cells, moderately slowed brain cancer cells, and had a small effect on HEK-293. CKI uniformly blocked invasiveness through extracellular matrix. Apoptosis was increased by CKI in breast cancer but not in non-cancerous lines. Cell viability was unaffected by CKI in all cell lines. Transcriptomic analyses of MDA-MB-231indicated down-regulation of actin cytoskeletal and focal adhesion genes with CKI treatment, consistent with observed impairment of cell migration. The pharmacological complexity of CKI is important for effective blockade of cancer migration and invasion.

Understanding the Mechanistic Contribution of Herbal Extracts in Compound Kushen Injection With Transcriptome Analysis

“…Based on our previous publications, perturbed regulation of genes in these pathways and annotated by these terms was associated with the observed cell viability and apoptotic effects CKI on cancer cells (10, 14). Other terms showing perturbation of metabolic processes and cell migration identified in this study, such as “pyrimidine metabolism,” “steroid biosynthesis,” and “positive regulation of locomotion,” also showed up in our previous research on the effects of CKI (25). Altogether, these results indicated that Kushen was very important to the major molecular consequences of CKI treatment, and perturbed most of the biological functions perturbed by CKI, resulting in reduced viability and and increased apoptosis in cancer cells.…”

A systems pharmacology approach to determine the mechanisms of action of pleiotropic natural products in breast cancer from transcriptome data

“…These pathways regulate cell proliferation and apoptosis (P53, RTK-RAS-MAPK, PI3K-Akt-mTOR and NRF2) and metastasis/invasion (TGF-beta). Moreover, CKI also targets angiogenesis and tumor microenvironment regulating pathways through VEGFA/VEGFR2 and cytokine signaling (B cell receptor, T cell receptor and FC-epsilon signaling) respectively ( Supplementary Table 5 ), which is consistent with a previous finding 52 . Other reports have shown that CKI directly regulates cell migration 53 ; and apoptosis in breast cancer 52 .…”

“…Moreover, CKI also targets angiogenesis and tumor microenvironment regulating pathways through VEGFA/VEGFR2 and cytokine signaling (B cell receptor, T cell receptor and FC-epsilon signaling) respectively ( Supplementary Table 5 ), which is consistent with a previous finding 52 . Other reports have shown that CKI directly regulates cell migration 53 ; and apoptosis in breast cancer 52 . Cell cycle, proliferation and apoptosis, metastasis/invasion, and angiogenesis were the main targeted carcinogenesis processes in this cell line ( Table 3 ).…”

Fractional Deletion of Compound Kushen Injection Indicates Cytokine Signaling Pathways are Critical for its Perturbation of the Cell Cycle

“…Previous studies have demonstrated that CKI can alter the cell-cycle, induce apoptosis and reduce proliferation and migration in various cancer cell lines 6,16–18 . CKI also killed leukaemia cells via the Prdxs/ROS/Trx1 signalling pathway in an acute myeloid leukaemia patient-derived xenograft model and caused cell-cycle arrest in U937 leukaemia-derived cells 19 .…”