Liu Tian, Long Tian, Li Haosen
Department of Stomatology, The Central Hospital of Wuhan, Wuhan, Hubei 430000, P.R. China.
Department of Stomatology Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430000, P.R. China.
Exp Ther Med. 2021 Mar;21(3):202. doi: 10.3892/etm.2021.9635. Epub 2021 Jan 11.
Oral squamous cell carcinoma (OSCC) is the most common cancer of the oral cavity. Curcumin (Cur), a naturally derived compound, is reported to have broad-spectrum anticancer activity and is considered as an effective nuclear factor-κB (NF-κB) inhibitor. The present study aimed to clarify the detailed molecular mechanism though which Cur regulates NF-κB pathway activity in OSCC. The viability of HSC3 and CAL33 cells following treatment with Cur was determined using a Cell Counting Kit-8 assay. The protein and mRNA expression of specificity protein 1 (Sp1), p65 and heat shock factor 1 (HSF1) was determined by western blotting and reverse transcription-quantitative PCR analysis, respectively. The NF-κB activity was measured by Dual-Luciferase reporter assay. Short hairpin RNA targeting Sp1 or control RNA was transfected into HSC3 cells using X-treme GENE HP DNA Transfection System. Colony formation assays were performed using crystal violet staining. The results demonstrated that Cur significantly inhibited the viability and colony formation ability of HSC3 and CAL33 cells. In addition, Cur decreased the expression of Sp1, p65 and HSF1 by suppressing their transcription levels. Cur decreased NF-κB activity in OSCC cells, and Sp1 downregulation enhanced the effect of Cur. The findings from the present study suggested that Cur may inhibit the proliferation of OSCC cells via a Sp1/NF-κB-dependent mechanism.
口腔鳞状细胞癌（OSCC）是口腔中最常见的癌症。姜黄素（Cur）是一种天然衍生的化合物，据报道具有广谱抗癌活性，被认为是一种有效的核因子-κB（NF-κB）抑制剂。本研究旨在阐明Cur调节OSCC中NF-κB信号通路活性的详细分子机制。使用细胞计数试剂盒-8检测法测定Cur处理后HSC3和CAL33细胞的活力。分别通过蛋白质印迹法和逆转录-定量PCR分析测定特异性蛋白1（Sp1）、p65和热休克因子1（HSF1）的蛋白质和mRNA表达。通过双荧光素酶报告基因检测法测量NF-κB活性。使用X-treme GENE HP DNA转染系统将靶向Sp1的短发夹RNA或对照RNA转染到HSC3细胞中。使用结晶紫染色进行集落形成试验。结果表明，Cur显著抑制了HSC3和CAL33细胞的活力和集落形成能力。此外，Cur通过抑制Sp1、p65和HSF1的转录水平来降低它们的表达。Cur降低了OSCC细胞中的NF-κB活性，而Sp1的下调增强了Cur的作用。本研究结果表明，Cur可能通过Sp1/NF-κB依赖性机制抑制OSCC细胞的增殖。
