Evaluating the cytotoxic effect of crocin on MDA-MB-۴۶۸ cell line based on apoptosis induction, ER stress, and autophagy markers

Objective: Crocin, an important saffron ingredient, showed anticancer activity in a variety of cancer types, particularly breast cancer. However, little information is available on the mechanism of its action. Previous studies indicate apoptosis induction by crocin in some cancer cells. This study aims to investigate the effect of crocin on the MDA-MB-۴۶۸ breast cancer cell line in order to investigate its effect on caspase ۹ (Cas۹) and cleaved-Cas۹, expression and splicing of XBP۱, and accumulation of LC۳-II. Methods: We used the MTT assay to investigate the cytotoxic effect of crocin on MDA-MB-۴۶۸ breast cancer cells. Next, Cas۹ and cleaved-Cas۹ levels were evaluated by Western blot analysis. Splicing of XBP۱ mRNA and expression of the spliced protein (XBP۱s) was investigated by RT-PCR and Western blot, respectively. The accumulation of LC۳-II was also evaluated by Western blot. The obtained results were analyzed and reported by Image J software. Results: The results showed a time and dose-dependent cytotoxic effect of crocin in MDA-MB-۴۶۸ cells. The expression of Cas۹ and its cleavage, therefore, the ratio of cleaved-Cas۹/Cas۹ significantly increased. Crocin treatment led to a noticeable increase in splicing of XBP۱ mRNA, expression of XBP۱s, accumulation of LC۳-II, and increased the LC۳-II/LC۳-I ratio in these cells. Conclusion: The data have shown induction of apoptosis in these breast cancer cell lines after crocin treatment. Because of the observed changes in UPR markers and autophagy, it seems that these pathways are possibly involved in this process and in intracellular regulations.

کلیدواژه ها:

Breast Cancer ، UPR ، XBP۱ splicing ، LC۳-II accumulation ، Cleaved-Cas۹ ، سرطان پستان ، کاسپاز ۹ شکسته شده ، تنش شبکه اندوپلاسمی ، پیرایش XBP۱ ، تجمع LC۳-II