Article

In silico and In vitro Study of The Anticancer Potential of Essential

Oil from Sembung (

Blumea

balsamifera

) Leaves against HeLa Cervical

Cancer Cells

Rahmi Vika Ulia

1

, Toto Raharto

1

, Suryati

2

,

Adlis Santoni

2

,

Yoan De Nanda Herru

3

1

Department of Chemistry, Universitas Teknologi Nusantara, Bogor-16158, Indonesia

2

Department of Chemistry,

Faculty of Mathematics and Natural Sciences, Universitas Andalas, Padang-25163,

Indonesia

3

Department of Chemistry, Universitas Lancang Kuning, Pekanbaru- 28265, Indonesia

A

bstract

Blumea

balsamifera

(L.)

DC.

(sembung)

is

a

medicinal

plant

widely

recognized

for

its

diverse

pharmacological

properties,

including

cytotoxic

and

anticancer

activities.

However,

the

anticancer

potential of its leaf essential oil against cervical cancer remains poorly explored. This study aimed to

evaluate

the

anticancer

activity

of

B. balsamifera

leaf

essential

oil

against

HeLa

cervical

cancer

cells

using integrated in silico and in vitro approaches. Molecular docking analysis against the anti-apoptotic

Bcl-2 protein revealed favorable binding affinities for several major constituents, including camphor (-

5.745

kcal/mol),

caryophyllene

(-7.077

kcal/mol),

7-epi-silphiperfol-5-ene

(-7.311

kcal/mol),

and

γ-

eudesmol

(-7.083

kcal/mol),

indicating

potential

interactions

with

apoptosis-related

targets.

Furthermore, cytotoxic activity assessed by the MTT assay demonstrated an IC

₅₀

value of 32.90 µg/mL

against HeLa cells, indicating

moderate cytotoxic potency. These findings suggest that

B. balsamifera

leaf essential oil possesses promising anti-cervical cancer activity and may serve as a potential natural

source for the development of apoptosis-targeting anticancer agents.

Keywords:

Anticancer; Blumea balsamifera; molecular docking; MTT assay

*

Corresponding author

Email addresses: rahmivikaulia1@gmail.com (RV Ulia)

DOI: https://doi.org/

10.22437/chp.v10i1.49160

Received

October 10

th

2025;

Accepted

June 08

th

2026;

Available online

June 30

th

2026

(

https://creativecommons.org/licenses/by/4.0

)

171

Graphical Abstract

Introduction

The medicinal plant

Blumea balsamifera

(L.)

DC.,

commonly

known

as

sembung,

is

a

widely

distributed

shrub

in

Southeast

Asia

and

has

long

been

utilized

in

traditional

medicine

for

the

treatment

of

various

ailments,

including

fever,

wounds,

hemorrhoids,

diarrhea,

diabetes,

and

cancer-related

conditions

[1]

.

Phytochemical investigations have revealed

that

B.

balsamifera

contains

diverse

bioactive

constituents,

such

as

flavonoids,

terpenoids,

phenolic

compounds,

and

essential oils, which contribute to its broad

pharmacological

activities.

have

demonstrated

that

extracts

and

fractions

of

sembung

leaves

possess

significant

biological

properties,

including

antioxidant,

anti-inflammatory,

antimicrobial,

and

cytotoxic

activities.

Notably,

several

reports

have

shown

cytotoxic

and

antiproliferative

effects

against different cancer cell lines, including

A549 lung cancer cells, HL-60 leukemia cells,

and

HepG2

hepatocellular

carcinoma

cells

[2][3][4]

.

Thes

e

findings

highlight

the

potential

of

B.

balsamifera

as

a

promising

source

of

anticancer

agents

and

warrant

further investigation into its efficacy against

other clinically relevant malignancies.

Cervical

cancer

remains

a

major

global

health

burden

and

is

one

of

the

leading

causes

of

cancer-related

mortality

among

women,

particularly

in

low-

and

middle-

income

countries.

According

to

the

Global

Cancer

Observatory

(GLOBOCAN)

2022,

cervical

cancer

accounted

for

662,301

new

cases

and

348,874

deaths

worldwide,

ranking among the most prevalent cancers

affecting

women.

In

Indonesia,

cervical

cancer

is

the

third

most

frequently

diagnosed

cancer,

with

36,633

new

cases

reported

in

2022

[5]

.

Despite

advances

in

surgery,

radiotherapy,

chemotherapy,

and

targeted

therapies,

treatment

outcomes

remain limited by adverse side effects, drug

resistance,

tumor

recurrence,

and

high

treatment

costs.

