肌醇 1,4,5-三磷酸受体 (IP3R) 是一种关键的钙离子 (Ca2+) 调节剂，在乳腺癌 (BC) 代谢中发挥着至关重要的作用。 BC 细胞中 IP3R 失调可导致异常生长或细胞死亡。雌二醇会增加 BC 中 IP3R 3 型 (IP3R3) 的水平，促进细胞增殖和代谢变化，包括增强丙酮酸脱氢酶 (PDH) 活性，当丙酮酸脱氢酶 (PDH) 活性降低时，会导致细胞凋亡。该研究沉默 IP3R3 以评估其对 PDH 的影响。该研究使用 IP3R3 小干扰 RNA (siRNA) 靶向密歇根癌症基金会-7 (MCF-7) 和 MDA-MB-231 细胞系。通过流式细胞术证实转染成功。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑 (MTT) 和实时定量聚合酶链式反应 (PCR) 测定法评估细胞活力和基因沉默。通过western blotting和PDH活性测定分析蛋白表达和细胞活性。用IP3R3 siRNA转染MCF-7和MDA-MB-231细胞，转染率达到65%，且无明显毒性。 IP3R3基因沉默有效降低了两种细胞系中IP3R3信使RNA (mRNA)和蛋白质水平，导致PDH酶活性降低，尤其是在MDA-MB-231细胞中。该研究强调了IP3R3基因高度沉默与PDH活性降低之间的联系，与雌激素依赖性 (MCF-7) 细胞系相比，雌激素非依赖性 (MDA-MB-231) 细胞系中 IP3R3 表达更高。这表明通过调节 PDH 活性对 BC 代谢和肿瘤生长产生潜在影响。版权所有：© 2024 Advanced Biomedical Research。

Inositol 1,4,5-trisphosphate receptor (IP3R), a critical calcium ion (Ca2+) regulator, plays a vital role in breast cancer (BC) metabolism. Dysregulated IP3R in BC cells can drive abnormal growth or cell death. Estradiol increases IP3R type 3 (IP3R3) levels in BC, promoting cell proliferation and metabolic changes, including enhanced pyruvate dehydrogenase (PDH) activity, which, when reduced, leads to cell apoptosis. The study silenced IP3R3 to assess its impact on PDH.The study used IP3R3 small interfering RNA (siRNA) to target Michigan Cancer Foundation-7 (MCF-7) and MDA-MB-231 cell lines. Transfection success was confirmed by flow cytometry. Cell viability and gene silencing were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and real-time quantitative polymerase chain reaction (PCR) assays. Protein expression and cellular activity were analyzed through western blotting and PDH activity measurement.Transfecting MCF-7 and MDA-MB-231 cells with IP3R3 siRNA achieved a 65% transfection rate without significant toxicity. IP3R3 gene silencing effectively reduced IP3R3 messenger RNA (mRNA) and protein levels in both cell lines, leading to decreased PDH enzyme activity, especially in MDA-MB-231 cells.The study highlights a link between high IP3R3 gene silencing and reduced PDH activity, with higher IP3R3 expression in estrogen-independent (MDA-MB-231) compared to estrogen-dependent (MCF-7) cell lines. This suggests a potential impact on BC metabolism and tumor growth via regulation of PDH activity.Copyright: © 2024 Advanced Biomedical Research.