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All Journal Eureka Herba Indonesia Sriwijaya Journal of Forensic and Medicolegal
Leonardo Simanjuntak
Department of Obstetrics and Gynecology, CMHC Research Center, Palembang, Indonesia
Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Law, Crime, Criminology & Criminal Justice Medicine & Pharmacology

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Apoptotic Induction and Cell-Cycle Arrest in Triple-Negative Breast Cancer Cells by Annona muricata Acetogenin Fractions: An In Silico and In Vitro Study Lestini Wulansari; Leonardo Simanjuntak; Vania Delma
Eureka Herba Indonesia Vol. 6 No. 2 (2025): Eureka Herba Indonesia
Publisher : HM Publisher

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37275/ehi.v6i2.142

Abstract

Triple-negative breast cancer (TNBC) is an aggressive molecular subtype that lacks oestrogen, progesterone and HER2 receptors and retains cytotoxic chemotherapy as its principal systemic option, creating a rationale for adjunctive phytotherapy. Annona muricata L. (Annonaceae), known in Indonesia as sirsak, contains Annonaceous acetogenins with well-documented mitochondrial and apoptotic activity. This study integrated molecular docking with in vitro pharmacology to characterise the anticancer effects of an HPLC-standardised acetogenin-enriched fraction (AEF) from Indonesian A. muricata leaves on TNBC cells. Annonacin, bullatacin, squamocin and muricatacin were docked against EGFR, Bcl-2, CDK2, caspase-3 and topoisomerase II-α (AutoDock Vina); AEF was tested on MDA-MB-231 and MDA-MB-468 TNBC cells with MCF-10A non-tumorigenic controls (n = 6 independent biological replicates per arm). Outcomes included viability (MTT), apoptosis (Annexin-V/PI), cell-cycle distribution (propidium iodide), caspase-3/7 luminescence, Western blotting (Bcl-2, Bax, cleaved caspase-3, cyclin D1, p21) and mitochondrial ΔΨm/ROS. Bullatacin showed the strongest binding to Bcl-2 (ΔG = −9.6 kcal/mol) and caspase-3 (ΔG = −8.9 kcal/mol). AEF inhibited MDA-MB-231 viability with IC50 12.4 µg/mL (95% CI 10.8–14.1) and yielded a selectivity index of 4.20 over MCF-10A. Apoptotic cells increased 4.62-fold, the G1 fraction rose from 41.2% to 64.8% (p < 0.001), caspase-3/7 activity rose 3.81-fold and the Bcl-2/Bax ratio decreased by 61%. In conclusion, bullatacin-rich Annona muricata acetogenins selectively induce intrinsic apoptosis and G1 arrest in TNBC cells, supporting their further translational development as Indonesian phytotherapeutic leads for hormone-refractory breast cancer.
Reading the Epigenetic Clock: A Comparative Analysis of DNA Methylation Markers for Age Estimation in Semen, Saliva, and Touch DNA Febria Suryani; Bryan Helsey; Leonardo Simanjuntak; Karina Chandra; Mustafa Mahmud; Lisha Sandrina; Ahmad Erza
Sriwijaya Journal of Forensic and Medicolegal Vol. 3 No. 1 (2025): Sriwijaya Journal of Forensic and Medicolegal
Publisher : Phlox Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.59345/sjfm.v3i1.233

Abstract

Background. Predicting an individual's age from biological evidence is a significant advance in forensic intelligence. DNA methylation, a stable epigenetic mark, provides the molecular basis for epigenetic clocks, but their operational reliability requires validation across diverse sample types and populations, particularly for low-template touch DNA. Objective. To develop and validate body-fluid-specific age prediction models from a curated five-CpG panel in semen, saliva and touch DNA, and to compare their accuracy in an Indonesian population. Methods. Following approval from the Ethical Committee of CMHC Indonesia (No. 128/EC/CMHC/2023), 150 healthy Indonesian male volunteers aged 18–65 provided semen, saliva and high-yield standardized touch DNA. Methylation at five CpG sites (ELOVL2, FHL2, TRIM59, KCNQ1DN, C1orf132) was quantified by controlled pyrosequencing after bisulfite conversion with efficiency controls. Body-fluid-specific models were built by multiple linear regression and validated by 10-fold cross-validation. Results. The semen and saliva models were highly accurate, with mean absolute deviations of 3.19 years (R²=0.94) and 3.55 years (R²=0.92). The touch DNA model was less precise but still informative (MAD 5.49 years, R²=0.85). All models satisfied the assumptions of linear regression, with variance inflation factors below 2.5, and 95% prediction intervals were narrowest for semen. Conclusion. The panel is validated for age prediction in a Southeast Asian population. The semen and saliva models are accurate enough for consideration in casework, while the touch DNA model, interpreted cautiously, can generate investigative leads from trace evidence. The findings underline the importance of tissue-specific modeling and provide a methodological blueprint for responsible forensic age estimation.
Co-Authors Ahmad Erza Bryan Helsey Febria Suryani Karina Chandra Lestini Wulansari Lisha Sandrina Mustafa Mahmud Vania Delma