Department of Biochemistry, Faculty of Pharmacy, October University for Modern Sciences and Arts (MSA), Giza, 12451, Egypt.
Department of Pharmacognosy, Faculty of Pharmacy, Cairo University, Kasr El-Ainy St., Cairo11562, Cairo, 11562, Egypt.
BMC Complement Med Ther. 2023 Oct 16;23(1):365. doi: 10.1186/s12906-023-04201-7.
Non-small-cell lung carcinoma (NSCLC) is a type of epithelial lung cancer accounting for about 85% of all lung cancers. In our research, a novel lupene derivative namely acetoxy-lup-5(6), 20(29)-diene (ALUP), as well as two known triterpenes; lupeol (LUP) and betulinic acid (BA) were isolated through the chromatographic purification of the 95% ethanolic extract of Thymus capitatus. Identification of the compounds was carried out by physicochemical properties as well as spectral 1D and 2D NMR analysis. The anti-cancer activity of the three triterpenes was assessed on non-small cell lung cancer cell line; A549 using MTT assay and cell cycle analysis using annexin V/propidium iodide. The molecular mechanism underlying anti-apoptotic effects was determined by analyzing Let-7 miRNA and miRNA-21 expression, the mRNA gene expression level of Bax, CASP-8, CD95, Bcl2, KRAS, VEGF, Cyclin D1 using qRT-PCR. Our results revealed that the three isolated compounds ALUP, LUP, and BA caused cell cycle arrest at the G2/M phase with an increase in the apoptosis which may be attributed to their significant effect on raising Bax, CASP-8, and CD95 and reducing the mRNA expression levels of Bcl-2, KRAS, VEGF, and Cyclin D1 compared to control cells. RT-PCR results showed that the ALUP, LUP, and BA significantly downregulated miRNA-21 expression. Meanwhile, the three compounds caused significant overexpression of Let-7 miRNA. This is the first report on the anti-cancer activity of acetoxy-lup-5(6), 20(29)-diene (ALUP) in reducing the proliferation and differentiation of the A549 cell line through inducing apoptosis. Finally, by targeting the Let-7 miRNA/Cyclin D1/VEGF cascade, acetoxy-lup-5(6), 20(29)-diene could be a potential therapeutic agent for lung cancer.
非小细胞肺癌（NSCLC）是一种上皮性肺癌，约占所有肺癌的 85%。在我们的研究中，通过对 Thymus capitatus 的 95%乙醇提取物进行色谱纯化，分离得到了一种新型的羽扇豆烷衍生物，即乙酰氧基-羽扇豆-5(6)，20(29)-二烯（ALUP），以及两种已知的三萜类化合物；羽扇豆醇（LUP）和白桦酸（BA）。通过物理化学性质和光谱 1D 和 2D NMR 分析鉴定了化合物的结构。采用 MTT 法测定了三种三萜类化合物对非小细胞肺癌细胞系 A549 的抗癌活性，并通过 Annexin V/PI 检测细胞周期分析。通过分析 Let-7 miRNA 和 miRNA-21 的表达、Bax、CASP-8、CD95、Bcl2、KRAS、VEGF、Cyclin D1 的 mRNA 基因表达水平，研究了抗凋亡作用的分子机制，采用 qRT-PCR。我们的研究结果表明，ALUP、LUP 和 BA 这三种分离化合物导致细胞周期停滞在 G2/M 期，细胞凋亡增加，这可能归因于它们显著提高了 Bax、CASP-8 和 CD95 的表达水平，降低了 Bcl-2、KRAS、VEGF 和 Cyclin D1 的 mRNA 表达水平，与对照组细胞相比。RT-PCR 结果表明，ALUP、LUP 和 BA 显著下调了 miRNA-21 的表达。同时，三种化合物显著上调了 Let-7 miRNA 的表达。这是首次报道乙酰氧基-羽扇豆-5(6)，20(29)-二烯（ALUP）通过诱导细胞凋亡，降低 A549 细胞系的增殖和分化，从而具有抗癌活性。最后，通过靶向 Let-7 miRNA/Cyclin D1/VEGF 级联，乙酰氧基-羽扇豆-5(6)，20(29)-二烯可能成为一种潜在的肺癌治疗药物。
