Effect of apigenin on whole transcriptome profile of TNFα‑activated MDA‑MB‑468 triple negative breast cancer cells

Abstract

The lack of hormone receptors in triple negative breast cancer (TNBC) is associated with the inefficacy of anti-estrogen chemotherapies, leaving fewer options for patient treatment and higher mortality rates. Additionally, as with numerous types of inflammatory breast cancer, infiltration of tumor associated macrophages and other leukocyte sub-populations within the tumor inevitably lead to aggressive, chemo-resistant, metastatic and invasive types of cancer which escape immune surveillance. These processes are orchestrated by the release of potent cytokines, including TNFα, IL-6 and CCL2 from the stroma, tumor and immune cells within the tumor microenvironment. The present study evaluated apigenin modulating effects on the pro-inflammatory activating action of TNFα in TNBC MDA-MB-468 cells, derived from an African American woman. Initially, cell viability was determined to establish an optimal sub-lethal dose of TNFα and apigenin in MDA-MB-468 cells. Subsequently, various treatments effects were evaluated using whole transcriptomic analysis of mRNA and long intergenic non-coding RNA with Affymetrix HuGene-2.1-st human microarrays. Gene level differential expression analysis was conducted on 48,226 genes where TNFα caused significant upregulation of 53 transcripts and downregulation of 11 transcripts. The largest upward differential shift was for CCL2 [+61.86 fold change (FC); false discovery rate (FDR), P<0.0001]; which was down regulated by apigenin (to +10.71 FC vs. Control; FDR P-value <0.001), equivalent to an 83% reduction. Several TNFα deferentially upregulated transcripts were reduced by apigenin, including CXCL10, C3, PGLYRP4, IL22RA2, KMO, IL7R, ROS1, CFB, IKBKe, SLITRK6 (a checkpoint target) and MMP13. Confirmation of CCL2 experimentally induced transcript alterations was corroborated at the protein level by ELISA assays. The high level of CCL2 transcript in the cell line was comparable to that in our previous studies in MDA-MB-231 cells. The differential effects of TNFα were corroborated by ELISA, where the data revealed a >10-fold higher releasing rate of CCL2 in MDA-MB-468 cells compared with in MDA-MB-231 cells, both of which were attenuated by apigenin. The data obtained in the present study demonstrated a high level of CCL2 in MDA-MB-468 cells and a possible therapeutic role for apigenin in downregulating TNFα-mediated processes in these TNBC cells.

Low CFB expression is independently associated with poor overall and disease‑free survival in patients with lung adenocarcinoma

“…Recent studies have reported that CFB is associated with the prognosis of different types of cancer (32)(33)(34). A previous study has demonstrated that CFB is not only identified in all crypts in the colonic mucosa, but is also expressed in adenomas and carcinomas (16).…”

Targeting AMPK Signaling by Dietary Polyphenols in Cancer Prevention

“…[ 37 ] Apigenin could effectively reverse the cancer‐promoting effects of tumor necrosis factor‐α (TNF‐α) in breast cancer cell MDA‐MB‐231. [ 38,39 ] Apigenin also could promote the apoptosis of cisplatin‐resistant ovarian cancer cells (SKOV3 and DDPs) by targeting Mcl‐1 in a mitochondrial‐dependent way. [ 40 ] Growing evidences indicate that apigenin exerts anti‐cancer effect via different mechanisms in the various cancer models.…”

Identification of SLITRK6 as a Novel Biomarker in hepatocellular carcinoma by comprehensive bioinformatic analysis

“…In the study of apigenin regulating the pro-inflammatory activation of TNFα in MD-MB-468 cells of Triple-Negative Breast Cancer (TNBC). Several TNFα deferentially upregulated transcripts were reduced by apigenin, including CXCL10 , C3 , PGLYRP4 , IL22RA2 , KMO , IL7R , ROS1 , CFB , IKBKe , SLITRK6 (a checkpoint target), and MMP13 [ 46 ]. The SLITRK1 , SLITRK2 , SLITRK3 , SLITRK4 , SLITRK5 , and SLITRK6 genes have been detected in the human brain, and the expression profile of each SLITRK is unique.…”