INTRODUCTION:

Cancer remains one of the leading causes of mortality worldwide, with breast and cervical cancer^1,2 being among the most prevalent malignancies affecting women³. Despite advancements in cancer therapy, including chemotherapy, targeted therapy, and immunotherapy⁴. Significant challenges persist, particularly in terms of drug selectivity and adverse side effects⁵. Chemotherapeutic agents, such as paclitaxel, doxorubicin, and cisplatin, are widely used but often exhibit non-selective cytotoxicity, affecting both cancerous and normal cells^6–8. This non-specific action leads to severe side effects, including myelosuppression, neuropathy, cardiotoxicity, and gastrointestinal disturbances, which significantly impact patient quality of life and treatment adherence^9–11.

To improve therapeutic outcomes and minimize toxicity, developing selective anticancer agents has become a major focus in oncology research¹². The Selectivity Index (SI) is a crucial parameter for evaluating the therapeutic potential of cytotoxic compounds^13,14. SI is defined as the ratio of cytotoxicity in normal cells (IC₅₀ normal cells) to that in cancer cells (IC₅₀ cancer cells)¹⁵, with higher SI values (>1) indicating better selectivity towards cancer cells over normal cells¹⁶. For instance, clinically used chemotherapeutic agents often demonstrate varying degrees of selectivity⁴. Paclitaxel, a well-established microtubule-stabilizing agent^17,18, exhibits moderate selectivity, which can lead to significant neuropathy¹⁹, pain²⁰, and neutropenia^21,22. In contrast, targeted therapies such as trastuzumab (HER-2 inhibitor) show higher selectivity²³, reducing off-target effects but remaining ineffective in patients without HER-2 overexpression^24,25.

The Taxus species, particularly T. sumatrana (Miq.) de Laub., has attracted attention due to its potential anticancer properties, which arise from its bioactive compounds, including taxanes, which are key components of chemotherapeutic drugs²⁶. Previous studies have reported that T. sumatrana extracts exhibit significant cytotoxic effects on various cancer cell lines, with concentrations below 20µg/mL significantly inhibiting the growth of breast (T47D, MCF-7/HER-2) and cervical (HeLa) cancer cells²⁷. However, the selectivity of these extracts towards cancer cells over normal cells remains underexplored, and further investigation is needed to assess their therapeutic potential²⁸.

To address this gap, this study focused on assessing the selectivity of T. sumatrana extracts derived from different plant parts (bark, leaves, and shoots) by comparing their cytotoxic effects on breast cancer (T47D, MCF-7/HER-2) and cervical cancer (HeLa) cell lines with those on normal Vero cells. By determining the Selectivity Index (SI) of each extract, this research aimed to identify plant-derived compounds with high anticancer selectivity, thereby minimizing toxicity in normal tissues. The findings are expected to provide valuable insights into the potential of T. sumatrana as a source of selective anticancer agents, contributing to the development of safer and more effective herbal-based cancer therapies.

MATERIALS AND METHODS:

Plant Material and Extract Preparation:

Taxus sumatrana (Miq.) de Laub. was collected from Mount Singgalang in West Sumatra, Indonesia. The botanist identified the plant specimen, and a voucher specimen (No DEP012024) has been deposited at the ANDA Herbarium, Universitas Andalas, Padang, Indonesia. Plant materials (bark, leaves, shoots) were air-dried for two weeks and ground into powder. The powdered bark was macerated with 70% ethanol for 72 hours while stirring. The extract was filtered and concentrated using a rotary evaporator below 40°C. The yield of the extract was calculated according to Eq. (1).

Weight of the extract

% Yield = --------------------------------- x 100 %

Weight of the macerated sample

Eq. (1)

The final crude extract was stored in a sealed glass bottle wrapped in aluminum foil at 2–8°C until use²⁹.

Phytochemical Screening:

The phytochemical composition of T. sumatrana bark, leaf, and shoot extracts was analyzed to identify the presence of alkaloids, flavonoids, phenolics, saponins, terpenoids, and steroids. Standard qualitative phytochemical tests were performed following established protocols to confirm the presence of these bioactive compounds³⁰. Each test was conducted using specific reagents and observation criteria to identify the presence of targeted secondary metabolites.

