Research on the Synergistic Effect of A Novel Apoptosis Inducer Combined with AKT and HSP90 Selective Inhibitors on Hormone-Sensitive and Hormone-Resistant Breast Cancer Cell Lines

新型凋亡诱导剂联合AKT和HSP90选择性抑制剂对激素敏感型和激素抵抗型乳腺癌细胞系的协同作用研究

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Abstract

One of the frequent malignant tumors affecting women is breast cancer. This tumor develops and occurs due to several internal and external factors. Resistance remains a key challenge in modern breast cancer therapy. Novel 1-substituted isatin-5-sulfonamides with antiproliferative effects based on isatin-core-containing antitumor compounds were synthesized in three stages via alkylation using benzyl chlorides. The study focuses on the synergistic effect of the obtained 1-substituted isatin-5-sulfonamides, exhibiting pro-apoptotic activity and is combined with heat shock protein 90 (HSP90) and protein kinase B (AKT) selective inhibitors in breast cancer cell lines, which are sensitive and resistant to antiestrogens.To create resistance, 4-hydroxytamoxifen (HT) was applied to create a resistant cell subline (MCF7/HT), achieving a resistance index of 2. MCF7/p53-LUC cell subline was obtained through transfection using the p53-responsive luciferase reporter plasmid. The lead compound LCTA-3344, exhibited the most significant antiproliferative effect, with a lower half-maximal inhibitory concentration (IC(50) ) in MCF7/HT (1.4±0.1 μМ) compared to MCF7 (2.6±0.3 μМ). Synergistic effects were observed when Combining the apoptosis inducer LCTA-3344 and AKT Inhibitor IV in both MCF7 and MCF7/HT, demonstrating the combination index (CI) values of 0.8 and 0.4, respectively (indicating higher activity). Apoptosis inducer LCTA-3344 combined with AKT Inhibitor X and HSP90 inhibitor did not show such significant activity with a minimal CI value of 0.9. Notably,Compound LCTA-3344 did not enhance luciferase activity in the MCF7/p53-LUC cell subline, while chemotherapeutic agent doxorubicin has been determined to be its potent inducer. In conclusion, apoptosis inducer LCTA-3344 was 1.9-fold more active toward MCF7/HT in comparison to the parental cell line. Compound LCTA-3344 together with AKT Inhibitor IV was the most active drug combination on the MCF7/HT subline, with a CI of 0.4. Compound LCTA-3344 induced apoptosis through a p53-independent mechanism, which holds promise as a novel therapy for hormone-resistant breast cancers. AKT Inhibitor IV caused apoptosis of MCF7 cells to a greater extent than compound LCTA-3344, and their combination resulted in a synergistic effect.

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