A novel L-shaped ortho-quinone analog targeting adenosine A2b receptor to inhibit epithelial-mesenchymal transition in colorectal cancer cells.

Colorectal cancer (CRC) is a common malignancy of the gastrointestinal tract, with its incidence and mortality rates rising significantly in recent decades. In this study, we identified a compound (TC4) from a series of L-shaped ortho-quinone analog with notable inhibitory effects on epithelial-mesenchymal transition (EMT) in CRC cells. In vitro studies demonstrated that TC4 induces apoptosis, thereby suppressing CRC cell growth, invasion, and metastasis. Target analysis suggested that adenosine A2b receptor (ADORA2B) is a key molecular target of TC4, which was further confirmed by thermodynamic experiments showing direct binding to ADORA2B in living cells. Using ADORA2B overexpression and knockdown models, we found that abnormal expression of ADORA2B significantly affects CRC cell growth, invasion, metastasis, and sensitivity to TC4, confirming ADORA2B as a critical target for the compound's anti-tumor activity. TC4 was shown to markedly influence EMT, downregulating E-cadherin while upregulating N-cadherin, Vimentin, and Snail, with these effects dependent on ADORA2B overexpression. This indicates that the regulation of EMT by TC4 is closely associated with its interaction with ADORA2B. The present study confirms that TC4, a newly discovered compound with the ability to inhibit the growth and metastasis of CRC cells, can target ADORA2B to significantly regulate EMT in cancer cells.