Paclitaxel Identification:

The presence of paclitaxel in T. sumatrana extracts was confirmed using High-Performance Liquid Chromatography (HPLC) following the specifications outlined in the Indonesian Pharmacopoeia³¹. The analysis used a C18 column with a mobile phase composed of acetonitrile and water. Detection was carried out at a specific wavelength, and the paclitaxel content was determined by comparing the retention time and peak characteristics with those of a reference standard (USP Reference Standard).

Cell Culture:

The cytotoxicity assays utilized the following human cell lines: breast cancer cell lines T47D and MCF-7/HER-2, the cervical cancer cell line HeLa, and normal Vero cells. The inclusion of normal Vero cells (African green monkey kidney cells) served as a control to assess the selectivity of the extracts between cancerous and normal cells. Cells were sourced from ATCC. They were cultured in Dulbecco’s Modified Eagle Medium (DMEM) or RPMI-1640, both supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin, and incubated at 37°C in a humidified atmosphere with 5% CO₂.

Cytotoxicity Assay (MTT Assay):

The cytotoxic activity (inhibition) of T. sumatrana extracts was evaluated using the MTT assay^27,32. Cells were plated in 96-well plates at a density of 6,000 cells per well and incubated overnight for adhesion. After 24 h, the cells were treated with various concentrations of bark, leaf, and shoot extracts (1–1000 µg/mL) for 24/48 h. Following treatment, 10 µL of MTT solution (5 mg/mL) was added to each well and incubated for 4 hours at 37°C. The formazan crystals were dissolved in DMSO, and absorbance was measured at 570 nm using a microplate reader. % Inhibition or cytotoxicity activity (%) was calculated relative to the untreated control according to Eq. (2).

The absorbance of trated cells

% Inhibition = 1- ( --------------------------------- ) x100

The absorbance of control cells

………..Eq. (2)

The IC₅₀ values (concentration inhibiting 50% cell growth) were determined using GraphPad Prism.

Selectivity Index (SI):

The Selectivity Index (SI) was calculated as Eq. (3)³³:

IC10 in normal cells

SI= -------------------------- ………..Eq. (3)

IC10 in cancer cells

An SI value greater than 1 indicates selective cytotoxicity towards cancer cells³³.

Statistical Analysis:

All experiments were conducted in triplicate, and results were presented as mean ± standard deviation (SD). Statistical differences between Vero and cancer cell groups were assessed using one-way ANOVA, followed by Tukey’s post hoc test. All statistical analyses were conducted using GraphPad Prism, with a p-value of <0.05 considered statistically significant.

RESULTS:

The extraction yields of the bark, leaf, and shoot extracts of T. sumatrana ranged from 50% to 80%, calculated based on the dry weight of each macerated sample. These comparatively high yields suggest an efficient recovery of soluble constituents from each plant part using 70% ethanol as the extraction solvent. Phytochemical screening of T. sumatrana bark, leaf, and shoot extracts showed the presence of flavonoids, phenolics, saponins, terpenoids, and steroids in all three samples. Alkaloids were not detected in any of the extracts (Table 1).

The presence of paclitaxel in the bark, leaf, and shoot extracts of T. sumatrana was analyzed using high-performance liquid chromatography (HPLC). The paclitaxel standard was identified at a retention time of approximately 19.9 minutes. Paclitaxel was detected in all three extracts at varying concentrations, with the highest peak intensity observed in the bark extract (Figure 1 A-D).

