A novel lupene derivative from Thymus capitatus possesses an apoptosis-inducing effect via Let-7 miRNA/Cyclin D1/VEGF cascade in the A549 cell line

Sep 24, 2026

Non-small-cell lung carcinoma (NSCLC) accounts for about 85% of lung cancers, and the study isolates and evaluates a novel lupene derivative, acetoxy-lup-5(6), 20(29)-diene (ALUP), alongside lupeol (LUP) and betulinic acid (BA) from Thymus capitatus. Compounds were purified by chromatographic methods, identified via physicochemical properties and 1D/2D NMR, and tested using MTT and annexin V/propidium iodide assays. Results show G2/M cell-cycle arrest and apoptosis induction in A549 cells through a Let-7 miRNA/Cyclin D1/VEGF pathway, with upregulated Let-7, reduced miRNA-21, and altered Bax, CASP-8, CD95, Bcl-2, KRAS, VEGF, and Cyclin D1 expression.

LanguageEnglish

CategoryHealthcare

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Pages12

Reading Time30 min

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Aborehab et al. BMC Complementary Medicine and Therapies (2023) 23:365 [https://doi.org/10.1186/s12906-023-04201-7](https://doi.org/10.1186/s12906-023-04201-7) BMC Complementary Medicine and Therapies RESEARCH Open Access A novel lupene derivative from Thymus capitatus possesses an apoptosis-inducing effect via Let-7 miRNA/Cyclin D1/VEGF cascade in the A549 cell line Nora M. Aborehab 1, Maha M. Salama2,3 and Shahira M. Ezzat2,4* Abstract 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 nonsmall 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 acetoxylup-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 bea potential therapeutic agent for lung cancer. Keywords Thymus capitatus L., Lupeol derivatives, A549 human non-small cell lung cancer cell line, miRNA-21, Let-7 miRNA *Correspondence: Shahira M. Ezzat [smelkomy@msa.edu.eg](smelkomy@msa.edu.eg); [shahira.ezzat@pharma.cu.edu.eg](shahira.ezzat@pharma.cu.edu.eg) Full list of author information is available at the end of the article © The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ()[. The Creative Commons Public Domain Dedication waiver (](http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ()[http://creativecom](http://creativecom)[mons.org/publicdomain/zero/1.0/](mons.org/publicdomain/zero/1.0/)) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Aborehab et al. BMC Complementary Medicine and Therapies (2023) 23:365 Page 2

FAQs

What compounds were isolated from Thymus capitatus in this study?

The study isolated a novel lupene derivative, acetoxy-lup-5(6), 20(29)-diene (ALUP), and two known triterpenes: lupeol (LUP) and betulinic acid (BA).

How were the isolated compounds identified?

Identification used physicochemical properties and spectral 1D and 2D NMR analysis.

What cellular effects did ALUP, LUP, and BA produce in A549 cells?

They caused G2/M phase cell-cycle arrest and increased apoptosis, linked to changes in Let-7 miRNA/Cyclin D1/VEGF-related molecular expression.

Sep 24, 2026

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