Consequently,

the

identification

of

novel,

effective,

and

safer

anticancer

compounds

from

natural

resources has become an important area of

research in cancer drug discovery.

Among the various bioactive fractions of

B.

balsamifera

,

essential

oil

represents

a

particularly

attractive

source

of

pharmacologically active compounds due to

172

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

its

richness

in

volatile

terpenoids

and

oxygenated derivatives, many of which have

demonstrated

anticancer

properties.

However,

current

knowledge

regarding

the

anticancer

potential

of

B.

balsamifera

essential oil remains limited. Most previous

studies

have

focused

on

crude

extracts

or

solvent

fractions

and

have

primarily

evaluated their activity against non-cervical

cancer

models.

To

the

best

of

our

knowledge,

comprehensive

investigations

combining

molecular

docking

and

experimental

cytotoxicity

assessments

of

sembung

leaf

essential

oil

against

cervical

cancer cells are still scarce. This knowledge

gap

limits

our

understanding

of

the

molecular

mechanisms

underlying

its

potential anticancer effects.

The

B-cell

lymphoma-2

(Bcl-2)

protein

was

selected

as

the

molecular

target

in

this

study

because

it

plays

a

critical

role

in

regulating

apoptosis

and

is

frequently

overexpressed

in

cervical

cancer

cells,

including

HeLa

cells

[6][7]

.

Overexpression

of

Bcl-2

contributes

to

cancer

cell

survival

by

suppressing

programmed

cell

death,

thereby

promoting

tumor

progression

and

resistance

to

anticancer

therapy.

Inhibition

of

Bcl-2

has

therefore

emerged

as

an

attractive

therapeutic

strategy

for

inducing

apoptosis

in

cervical

cancer

cells.

Accordingly,

evaluating

the

interaction

of

essential

oil

constituents

with

Bcl-2

may

provide valuable insights into their potential

mechanism of action as anticancer agents.

Therefore,

this

study

aimed

to

investigate

the

anticancer

potential

of

B.

balsamifera

leaf

essential

oil

against

cervical

cancer

through

an

integrated

in silico

and

in

vitro

approach. Molecular docking was employed

to

predict

the

binding

affinity

and

interaction

patterns

of

major

essential

oil

constituents toward the Bcl-2 protein, while

cytotoxic

activity

was

evaluated

using

the

MTT assay on HeLa cervical cancer cells. The

findings

of

this

study

are

expected

to

provide

scientific

evidence

supporting

the

development

of

B. balsamifera

essential

oil

as a potential natural source of anti-cervical

cancer

agents

and

contribute

to

the

discovery

of

novel

apoptosis-targeting

therapeutics.

Materials and Methods

Materials, Apparatus, and Software

The

materials

used

in

this

study

included:

essential

oil

from

sembung

leaves

(

Blumea

balsamifera

),

cervical

cancer

cells

(HeLa),

DMEM

(Gibco,

2323000),

fetal

bovine

serum/FBS

(Gibco,

2445956RP),

antibiotic

and

antimycotic/ABAM

(Gibco,

2321098),

phosphate

buffer

saline

(PBS),

and

trypsin

(Gibco,

2391466).

The

equipment

used

included:

Biosafety

Cabinet

(Thermo

Scientific

1300

Series

A2

Class

II),

CO2

Incubator

(ARA

P170),

96

well

plate

(TPP,

92696),

Centrifuge

Tube

(SPL,

61015),

Haemocytometer,

Inverted

Microscope

(EVOS,

AMEX1000),

Plate

reader

(iMax

Biorad).

The

software

used

included:

GraphPad

Prism

10,

AutoDock

Vina

Tools-

1.5.7,

PyMOL

3.1.6.1,

and

BOVIA

Discovery

Studio 2022.

Molecular

docking

of

major

compounds

against Bcl-2

In

molecular

docking

analysis,

the

major

compounds Camphor, Caryophyllene, 7-epi-

Silphiperfol-5-ene,

and

Gamma-Eudesmol

(obtained from GC-MS analysis of sembung

leaf essential oil in the author's preliminary

study)

were

used

as

ligands.

Venetoclax

(ABT-199),

a

clinically

approved

Bcl-2

inhibitor,

was

used

as

the

positive

control

ligand.

The

Bcl-2

protein

with

PDB

codes

6QGK

was

used

as

the

receptor.

The

docking

process

was

performed

using

AutoDock

Vina

Tools,

PyMOL,

and

BIOVIA

Discovery

Studio

software.