Table 1. Qualitative phytochemical screening of T. sumatrana bark, leaf, and shoot extracts

Samples	Phytochemical screening result
Samples	Alkaloids	Flavonoids	Phenolics	Saponins	Terpenoids	Steroids
Bark extract	-	+	+	+	+	+
Leaf extract	-	+	+	+	+	+
Shoot extract	-	+	+	+	+	+

+ detected - not detected

Figure 1. HPLC chromatograms of T. sumatrana extracts and the paclitaxel standard at 254 nm: (A) bark extract, (B) leaf extract, (C) shoot extract, and (D) paclitaxel standard. The paclitaxel peak appeared at approximately 19.9min in all extracts, with the highest intensity in the bark extract. The analysis was performed using a C18 column, with a flow rate of 1 mL/min, a column temperature of 30°C, and a mobile phase composed of acetonitrile: water (35:65)

The cytotoxicity of T. sumatrana extracts was assessed in normal cells and various cancer cell lines (T47D, MCF-7/HER-2, and HeLa) using the MTT assay. The cytotoxic effects depended on the concentration, with the most significant inhibition observed at 100µg/mL across all extracts. The inhibition of Vero cell growth was consistently lower than that of cancer cells (Figure 2).

Figure 2. Comparison of growth inhibition (%) of T. sumatrana bark, leaf, and shoot extracts on Vero and cancer cell lines (T47D, MCF-7/HER-2, and HeLa) at various concentrations. Data are presented as mean±SD from three independent experiments.

The cytotoxic potency of T. sumatrana bark, leaf, and shoot extracts was evaluated based on their IC₅₀ values in Vero, T47D, MCF-7/HER2, and HeLa cell lines. The IC₅₀ values varied across cell types, with higher values observed in Vero cells compared to the cancer cell lines (Table 2). The IC₅₀ values in Vero cells were 40.10 µg/mL for the bark extract, 61.14µg/mL for the leaf extract, and 38.63µg/mL for the shoot extract. In contrast, the IC₅₀ values in T47D, MCF-7/HER2, and HeLa cells were approximately 2 to 21 times lower, consistently across all three extracts. Statistical analysis using one-way ANOVA demonstrated significant differences in IC₅₀values between cancer and normal cells within each extract group (Figure 3).

Table 2. IC₅₀ value of T. sumatrana bark, leaf, and shoot extract in Vero, T47D, MCF-7/HER-2, and HeLa cells.

Cell lines	IC₅₀(µg/ml)
Cell lines	Bark extract	Leaf extract	Shoot extract
Vero	40.10 ± 5.89	61.14 ± 15.50	38.63 ± 1.18
T47D	5.80 ± 0.79	4.86 ± 2.20	4.10 ± 3.07
MCF-7/HER-2	7.46 ± 1.74	10.60 ± 0.27	13.74 ± 1.46
HeLa	8.94 ± 1.61	5.93 ± 2.86	4.078 ± 1.27

The IC₅₀ values are presented as mean ± SD in µg/mL. Each value was derived from three independent experiments conducted in triplicate.

Figure 3. Comparison of IC₅₀ values of T. sumatrana bark, leaf, and shoot extracts on Vero, T47D, MCF-7/HER2, and HeLa cells. Data are presented as mean ± SD from three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. Significant differences were observed between Vero cells and each cancer cell line for all extracts (p<0.05). ***p< 0.0005, ****p<0.0001

The Selectivity Index (SI) values of T. sumatrana bark, leaf, and shoot extracts in Vero cells compared to T47D, MCF-7/HER2, and HeLa cancer cell lines are presented in Figure 4. All extracts showed SI values greater than 1. The highest SI value was observed for the shoot extract in T47D cells (21.92), and the lowest SI value was for the shoot extract in HeLa cells (2.84). Overall, the three extracts exhibited the greatest selectivity toward T47D cells (red bar), followed by HeLa (purple bar) and MCF-7/HER2 cells (green bar).

Figure 4. Selectivity Index (SI) values of extracts from T. sumatrana in Vero cells compared to cancer cell lines (T47D, MCF-7/HER2, and HeLa). All extracts demonstrate SI values greater than 1 (dashed line), indicating selective activity against cancerous cells. Data are presented as mean±SD from three independent experiments.

DISCUSSION:

Selectivity has become a key consideration in cancer therapy, driven by the clinical reality that many currently available chemotherapeutic agents, although effective, are often limited by their non-selective toxicity to normal cells. This underscores the urgent need for therapeutic agents that not only effectively target cancer cells but also minimize harm to healthy tissues. In this context, the present study offers a comparative analysis of the cytotoxic selectivity profile of ethanol extracts from three parts of T. sumatrana (bark, leaves, and shoots) by assessing their effects on normal and cancerous cell lines.

