Additive effects of Annexin A۵ overexpression in combination with ۵-Fluorouracil in reducing cell viability of Ca Ski cell

: Cervical cancer is the fourth leading cause of cancer deaths in women globally. Antimetabolite drugs work by inhibiting essential biosynthetic processes, or by being incorporated into macromolecules, such as DNA and RNA, and inhibiting their normal function. The fluoropyrimidine ۵-fluorouracil (۵-FU) does both. The mechanism of cytotoxicity of ۵-FU has been ascribed to the misincorporation of fluoronucleotides into RNA and DNA and to the inhibition of the nucleotide synthetic enzyme thymidylate synthase. ANXA۵ reportedly contributes to a variety of cancers' tumor progression, invasion, metastasis, drug resistance, and tumor treatments in certain types of cancers, hepatocarcinoma, breast cancer, cervical cancer, colorectal adenocarcinoma, gastric cancer, pancreatic adenocarcinoma, bladder cancer, nasopharyngeal carcinoma, prostate cancer, squamous cell carcinoma, glioma, sarcoma and thyroid cancer. Combining gene therapy with chemotherapy may improve cervical cancer treatment outcomes. This study evaluated the effects of Annexin A۵ (ANXA۵) overexpression alongside ۵-fluorouracil (۵-FU) on the viability of cervical squamous carcinoma (SCC) cells. Method: pAdenoVator-CMV-ANXA۵-IRES-GFP-plasmid and mock plasmid were transfected into CaSki cells using calcium-phosphate. Seventy-two hours post-transfection, GFP expression was quantified by fluorescence microscopy and flow cytometry to evaluate transfection efficiency. ANXA۵ overexpression was confirmed via qPCR. Twenty-four hours post-transfection, cells received were treated with ۱ and ۲ ug/ml of ۵-FU (IC۵۰ = ۲.۷۸۳ ug/ml).Bcl-۲ and Bax gene expression were assessed via MTT and qPCR assays. Result: ANXA۵ was overexpressed ۳۱.۵-fold compared to control (p < ۰.۰۰۰۱). MTT assays showed ANXA۵ overexpression reduced CaSki cell viability (p < ۰.۰۰۱). IC۵۰ of ۵-FU was reduced combined with ANXA۵ overexpression. Combination therapy significantly decreased Bcl-۲ expression and increased Bax versus control (p < ۰.۰۰۱). Conclusion: In conclusion, our research demonstrates that the combination of ۵-fluorouracil along with ANXA۵ overexpression, has a significant impact on inducing apoptosis and reducing cell viability in CaSki cells. These findings shed light on the potential of ANXA۵ as an anticancer protein in SCC and provide insights into its mechanisms of action. Further research is warranted to elucidate the precise role of ANXA۵ in the context of combination therapy for cervical cancer.

کلیدواژه ها:

Cervical cancer ، Annexin A۵ ، Apoptosis ، Cell cycle