The

three-

173

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

dimensional

structure

of

the

protein

was

obtained

from

the

Research

Collaboratory

for Structural Bioinformatics (RCSB) Protein

Data Bank (https://www.rcsb.org/), while the

ligand

structure

was

downloaded

from

PubChem(https://pubchem.ncbi.nlm.nih.go

v/).

Receptor

preparation

was

performed

with

PyMOL

by

removing

water

molecules

and

non-essential

residues

and

separating

the

native

ligand

from

the

receptor.

Next,

polar hydrogen atoms were added, and the

docking

process

was

performed

in

AutoDock Vina Tools. The interaction results

were

visualised

in

two

and

three

dimensions

using

BIOVIA

Discovery

Studio

.

The

docking

protocol

was

validated

by

redocking

the

native

ligand,

and

the

Root

Mean

Square

Deviation

(RMSD)

was

calculated.

The

anticancer

potential

of

essential

oil

from

sembung

leaves

against

cervical

cancer cells

Essential

oil

samples

were

dissolved

in

DMSO

to

obtain

a

stock

solution

of

1000

µg/mL, then diluted with DMEM medium to

obtain

test

concentrations

of

200,

100,

50,

25,

12.5

and

6.25

µg/mL,

with

the

final

DMSO concentration kept below 1% (v/v) to

avoid

cytotoxic

effects.

HeLa

cancer

cells

were grown in complete medium (DMEM +

10%

FBS

+

1%

antibiotic-antimycotic)

and

incubated

at

37

°C,

95%

humidity,

and

5%

CO

₂

until

they

reached

a

confluence

of

≥

80%.

Confluent

cells

were

harvested,

suspended,

and

seeded

in

a

96-well

plate

for 24 hours. After adding 20 µL of the test

solution,

the

culture

was

incubated

for

an

additional 48 hours. The medium was then

discarded,

and

the

cells

were

washed

with

100

µL

of

PBS.

This

was

followed

by

the

addition

of

MTT

reagent

(0.5

mg/mL)

and

incubation

for

4

hours.

The

formazan

crystals formed were dissolved with 100 µL

of

DMSO,

then

the

absorbance

was

measured

at

570

nm

using

a

microplate

reader. The

percentage of cell viability was

calculated

based

on

the

absorbance

value,

and

the

IC

₅₀

value

was

determined

using

GraphPad

Prism.

All

experiments

were

performed in quadruplicate, and the results

are

presented

as

mean

±

standard

deviation (SD). Untreated cells were used as

the growth control, while wells without cells

were

used

as

the

negative

control

(blank).

No

standard

anticancer

drug

was

included

as a positive control in this study.

Results and Discussions

Molecular

Docking

Analysis

against

BCL-2

Protein

Molecular

docking

testing

is

an

in

silico

method

used

to

predict

the

cytotoxic

activity

potential

of

the

major

compound

(Table

1)

in

sembung

leaves

essential

oil

against

cervical

cancer

cells.

The

receptor

selected was the Bcl-2 protein, because this

protein plays an

essential role in inhibiting

programmed

cell

death

(apoptosis).

Overexpression of the Bcl-2 protein results

in

a

decrease

in

the

ability

of

cells

to

undergo apoptosis, allowing abnormal cells

to

survive

and

potentially

accelerating

the

progression

of

precancerous

lesions

to

invasive

cervical

cancer.

In

addition,

Bcl-2

overexpression

has

been

reported

in

cervical

cancer

cells,

including

HeLa

cells,

where

it

contributes

to

apoptosis

resistance,

making

it

a

relevant

molecular

target for anticancer studies [8][9][10].

Table

1

.

Major

constituents

of

Blumea

balsamifera

leaf

essential

oil

identified

by

GC–MS.

No	Retention Time	Compound	Area (%)
1	14.766	Camphor	32.56
2	22.829	Caryophyllen	17.04
3	20.674	7-epi-Silphiperfol- 5-ene	5.33
4	28.246	Gamma-Eudesmol	5.01

174

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

The

Bcl-2

protein

structures

used

in

this

study

was

obtained

from

the

Protein

Data

Bank

(PDB

ID:

6QGK).

This

structure

was

selected

due

to

its

X-ray

diffraction

resolution of ≤2.5 Å, which indicates a high

level of accuracy in the atomic position and

active site conformation of the protein. This

value has a direct impact on the accuracy of

ligand-receptor

interaction

predictions,

so

that

the

docking

results

obtained

can

be

considered reliable for in silico analysis [11].

