In vitro and in silico evaluation of black wattle proanthocyanidins in bladder cancer T24 cells: gene expression profiling and computational target identification

Bladder cancer (BC) is aggressive and difficult to treat, with chemotherapy facing challenges like drug toxicity and resistance. Proanthocyanidins (PAC) show promising antitumor activity, as demonstrated by studies in several cancer types. However, much is still unknown about how PAC could be used as therapeutic adjuvants in BC. This study evaluated the antiproliferative potential of PAC from the black wattle ( Acacia mearnsii ) bark extract (BWP) in BC T24 cells using in vitro and in silico approaches. BWP treatment reduced T24 cell viability in a dose-dependent manner, with an IC 50 value of 11.67 µg/mL. Flow cytometry revealed BWP-induced apoptosis in T24 cells. Additionally, nuclear morphometric analysis indicated a shift toward a senescence-like phenotype, consistent with morphological features of senescent cells. Using an in silico systems biology approach, we identified candidate genes potentially associated with BWP activity, including BACE1, DNMT1, NOS1, NOS2, NOS3, PPARA, PPARG, PPARGC1A, and PTGS2. The mRNA expression analysis (qRT-PCR) showed that BWP modulated the expression of DNMT1, BACE1, PPARG, and NOS3 in T24 cells. suggesting potential therapeutic roles in cancer progression. In conclusion, BWP exhibits antiproliferative activity and induces apoptosis in T24 bladder cancer cells, and its constituent compounds interact with candidate genes relevant to cancer biology. These findings provide a hypothesis-generating framework for future mechanistic and functional studies on PAC-based therapies.