Piperine sensitizes radiation‐resistant cancer cells towards radiation and promotes intrinsic pathway of apoptosis

Piperine, a bioactive alkaloid, is known to have anticancer activities. Hence, in this study, the effectiveness of piperine pretreatment as a strategy for radio‐sensitizing colorectal adenocarcinoma cell line (HT‐29) was analyzed. For this, HT‐29 cells were pretreated with piperine (12.5 and 25 µg/mL) and exposed to γ‐radiation (1.25 Gy) and analyzed for various effector pathways to elucidate the possible mode of action in comparison to individual treatments. The proliferation efficiency of the cells was analyzed by trypan blue dye exclusion assay and MTT assay. The synergistic effects of the combination treatment were analyzed with compuSyn software. Downstream signaling pathways leading to apoptosis were studied using flowcytometry, immunofluorescence, and immunoblot assays. It was observed that combination treatment arrested HT‐29 cells at G2/M phase nearly 2.8 folds higher than radiation treatment alone, inducing the radio‐resistant cells to undergo apoptosis through mitochondria‐dependent pathway. In addition, activation of caspase‐3 and cleavage of poly(ADP‐ribose) polymerases‐1, the key molecular events in apoptotic signaling, were significantly enhanced. Activation of estrogen receptor beta (ERβ), a nuclear hormone transcription factor promoting tumor suppression represents a novel clinical advance towards management and prevention of cancers. Interestingly, the expression of ERβ was increased in the cells treated with piperine. In conclusion, piperine pretreatment enhances radio‐sensitization in HT‐29 cells by inducing the cells to undergo apoptosis hence, can be used as a classic candidate for colon cancer sensitization towards radiotherapy. Practical Application Piperine induces enhanced radiosensitization of colon cancer cell line (HT‐29) by interfering with the cancer cell line proliferation, DNA damage, and apoptosis.