Investigating the effect of Auraptene in colorectal cancer cell lines (SW۴۸۰ and HT۲۹)

introduction: Colon or colorectal cancer (CRC) is the third most common cancer in the world. The most deadly cancer is colon cancer because it is resistant to drugs and leads to metastasis and recurrence. Colon cancer is one of the most dangerous malignancies. Cancer cells' method for survival is to escape from apoptosis and uncontrolled cell division. One of the approaches to cancer treatment is the use of anti-cancer agents that induce apoptosis and control cell division. Despite new methods in adjuvant chemotherapy, cancer cell metastasis is a global concern. In adjuvant treatment for CRC, herbal medicines with minimal side effects may be better choices. For treatment of malignancies, herbal medicines with anti-tumor effects are a suitable alternative. Coumarins with anti-cancer activity against different types of cancer can act as a powerful adjuvant therapy in CRC because it causes apoptosis and cell cycle arrest. Because Auraptene is an abundant natural coumarin, we decided to investigate the therapeutic effects of Auraptene on SW۴۸۰ and HT۲۹ colorectal cancer (CRC) cells. We tested the antiproliferative and cytotoxicity effects of Auraptene by ۳-۴,۵-dimethylthiazol-۲-yl-۲,۵ diphenyltetrazolium bromide (MTT) test, and to determined cell migration used scratch test and to check apoptosis and necrosis was used Annexin V/ PI method. Finally, Real-Time PCR was used to determine the expression levels of mir۲۷a۳p after Auraptene treatment. Materials and Methods: The IC۵۰ values of Auraptene were calculated in SW۴۸۰ and HT۲۹ Colorectal cancer cells by treating them with serial dilutions of Auraptene for ۲۴,۴۸ and ۷۲ hours, respectively. The cell viability was assessed through the MTT assay. Subsequently, cells were divided into two groups: Auraptene (cells treated with auraptene) and control (cells not treated). The wound healing assay (or scratch) to monitor cell proliferation and cell migration. Using flow cytometry, the annexin V protein was shown to initiate the apoptosis and necrosis experiment. Also, The study utilized real-time PCR to evaluate the expression of the MIR۲۷a-۳p. Results: The study found that Auraptene significantly inhibited the survival of SW۴۸۰ and HT۲۹ cells, with IC۵۰ values of ۲۰۰ μg/μL at ۲۴, ۴۸ and ۷۲ hours for SW۴۸۰ and ۱۵۰ μg/μL at ۴۸ and ۷۲ hours for HT۲۹, respectively. The flow cytometry study confirmed that Auraptene induced apoptosis and necrosis in SW۴۸۰ and HT۲۹ cells. Moreover, Real-Time PCR revealed significantly higher expression of MIR۲۷a-۳p in the control group compared to the auraptene group. The results of the investigation showed that Auraptene dramatically reduced the ability of SW۴۸۸۰ and HT۲۹ cells to survive; the IC۵۰ values for SW۴۸۰ were ۲۰۰ μg/μL at ۲۴, ۴۸, and ۷۲ hours, and for HT۲۹ they were ۱۵۰ μg/μL at ۴۸ and ۷۲ hours. The investigation conducted using flow cytometry techniques verified that auraptene caused apoptosis and necrosis in both HT۲۹ and SW۴۸۰ cells. Conclusion: This study showed that Auraptene causes apoptosis and could be used as an anticancer medication for colon cancer treatment with further research.

کلیدواژه ها:

Apoptosis ، Auraptene ، Colorectal cancer ، SW۴۸۰ cell line ، HT۲۹ cell line