Novel regulation of miR‐34a‐5p and HOTAIR by the combination of berberine and gefitinib  leading to inhibition of EMT in human lung cancer

Abstract

HOTAIR is an important carcinogenic lncRNA and involves in tumorigenesis, and invasion. MiR‐34a‐5p functions as a tumour suppressor. However, the underlying mechanism of HOTAIR regulation especially in association with miR‐34a‐5p in non‐small‐cell lung cancer (NSCLC) has not been explored. Herein, we performed series of in vitro experiments, including viability, migration, invasion, apoptosis and in vivo xenograft model, and identified that HOTAIR was remarkably elevated in NSCLC cells. Enforced HOTAIR expression promoted migration and invasion, while depleted HOTAIR diminished the ability of migration and invasion of NSCLC cells. We also observed that miR‐34a‐5p was dramatically inhibited in NSCLC cells and the binding correlation between HOTAIR and miR‐34a‐5p was confirmed by dual‐luciferase reporter and RNA immunoprecipitation assays. We also showed that induction of miR‐34a‐5p and reduction of HOTAIR, and the interaction between miR‐34a‐5p and HOTAIR resulted in the suppression of epithelial‐mesenchymal transition (EMT) as illustrated by induction of key epithelial markers E‐cadherin expression, reduction of vimentin and EMT‐inducing transcription factor snail. Excessive expression of snail resisted miR‐34a‐5p‐inhibited cell growth. Snail binds to E‐cadherin promoter and regulates E‐cadherin expression. There was a synergy in combination of berberine and gefinitib in this process. Similar findings were also observed in a tumour xenograft model. Collectively, this is the first report demonstrating reciprocal interaction of miR‐34a‐5p‐ and HOTAIR‐mediated regulation of snail resulting in inhibition of EMT process by the combination of berberine and gefitinib suggesting that regulation of miR‐34a‐5p‐ and HOTAIR‐mediated inhibition of EMT may provide novel treatment paradigms for lung cancer.

Regulation of Long Non-Coding RNAs by Plant Secondary Metabolites: A Novel Anticancer Therapeutic Approach

“…Consequently, berberine indirectly reduces vimentin, and increases E-cadherin via miR-34a-5p upregulation. Interestingly, the ectopic expression of HOTAIR will eliminate the effect of berberine on EMT [ 27 ]. Considering berberine can reduce HOTAIR expression in vitro and in vivo, it has the potential to be an appropriate treatment for cancers such as NSCLC, where this lncRNA is increased.…”

“…For example, curcumin by upregulation of miR-185 and miR-29a can decrease the expression of DNMT1, DNMT 3A, and DNMT 3B to upregulate MEG3 in hepatocellular cancer cells [ 132 ]. Moreover, curcumin via upregulation of miR-145 or berberine and genistein via upregulation of miR-34a-5p could decrease the lncRNA-ROR and HOTAIR, respectively [ 27 , 127 , 143 ]. Genistein can upregulate miR-141 expression, and miR-141 has a binding site in HOTAIR.…”

“…These compounds could regulate the expression of lncRNA directly and indirectly without any side effects [ 23 , 24 ]. In vivo and in vitro studies reveal that phytochemicals could inhibit proliferation, invasion, migration, EMT, metastasis and induce chemosensitization and radiosensitization of cancer cells by downregulation of oncogenic lncRNAs or upregulation of tumor suppressor lncRNAs [ 25 , 26 , 27 ].…”

Biological Activity of Berberine—A Summary Update

“…The apoptosis was caspase-dependent, while the necroptosis accompanied the activation of the RIPK3-MLKL pathway [ 100 ]. In combination with gefitinib in non-small-cell lung cancer, berberine inhibits epithelial–mesenchymal transition [ 101 ].…”

Anti-Tumor Mechanisms Associated With Regulation of Non-Coding RNA by Active Ingredients of Chinese Medicine: A Review

“…Previous studies have found that the abnormal expression of miRNAs can disrupt many signaling pathways resulting in reduced proliferation, migration, and invasion of cancer cells and the promotion of apoptosis ( 19 – 21 ). Numerous carcinogenic miRNAs, including miR-638, miR-155, miR-31, miR-21, miR-221, miR-222, and miR-294 among others, are overexpressed in various malignant tumors ( 22 – 24 ), whereas others, including miR-429, miR-211, miR-1271, and miR-34a, exert anti-cancer effects and are under-expressed in cancer cells ( 25 , 26 ).…”