The

docking

protocol

was

validated

by

redocking

the

native

ligand

(J1Q)

into

the

Bcl-2

receptor

(PDB

ID:

6QGK),

resulting

in

an

RMSD

value

of

0.611

Å,

indicating

acceptable

docking

accuracy

because

the

RMSD value was below 2.0 Å. The grid box

parameters

for

the

Bcl-2

receptor

(PDB

ID:

6QGK)

were

defined

with

center

coordinates of x = 1.861, y = 0.439, and z =

19.153, and a grid size of 16 × 16 × 16 Å.

Table

2

presents

the

molecular

docking

results of the major compounds in essential

oils. The binding affinity values indicate the

strength of interaction between ligands and

target proteins, where more negative values

correspond

to

more

stable

ligand–protein

complexes

[12].

In

general,

binding

affinity

values

around

−5

kcal/mol

are

often

considered

to

indicate

relatively

weak

interactions, whereas values lower than

−7

kcal/mol may suggest stronger interactions.

These

classifications

are

based

on

previously

reported

literature

and

should

be interpreted cautiously, as binding affinity

alone

does

not

fully

represent

biological

activity [13,14].

Table

2

Molecular

docking

results

of

the

major compound in essential oils

Ligands	PDB ID	Binding Affinity (kcal/mol)
Camphor	6QGK	-5.745
Caryophyllene	6QGK	-7.077
7-epi-Silphiperfol-5- ene	6QGK	-7.311
Gamma-Eudesmol	6QGK	-7.083
Venetoclax	6QGK	-10.76

As

a

comparison,

Venetoclax,

a

clinically

approved Bcl-2 inhibitor used as a positive

control

ligand,

showed

a

binding

affinity

value

of

−10.76

kcal/mol,

indicating

a

very

strong

interaction

with

the

Bcl-2

protein.

Although

the

binding

affinity

values

of

the

major

compounds

from

sembung

leaf

essential

oil

were

lower

than

that

of

Venetoclax,

but

caryophyllene,

7-epi-

silphiperfol-5-ene,

and

γ-eudesmol

still

showed

relatively

strong

interactions,

with

binding

affinity

values

of

−7.077,

−7.311,

and

−7.083

kcal/mol,

respectively.

Meanwhile,

camphor

exhibited

a

binding

affinity value of -5.745 kcal/mol, which may

be classified as a moderate interaction.

Figure 1.

Interaction of camphor ligand with 6QGK receptor (A) 2D Interaction Diagram (B) 3D

ligand-receptor complex (C) View of binding pocket surface

175

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

Figure 1 shows the results of the interaction

between

the

caryophyllene

ligand

and

the

Bcl-2

receptor

(PDB

ID:

6QGK).

The

2D

interaction diagram shows alkyl and π–alkyl

interactions

(light

purple)

with

residues

Met115,

Ala149,

Phe104,

and

Phe112.

In

addition, van der Waals interactions (green)

were

observed

with

residues

Asp111,

Tyr108, Phe153, and Leu137. The 3D ligand–

receptor

complex

shows

that

the

ligand

is

positioned

within

the

binding

pocket

surrounded by α-helical structures (red) and

loop

regions

(green

and

gray).

The

surface

view

indicates

that

the

ligand

occupies

a

hydrophobic

binding

pocket,

which

may

contribute

to

the

stability

of

the

caryophyllene–6QGK

complex

and

support

the

binding

affinity

obtained

from

the

docking results.

Figure

2.

Interaction of the caryophyllene ligand with the 6QGK receptor (A) 2D interaction

diagram (B) 3D ligand-receptor complex (C) View of the binding pocket surface

Figure

2

shows

the

interaction

of

the

caryophyllene

compound

with

the

Bcl-2

receptor

(PDB

ID:

6QGK).

The

ligand

interacts

via

π–alkyl

forces

(light

purple)

with

residues

Phe104,

Tyr108,

Phe112,

Met115, Val133, and Leu137, as well as van

der

Waals

forces

(green)

with

residues

Asp111

and

Ala149.

The

ligand

appears

to

be

stably

oriented

within

the

hydrophobic

pocket,

which

plays

a

crucial

role

in

maintaining

the

caryophyllene-6QGK

complex and enhancing the binding affinity,

as indicated by the docking results.

Figure

3.

Interaction

of

the

7-epi-Silphiperfol-5-ene

ligand

with

the

6QGK

receptor

(A)

2D

Interaction Diagram (B) 3D ligand-receptor complex (C) View of the binding pocket surface

176

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

Figure

4.