All three extracts showed cytotoxic activity against T47D, MCF-7/HER2, and HeLa cancer cell lines, with IC₅₀ values below 15µg/mL for all extracts. These values fall within the threshold generally considered promising for plant-derived anticancer agents, as established by the U.S. National Cancer Institute (NCI), which considers extracts with IC₅₀ values below 30µg/mL to have significant in vitro cytotoxic potential³⁴. When compared to previously reported cytotoxicity data of natural products, such as guar saponin extract (IC₅₀12.4µg/mL)³⁵ and soybean saponin extract (IC₅₀ 6.875µg/mL) against MCF-7 cells³⁶, or alcoholic extract of Aristolochia indica (IC₅₀ 28.56µg/mL) on HT-29 colon cancer cells³⁷, the results of this study demonstrate comparable potency.

The extracts displayed lower cytotoxicity toward normal Vero cells, with IC₅₀ values more than twice those observed in cancer cells. This pattern suggests a tendency for the extracts to act more strongly on malignant cells than on normal cells, which is a desirable characteristic in candidate anticancer agents. Similar anticancer potential has also been reported in other Taxus species³⁸, such as T. baccata³⁹ and T. brevifolia⁴⁰. The calculated Selectivity Index (SI) values for all extracts exceeded 1, indicating differential cytotoxic effects between cancer and normal cells^16,33. Among the three, the shoot extract demonstrated the highest SI, followed by the leaf and bark extracts.

The SI is an important early indicator in the screening of both synthetic and natural anticancer agents, providing initial insights into the balance between cytotoxic potency and safety⁴¹. Previous studies on natural products have shown a wide range of SI values. For example, Taxus baccata extracted from Spanish medicinal plants exhibited an SI of 157.3±110.6 against A549 lung cancer cells⁴² and. Methanolic leaf extract of Moringa oleifera showed an SI of 9.5 against MCF-7 cells⁴³, while Garcinia cowa ethanol extract had an SI of 2.55⁴⁴. A study of 15 traditional Mexican medicinal plants reported SI values ranging from 1 to >20 in lymphoma cell lines⁴⁵. These findings highlight the variability in selectivity among natural compounds and emphasize the necessity of comprehensive SI evaluations in phytopharmacological research.

In our study, the SI values of T. sumatrana extracts fell within a comparable range and, in some cases, even exceeded those of well-established chemotherapeutic agents. Doxorubicin and cisplatin have been reported to show SI values of 1.31 and 2.11, respectively, against HeLa cells, whereas paclitaxel demonstrated an SI of 0.94⁴⁶. Against other cancer types, doxorubicin and 5-fluorouracil (5-FU) showed SI values of 22.53 and 1.93 for MCF-7 and HT-29 cells, respectively⁴⁷. In addition, a recent study reported that 4-methyl-3-benzoyl allylthiourea, a novel anticancer candidate, exhibited SI values of 2 for T47D cells and 4 for MCF-7/HER2 cells⁴⁸. Although such comparisons should be interpreted cautiously due to differences in cell types, extract composition, and assay conditions, our findings provide initial context for the potential selectivity of T. sumatrana extracts.

These observations are particularly relevant when viewed in light of the limitations of current chemotherapeutics. While paclitaxel is clinically effective in nasopharyngeal carcinoma cells⁴⁹, human ovarian cancer cells⁵⁰, oral cancer cells⁵¹, and lung cancer cells⁵², it is associated with systemic toxicity, such as peripheral neuropathy⁵³, neutropenia⁵⁴, and cardiac disorder⁵⁵. These effects are consistent with its low in vitro SI value (<1) observed in a previous in vitro study⁴⁷. Therefore, the relatively higher SI value of T. sumatrana bark extract may indicate a more favorable therapeutic profile. However, this hypothesis requires further validation through in vivo studies and mechanistic investigations, similar to those conducted for several natural compounds such as ginseng and curcumin, which have shown promising results and are currently undergoing clinical evaluation⁵⁶.

