非小细胞癌(NSCLC)是世界上最常见的癌症,但其有效的治疗方法有限。Tilianin和舒芬太尼缓解了各种人类肿瘤。本研究旨在阐明替立宁和舒芬太尼在非小细胞肺癌中的作用及机制。Tilianin和舒芬太尼对NSCLC细胞活力的作用,凋亡,线粒体功能障碍,使用细胞计数试剂盒-8测定法检查体外免疫,流式细胞术,活性氧水平分析,CD8+T细胞百分比分析,蛋白质印迹,和酶联免疫吸附测定,分别。用Westernblot方法评估替立宁和舒芬太尼在非小细胞肺癌中调节的分子机制,和免疫荧光分析。同时,替立宁和舒芬太尼在非小细胞肺癌肿瘤生长中的作用,凋亡,并通过建立肿瘤异种移植小鼠模型测定体内免疫力,免疫组织化学,和蛋白质印迹分析。当舒芬太尼浓度接近2nM时,NSCLC细胞活力抑制率为50%。A549细胞的IC50为2.36nM,H1299细胞的IC50为2.18nM。Tilianin对A549细胞的IC50为38.7μM,Tilianin对H1299细胞的IC50为44.6μM。功能上,0.5nM舒芬太尼和10μMTilianin以剂量依赖性方式降低NSCLC细胞(A549和H1299)活力。此外,0.5nM舒芬太尼和10μMTilianin增强NSCLC细胞凋亡,然而,这种影响在Tilianin和舒芬太尼联合使用后得到加强.此外,0.5nM舒芬太尼和10μMTilianin抑制NSCLC细胞线粒体功能障碍和免疫,这些影响在Tilianin和舒芬太尼联合用药后得到增强.机械上,0.5nM舒芬太尼和10μMTilianin抑制NSCLC细胞中的NF-κB通路,而这种抑制在联合使用Tilianin和舒芬太尼后得到加强。体内实验数据进一步阐明,1µg/kg舒芬太尼和10mg/kgTilianin降低了NSCLC的生长,豁免权,和NF-κB通路相关蛋白水平,然而,在Tilianin和舒芬太尼联合用药后,这些趋势得到了增强.Tilianin增强舒芬太尼对NSCLC的抗肿瘤作用。
Non-small cell cancer (NSCLC) is the most common cancer in the world, but its effective therapeutic methods are limited. Tilianin and sufentanil alleviate various human tumors. This research aimed to clarify the functions and mechanisms of Tilianin and sufentanil in NSCLC. The functions of Tilianin and sufentanil on NSCLC cell viability, apoptosis, mitochondrial dysfunction, and immunity in vitro were examined using Cell Counting Kit-8 assay, flow cytometry, reactive oxygen species level analysis, CD8+ T cell percentage analysis, Western blot, and enzyme-linked immunosorbent assay, respectively. The molecular mechanism regulated by Tilianin and sufentanil in NSCLC was assessed using Western blot, and immunofluorescence assays. Meanwhile, the roles of Tilianin and sufentanil in NSCLC tumor growth, apoptosis, and immunity in vivo were determined by establishing a tumor xenograft mouse model, immunohistochemistry, and Western blot assays. When sufentanil concentration was proximity 2 nM, the inhibition rate of NSCLC cell viability was 50%. The IC50 for A549 cells was 2.36 nM, and the IC50 for H1299 cells was 2.18 nM. The IC50 of Tilianin for A549 cells was 38.7 μM, and the IC50 of Tilianin for H1299 cells was 44.6 μM. Functionally, 0.5 nM sufentanil and 10 μM Tilianin reduced NSCLC cell (A549 and H1299) viability in a dose-dependent manner. Also, 0.5 nM sufentanil and 10 μM Tilianin enhanced NSCLC cell apoptosis, yet this impact was strengthened after a combination of Tilianin and Sufentanil. Furthermore, 0.5 nM sufentanil and 10 μM Tilianin repressed NSCLC cell mitochondrial dysfunction and immunity, and these impacts were enhanced after a combination of Tilianin and Sufentanil. Mechanistically, 0.5 nM sufentanil and 10 μM Tilianin repressed the NF-κB pathway in NSCLC cells, while this repression was strengthened after a combination of Tilianin and Sufentanil. In vivo experimental data further clarified that 1 µg/kg sufentanil and 10 mg/kg Tilianin reduced NSCLC growth, immunity, and NF-κB pathway-related protein levels, yet these trends were enhanced after a combination of Tilianin and Sufentanil. Tilianin strengthened the antitumor effect of sufentanil in NSCLC.