Interaction

of

the

Gamma-Eudesmol

ligand

with

the

6QGK

receptor

(A)

2D

Interaction Diagram (B) 3D Ligand-Receptor Complex (C) View of the binding pocket surface

Figure 3 shows the interaction of the 7-epi-

Silphiperfol-5-ene

ligand

compound

with

the

Bcl-2

receptor

(PDB

ID:

6QGK)

through

van der Waals bonds (green) with residues

Asp111

and

Phe150,

as

well

as

π–sigma

interactions (dark purple), alkyl, and π–alkyl

(light

purple)

interactions

with

residues

Met115,

Leu137,

Ala149,

Val133,

Phe112,

Phe104,

Phe153,

and

Tyr108.

These

interactions suggest that the stability of the

7-epi-Silphiperfol-5-ene–hydrophobic

interactions

primarily

support

the

6QGK

complex,

while

the

other

residues

contribute

through

weak

nonpolar

interactions.

Figure

4

shows

the

interaction

of

the

γ-

eudesmol

compound

with

the

Bcl-2

receptor

(PDB

ID:

6QGK).

The

ligand

interacts

via

π–sigma

forces

(dark

purple)

with

residue

Phe112,

as

well

as

π–alkyl

forces

(light

purple)

with

Phe104,

Tyr108,

Met115,

Val133,

Leu137,

Ala149,

and

Phe153.

In

addition,

there

is

a

van

der

Waals interaction (green) with Asp111. This

combination

of

hydrophobic

interactions

plays

an

important

role

in

maintaining

the

stability and binding affinity of the complex.

Based

on

the

results

of

molecular

docking

tests

on

the

major

compound

of

sembung

leaves,

which

were

reviewed

in

terms

of

RMSD

values,

binding

affinity

energy,

and

the

types

of

interactions

formed

between

ligands and amino acid residues such as π–

sigma,

π–alkyl,

and

van

der

Waals,

it

was

found

that

sembung

plants

may

have

potential

as

anticancer

agents,

particularly

for

cervical

cancer.

Therefore,

further

in

vitro research using the MTT assay method

is required to validate these findings.

The

anticancer

potential

of

essential

oil

from

Sembung

Leaves

against

Cervical

Cancer Cells

Anticancer testing was conducted using the

MTT

(Microculture

Tetrazolium

Test)

method, which aims to determine the ability

of

essential

oils

to

inhibit

cell

growth.

This

test is based on cell viability percentage and

the determination of the IC

50

value. The cell

viability

percentage

at

a

given

sample

absorbance value determines the IC

50

value.

The sample absorbance values from various

concentrations can be used to calculate the

cell

viability

percentage,

which

represents

the

number

of

cells

that

survive

after

exposure to a test sample. The test results

are shown in Figure 5.

Figure 5a shows that the absorbance value

decreases

with

increasing

concentration.

This

indicates

that

higher

concentrations

177

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

contain

greater

amounts

of

active

compounds

in

the

sample,

which

may

inhibit

cell

growth.

A

similar

trend

is

observed

in

Figure

5b,

where

cell

viability

decreases

as

the

concentration

increases.

The

decrease

in

absorbance

and

cell

viability

is

associated

with

reduced

activity

of

succinate

dehydrogenase,

which

catalyses

the

reduction

of

tetrazolium

salt

to

formazan

crystals,

as

cell

viability

decreases at higher concentrations.

Figure

5.

MTT assay results of

Blumea

balsamifera

leaf essential oil on HeLa cervical cancer

cells.

(a)

Absorbance

at

570

nm

after

treatment

with

different

concentrations.

(b)

Dose–

response

curve

showing

cell

viability (%)

versus

log

concentration,

yielding an

IC

₅₀

value

of

32.90 μg/mL.

Figure

5b

shows

that

at

the

highest

concentration (200 µg/mL), cell viability was

16.86%, indicating that cell growth was not

completely

inhibited

at

this

concentration.

Meanwhile,

at

the

lowest

concentration

(6.25

µg/mL),

the

cell

viability

percentage

was 88.25%, indicating that cell growth was

inhibited

by

approximately

12%

at

this

concentration. The test results showed that

the essential oil exhibited cytotoxic activity,

with

an

IC

₅₀

value

of

32.90

µg/mL,

which

may

be

classified

as

moderate

cytotoxic

activity

based

on

the

National

Cancer

Institute

(NCI)

criteria,

where

IC

₅₀

values

between

20–100

µg/mL

are

considered

moderate [15][16]. This IC

₅₀

value indicates

that the essential oil has potential cytotoxic

activity against HeLa cervical cancer cells.