One possible explanation for the observed SI values lies in the presence of multiple bioactive compounds within the extracts, which may act synergistically. HPLC analysis confirmed the presence of paclitaxel in all extracts, with the highest levels found in the bark. This is consistent with previous studies reporting paclitaxel concentrations of 0.473±0.031ppm in acetone extracts of the bark, while leaves were found to contain lower levels⁵⁷. Interestingly, despite having the lowest paclitaxel content, the shoot extract showed the highest SI, suggesting that other compounds such as flavonoids, phenolics, or terpenoids may also contribute to the selective cytotoxic effect.

Various studies have identified different secondary metabolites from distinct parts of Taxus plants. The bark has been reported to contain baccatin III, cephalomannine, and related compounds⁵⁸, while leaves and twigs produce unique compounds such as wallifoliol, taxuspine F, taxumairol⁵⁹, tasumatrols^60–65, taiwantaxins⁶⁶ and other chemical compounds^67,68. This diversity in metabolite profiles may underlie the selective cytotoxicity observed in this study.

It is important to acknowledge that T. sumatrana has been used in traditional medicine by various local communities. While such ethnomedical use provides a valuable foundation for scientific exploration, rigorous pharmacological and toxicological validation remains essential. This study represents one of the first systematic evaluations of the selective cytotoxicity of T. sumatrana extracts against breast and cervical cancer cells, underscoring the importance of integrating ethnopharmacological knowledge with experimental validation.

Although the SI values observed for the bark extract are promising, several limitations must be considered. This study was conducted in vitro, which may not reflect the complexity of metabolism, drug distribution, and long-term toxicity in whole organisms. Furthermore, the specific active compounds responsible for the observed selective effects have not yet been isolated or identified, limiting our understanding of the underlying cytotoxic mechanisms. Follow-up studies involving in vivo toxicity profiling, detailed phytochemical characterization, and mechanistic assays, such as apoptosis pathway analysis and cell cycle arrest studies, are necessary⁶⁹.

This study presents preliminary evidence of the selective cytotoxic potential of T. sumatrana bark extract. The extract demonstrated a more favorable SI profile compared to other plant parts and even compared to paclitaxel in some instances. However, further validation is required to confirm its therapeutic promise. The selective cytotoxic activity shown by the bark extract supports the scientific exploration of traditionally used medicinal plants, particularly those from biodiversity-rich regions that remain underrepresented in pharmacological research.

CONCLUSION:

This study demonstrated that ethanol extracts derived from the bark, leaves, and shoots of Taxus sumatrana exhibit selective cytotoxicity against breast (T47D, MCF-7/HER-2) and cervical (HeLa) cancer cell lines, while showing lower toxicity to normal Vero cells. All three extracts yielded Selectivity Index (SI) values greater than 1 across all cancer cell models, indicating favorable selectivity toward malignant cells. Notably, the shoot and leaf extracts displayed the highest SI values, especially in T47D cells, despite the bark extract containing the highest paclitaxel concentration. This suggests that other bioactive constituents may play a significant role in the observed selective effects.

These findings support the potential of T. sumatrana as a promising source of plant-derived anticancer agents and emphasize the relevance of plant part selection in natural product-based drug discovery. Further research is warranted to isolate the active compounds, explore their mechanisms of action, and evaluate their therapeutic potential through comprehensive in vivo studies.

CONFLICT OF INTEREST:

The authors declare that there are no conflicts of interest related to this study.

ACKNOWLEDGMENT:

The authors would like to acknowledge the Forestry Office of West Sumatra Province for their assistance in providing the plant samples used in this study.

FUNDING:

Universitas Andalas funded this Research through the International Collaboration Research (Penelitian Kolaborasi Luar Negeri, PKLN) scheme of the PKLN Top#200 cluster with contracts No. 415/UN16.19/PT.01.03/PKLN/2025.

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Received on 01.07.2025 Revised on 23.12.2025

Accepted on 24.03.2026 Published on 20.05.2026

Available online from May 25, 2026

Research J. Pharmacy and Technology. 2026;19(5):1969-1976.

DOI: 10.52711/0974-360X.2026.00282

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