Curcumin alters distinct molecular pathways in breast cancer subtypes revealed by integrated miRNA/mRNA expression analysis

Abstract

Background Curcumin is well known for its anticancer properties. Its cytotoxic activity has been documented in several cancer cell lines, including breast cancer. The pleiotropic activity of curcumin as an antioxidant, an antiangiogenic, antiproliferative, and pro‐apoptotic, is due to its diverse targets, such as signaling pathways, protein/enzyme, or noncoding gene. Aim This study aimed to identify key miRNAs and mRNAs induced by curcumin in breast cancer cells MCF7, T47D (hormone positive), versus MDA‐MB231 (hormone negative) using comparative analysis of global gene expression profiles. Methods RNA was isolated and subjected to mRNA and miRNA library sequencing to study the global gene expression profile of curcumin‐treated breast cancer cells. The differential expression of gene and miRNA was performed using the DESeq R package. The enriched pathways were studied using cluster profileR, and integrated miRNA–mRNA analysis was carried out using miRtarvis and miRmapper tools. Results Curcumin treatment led to upregulation of 59% TSGs in MCF7, 21% in MDA‐MB‐231 cells, and 36% TSGs in T47D, and downregulation of 57% oncogenes in MCF7, 76% in MDA‐MB‐231, and 91% in T47D. Similarly, curcumin treatment led to upregulation of 32% TSmiRs in MCF7, 37.5% in MDA‐MB231, and 62.5% in T47D, and downregulation of 77% oncomiRs in MCF7, 50% in MDA‐MB231 and 28.6% in T47D. Integrated analysis of miRNA–mRNA led to the identification of a common NFKB pathway altered by curcumin in all three cell lines. Analysis of uniquely enriched pathway revealed non‐integrin membrane–ECM interactions and laminin interactions in MCF7; extracellular matrix organization and degradation in MDA‐MB‐231 and cell cycle arrest and G2/M transition in T47D. Conclusion Curcumin regulates miRNA and mRNA in a cell type‐specific manner. The integrative analysis led to the detection of miRNAs and mRNAs pairs, which can be used as biomarkers associated with carcinogenesis, diagnostic, and treatment response in breast cancer.

ST08 Altered NF-κB Pathway in Breast Cancer Cells In Vitro as Revealed by miRNA-mRNA Analysis and Enhanced the Effect of Cisplatin on Tumour Reduction in EAC Mouse Model

“…Phorbol ester (phorbol 12- myristate 13-acetate; PMA) and IKK-16 were purchased from Tocris Bioscience (Bristol, United Kingdom) and dissolved in DMSO. The structure of ST08 and molecular signatures of each cell line are tabulated in Table 1 ( 34 ).…”

“…MDA-MB-231, MCF7 cells were incubated with different concentrations of ST08 (0,20,40,75,80,100 nM) for 48h, and western blotting was performed as described ( 2 , 5 , 34 ). The tumor tissues(100mg) from two groups (Control and ST08 treated) were minced using liquid nitrogen.…”

“…Drug Treatment, RNA isolation, and Library preparation: The experiment was performed as described ( 34 ). Briefly, 0.75 x 10^5 MDA-MB-231 cells were seeded in each well of a 6-well plate and were treated with 75nM ST08.…”

“…mRNA library preparation: As described ( 34 ), mRNAs were separated using oligo-dT beads and fragmented to 200-250 bp from the total RNA. After synthesizing cDNA, the ends were repaired for blunt ends, and the 3’ ends were adenylated.…”

“…miRNA library preparation: RNA isolation was done as mentioned above ( 34 ), and RNA sample was given for library preparation. miRNA-library preparation was outsourced to SciGenom Labs, India.…”