However,

the

absence

of

a

standard

anticancer

drug

(e.g.,

doxorubicin)

as

a

positive

control

represents

a

limitation

of

this

study

and

should

be

considered

in

interpreting

the

cytotoxicity

results.

The

molecular

docking

results

suggest

that

the

major

compounds

of

sembung

leaf

178

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

essential

oil

may

interact

with

the

Bcl-2

protein. However, the relationship between

Bcl-2

binding

affinity

and

the

observed

cytotoxic

activity

in

the

MTT

assay

has

not

been experimentally confirmed in this study

and

requires

further

investigation.

Nevertheless,

the

observed

cytotoxic

activity,

together

with

the

molecular

docking results, suggests that the essential

oil

has

potential

biological

activity

against

the

Bcl-2

target.

Further

studies

using

standard anticancer agents and mechanistic

analysis

are

recommended

to

strengthen

these findings.

This

cytotoxic

ability

is

attributed

to

the

active

compounds

in

sembung

leaves

essential

oil,

which

are

primarily

terpenoid

compounds.

Terpenoid

compounds

have

been

widely

reported

to

exhibit

cytotoxic

and

pharmacological

bioactivity,

such

as

anticancer

[17][18].

Additionally,

compounds

in

essential

oils

have

been

reported

to

inhibit

cancer

cell

growth

by

inducing apoptosis and increasing oxidative

stress, as reported in previous studies. This

mechanism

is

suggested

to

contribute

to

cancer

cell

damage

[19][20][21].

However,

these proposed mechanisms have not been

experimentally confirmed in this study and

are presented based on previously reported

findings,

while

further

studies

are

needed

to validate these mechanisms.

Conclusions

The

results

of

this

study

suggest

that

sembung

leaf

essential

oil

has

potential

anticancer

activity

against

HeLa

cervical

cancer

cells

based

on

molecular

docking

and

MTT

assay

results.

Caryophyllene,

7-

epi-silphiperfol-5-ene,

and

γ-eudesmol

showed

relatively

strong

interactions

with

the

Bcl-2

protein,

while

the

essential

oil

exhibited

moderate

cytotoxic

activity

with

an

IC

₅₀

value

of

32.90

µg/mL.

However,

further

studies

are

needed

to

confirm

the

proposed anticancer mechanism.

Acknowledgement

This

study

was

conducted

as

a

part

of

the

research

activities

supported

by

the

Directorate

of

Research,

Technology,

and

Community

Service

(DRTPM),

Ministry

of

Education,

Culture,

Research,

and

Technology

of

the

Republic

of

Indonesia,

under

the

Penelitian

Dosen

Pemula

(2025)

scheme. The authors also acknowledge the

support

and

facilitation

provided

by

the

Institute

for

Research

and

Community

Service

(LPPM),

Universitas

Teknologi

Nusantara.

Author Contributions

Conceptualization, Rahmi Vika Ulia and Toto

Raharto;

Methodology,

Rahmi

Vika

Ulia;

Software, Toto Raharto and Yoan De Nanda

Herru;

Validation,

Rahmi

Vika

Ulia,

Suryati,

and

Adlis

Santoni;

Formal

Analysis,

Toto

Raharto; Investigation, Rahmi Vika Ulia, Toto

Raharto, Suryati, Adlis Santoni, and Yoan De

Nanda

Herru;

Resources,

Rahmi

Vika

Ulia

and

Toto

Raharto;

Data

Curation,

Yoan

De

Nanda

Herru;

Writing

–

Original

Draft

Preparation,

Rahmi

Vika

Ulia;

Writing

–

Review & Editing, Suryati; Visualization, Adlis

Santoni;

Supervision,

Rahmi

Vika

Ulia;

Project

Administration,

Toto

Raharto;

Funding Acquisition, Rahmi Vika Ulia.

Conflict of Interest

The authors declare no conflict of interest.

References

[1].

Widhiantara,

G.,

and

Jawi,

I.M.

Phytochemical

composition

and

health

properties

of

Sembung

plant

(

Blumea

balsamifera

):

A

review.

Veterinary

World.

2021,

14(5),

1185–1196.

DOI:

10.14202/vetworld.2021.1185-1196.
[2].

Norikura, T., Kojima-Yuasa, A., Shimizu, M.,

Huang, X., Xu, S., Kametani, S., Rho, S.-N.,

179

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

Kennedy,

D.

O.,

and

Matsui-Yuasa,

I.

Anticancer

activities

and

mechanisms

of

Blumea

balsamifera

extract

in

hepatocellular

carcinoma

cells.

The

American Journal of Chinese Medicine.

2008,

36(2),

411-424).

DOI:

10.1142/S0192415X08005862

.
[3].

Ismail, N. A., Matawali, A., Lee, P.-C., How,

S.-E., Lee, B. H., Goh, L. P. W., and Gansau,

J. A.

Blumea balsamifera

(L.) DC. Elicit Anti-

Kinase,

Anti-Phosphatase

and

Cytotoxic

Activities

against

Acute

Promyelocytic

Leukemia Cells (HL-60).

Tropical Journal of

Natural

Product

Research

.

2023,

5(4),

656-

660. DOI:

10.26538/tjnpr/v5i5.8

.
[4].

Nurlailiyah,

F.

A.,

Dwijayanti,

D.

R.,

Hermanto, F. E., Masruri, M., and Widodo,

N.

Cytotoxicity

of

Blumea

balsamifera

on

A549

Lung

Cancer

Cells

 

:

Integrating

in

Vitro Analysis with Computational Study of

AKT-1 Inhibition.

Tropical Journal of Natural

Product Research

. 2025, 9(2), 495–503. DOI:

10.26538/tjnpr/v9i2.12

.
[5].

World Health Organization. Global Cancer

Observatory. 2022.
[6].

Permatasari,

H.

K.,

Wewengkang,

D.

S.,

Tertiana,

N.

I.,

Muslim,

F.

Z.,

Yusuf,

M.,

Baliulina, S. O., Daud, V. P. A., Setiawan, A.

A., and Nurkolis, F. Anti-cancer properties

of

Caulerpa

racemosa

by

altering

expression of Bcl-2, BAX, cleaved caspase

3

and

apoptosis

in

HeLa

cancer

cell

culture.

Frontiers

in

Oncology

.

2022,

12,

964816. DOI:

10.3389/fonc.2022.964816

.
[7].

Leisching,

G.,

Loos,

B.,

Botha,

M.,

and

Engelbrecht, A.-M. Bcl-2 confers survival in

cisplatin

treated

cervical

cancer

cells:

Circumventing

cisplatin

dose-dependent

toxicity

and

resistance.

Journal

of

Translational

Medicine

.

2015,

13(1),

328.

DOI:

10.1186/s12967-015-0689-4
[8].

Yumol, J., Gabrielli, B., Tayyar, Y., McMillan,

N.

A.,

and

Idris,

A.

Smart

drug

combinations

for

cervical

cancer:

Dual

targeting

of

Bcl-2

family

of

proteins

and

aurora kinases.

American Journal of Cancer

Research

.

2020,

10(10),

3406–3414.

PMID:

33163279
[9].

Abdul Rahman, S. F., Muniandy, K., Soo, Y.

K.,

Hamdan,

M.

A.,

Shunmugam,

L.,

and

Idris, A. Co-inhibition of BCL-XL and MCL-1
with

selective

BCL-2

family

inhibitors

enhances

cytotoxicity

of

cervical

cancer

cell

lines.

Biochemistry

and

Biophysics

Reports

.

2020,

22,

100756.

DOI:

10.1016/j.bbrep.2020.100756

.
[10].

Tuohetimulati,

G.,

Zhu,

M.,

Chen,

J.,

and

Niyazi,

M.

Expressions

and

clinical

significance

of

Bcl-2,

Bcl-xL

and

c-IAP1

protein

in

cervical

cancer.

International

Journal

of

Clinical

and

Experimental

Medicine

. 2018, 11(11), 12361–12367.
[11].

Ferreira de Freitas, R., and Schapira, M. A

systematic

analysis

of

atomic

protein–

ligand

interactions

in

the

PDB.

Medicinal

Chemistry

Communications

.

2017,

8(10),

1970–1981. DOI:

10.1039/C7MD00381A

.
[12].

Ramírez, D., and Caballero, J. Is It Reliable

to

Take

the

Molecular

Docking

Top

Scoring

Position

as

the

Best

Solution

without

Considering

Available

Structural

Data?.

Molecules.

2018,

23(5),

1038.

DOI:

10.3390/molecules23051038

.
[13].

Sharma, A., Vora, J., Patel, D., Sinha, S., Jha,

P. C., and Shrivastava, N. Identification of

natural inhibitors against prime targets of

SARS-CoV-2

using

molecular

docking,

molecular

dynamics

simulation

and

MM-

PBSA

approaches.

Journal

of

Biomolecular

Structure and Dynamics

. 2022, 40(7), 3296–

3311.

DOI:

10.1080/07391102.2020.1846624

.
[14].

Ivanova,

L.,

and

Karelson,

M.

The

Impact

of Software Used and the Type of Target

Protein

on

Molecular

Docking

Accuracy.

Molecules

.

2022,

27(24),

9041.

DOI:

10.3390/molecules27249041

.
[15].

Niksic,

H.,

Becic,

F.,

Koric,

E.,

Gusic,

I.,

Omeragic, E., Muratovic, S., Miladinovic, B.,

and

Duric,

K.

Cytotoxicity

screening

of

Thymus

vulgaris

L.

essential

oil

in

brine

shrimp

nauplii

and

cancer

cell

lines.

Scientific Reports

.

2021,

11(1),

13178.

DOI:

10.1038/s41598-021-92679-x

.
[16].

Zulkipli,

N.

N.,

Rahman,

S.

A.,

Taib,

W.

R.

W., Razali, R. M., Ismail, I., Ahmad, W. A. N.

W., and Daud, C. K. D. C. The cytotoxicity

effect

and

identification

of

bioactive

compounds of

Prismatomeris glabra

crude

leaf

extracts

against

breast

cancer

cells.

Beni-Suef

University

Journal

of

Basic

and

Applied

Sciences

.

2024,

13(1),

33.

DOI:

180

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

10.1186/s43088-024-00490-0.
[17].

Kamran, S., Sinniah, A., Abdulghani, M. A.

M.,

and

Alshawsh,

M.

A.

Therapeutic

Potential

of

Certain

Terpenoids

as

Anticancer

Agents:

A

Scoping

Review.

Cancers

.

2022,

14(5),

1100.

DOI:

10.3390/cancers14051100

.
[18].

Wu,

Z.

L.,

Li,

N.,

Huang,

J.

L.,

Sang,

H.

Y.,

and

Chen,

X.

Essential

oil

and

its

major

compounds

from

oil

camphor

inhibit

human lung and breast cancer cell growth

by cell-cycle arresting.

International Journal

of Clinical and Experimental Medicine

. 2016,

9(7), 12852–12861.
[19].

Madrid, A., Silva, V., Russo, A., Moller, A. C.,

Sánchez,

E.,

Villena,

J.,

Jara-Gutiérrez,

C.,

and

Montenegro,

I.

Selective

cytotoxicity

of

Gomortega

keule

essential

oil

through

a

ROS-mediated

pro-apoptotic

mechanism.

Frontiers

in

Pharmacology

.
2025,

16,

1722619.

DOI:

10.3389/fphar.2025.1722619

.
[20].

Abdoul-Latif, F. M., Ainane, A., Aboubaker,

I.

H.,

Mohamed,

J.,

and

Ainane,

T.

Exploring the Potent Anticancer Activity of

Essential

Oils

and

Their

Bioactive

Compounds:

Mechanisms

and

Prospects

for

Future

Cancer

Therapy.

Pharmaceuticals

.

2023,

16(8),

1086.

DOI:

10.3390/ph16081086.
[21].

Bayala,

B.,

Bassole,

I.

H.

N.,

Scifo,

R.,

Gnoula,

C.,

Morel,

L.,

Lobaccaro,

J.

M.

A.,

Simpore,

J.,

and

Fancellu,

A.

Anticancer

activity of essential oils and their chemical

components

–

a

review.

American

Journal

of

Cancer

Research

.

2014,

4(6),

591–607.

PMID: 25520854.

181

RV Ulia et al.,

Chempublish Journal, 10(1) 2026, 171-181

In silico and In vitro Study of The Anticancer Potential of Essential Oil from Sembung ( Blumea balsamifera ) Leaves against HeLa Cervical Cancer Cells

Rahmi Vika Ulia 1 , Toto Raharto 1 , Suryati 2 , Adlis Santoni 2 , Yoan De Nanda Herru 3

A bstract

Graphical Abstract

Introduction

In silico and In vitro Study of The Anticancer Potential of Essential

Oil from Sembung (

Blumea

balsamifera

) Leaves against HeLa Cervical

Cancer Cells

Rahmi Vika Ulia

1

, Toto Raharto

1

, Suryati

2

,

Adlis Santoni

2

,

Yoan De Nanda Herru

3

A

bstract