Uji Aktivitas Antikanker Ekstrak Etanol Buah Andaliman (Zanthoxylum acanthopodium DC.) Terhadap Sel Kanker Serviks
UJI AKTIVITAS ANTIKANKER EKSTRAK ETANOL
BUAH ANDALIMAN (Zanthoxylum acanthopodium DC.)
TERHADAP SEL KANKER SERVIKS
Abstrak
Kanker serviks merupakan salah satu jenis kanker yang menjadi penyebab
kematian terbesar di Indonesia pada wanita. Penyebab utama kanker serviks
adalah Human Papilloma Virus (HPV). Andaliman (Zanthoxylum acanthopodium
DC.) adalah salah satu tanaman yangdikenal dalam masyarakat Batak, tergolong
dalam tanaman liar yang belum banyak dimanfaatkan sebagai tanaman obat. Buah
andaliman diyakini memiliki potensi antikanker. Tujuan penelitian ini adalah
untuk mengetahui karakteristik simplisia buah andaliman, menghitung nilai IC50
ekstrak terhadap sel HeLa dan menghitung indeks selektivitas ekstrak etanol buah
andaliman.
Ekstrak diperoleh dengan maserasi menggunakan pelarut etanol 96%. Uji
aktivitas sitotoksik EEBA secara in vitro terhadap sel HeLa dan sel Vero
menggunakan metode MTT. Sel HeLa ditumbuhkan dalam media kultur RPMI
pada plate 96-sumuran kemudian diberi ekstrak etanol buah andaliman dengan
seri konsentrasi 500 µg/mL; 250 µg/mL; 125 µg/mL; 62,5 µg/mL; 31,25 µg/mL
dan 15,625 µg/mL.Sel Vero ditumbuhkan dalam media kultur M199 pada96sumuran kemudian diberi ekstrak etanol buah andaliman dengan seri konsentrasi
1000 µg/mL; 500 µg/mL; 250 µg/mL; 125 µg/mL; 62,5 µg/mL dan 31,25 µg/mL.
Hasil pengujian dibaca dengan ELISA reader pada 595 nm dan dianalisis dengan
SPSS 19 kemudian dihitung indeks selektivitasnya.
Hasilskrining fitokimia terhadap simplisia dan ekstrak etanol buah
andaliman diperoleh senyawa kimia golongan alkaloid, flavonoid, tanin,
glikosida, dan steroid/triterpenoid. Hasil pemeriksaan karakteristik simplisia
diperoleh kadar air 6,30%; kadar sari larut air 12,41%; kadar sari larut dalam
etanol 16,28%; kadar abu total 4,62% dan kadar abu tidak larut asam 0,20%. Hasil
pengujian aktivitas sitotoksik ekstrak etanol buah andaliman terhadap sel HeLa
menunjukkan nilai IC50sebesar 42,462 µg/mL dan pada sel Vero sebesar
258,167µg/mL. Hal ini menunjukkan bahwa ekstrak etanol buah andaliman
memiliki efek sitotoksisitas yang poten terhadap sel kanker serviks HeLa. EEBA
memiliki indeks selektivitas sebesar 6,08. Hal ini berarti EEBA bekerja selektif
yaitu hanya menghambat sel kanker serviks tanpa mematikan sel normal (Vero).
Kata Kunci: MTT, sitotoksisitas, Sel HeLa, Sel Vero, Zanthoxylum
acanthopodium DC.
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ANTICANCER ACTIVITY OF ETHANOL EXTRACT FROM
ANDALIMAN FRUIT (Zanthoxylum acanthopodium DC.)
ON CERVICAL CANCER CELLS
Abstract
Cervical cancer is one type of the largest cancer that becomes largest cause
of death woman in Indonesia.The main cause of cervical cancer is Human
Papilloma Virus (HPV).Andaliman ( Zanthoxylum acanthopodium DC.) is one of
the plant which well known within the community Bataknese were classified as a
part wild plants that have not been widely used as a medicinal plant. Fruit
andaliman does believed have anticancer potential. The purpose of this research
were to known characteristic of andaliman fruit simplex, to determinated the IC50
value of extract on HeLa cell and determinated the selectivity index of EEBA.
Extraction were used maceration method with 96% ethanol. Cytotoxic
effect was tested in vitro on HeLa and Vero cells line using MTT assay. HeLa
cells were seeded in RPMI medium at 96-well plate and then were given
andaliman fruitethanolic extract with concentration 500 µg/mL; 250 µg/mL; 125
µg/mL; 62.5 µg/mL;31.25 µg/mL and 15.625 µg/mL. While Vero cells were
seeded in M199 culture medium at 96-well plate and then were given extract with
concentration 1000 µg/mL; 500 µg/mL; 250 µg/mL; 125 µg/mL; 62.5µg/mL and
31.25 µg/mL The absorbanceswere read by ELISA reader at ƛ 595 nm and
analysed with SPSS 19 and calculated selectivity index.
The results of photochemical screening from simplex and ethanol extract
of Andaliman fruit contains alkaloid, flavonoids, glikosids, tannins and
steroids/triterpenoids. The result of the simplex characteristics were showed that
6.30% water content; 12.41% water-soluble extract content; 16.28% ethanolsoluble extract content; 4.62% total ash and 0.20% acidic unsoluble ash. The
cytotoxic activity assay was showed that ethanol extract of Andaliman fruit on
HeLa cells have IC50 values of 42.462 µg/mL and for Vero cells was showed IC50
values of 258.167 µg/mL. Andaliman fruits ethanolic extract was exhibited
cytotoxic effect against cervical cancer HeLa cells. EEBA have selectivity index
of 6.08. Ethanolic extract of andaliman fruit have selective activity cervical cancer
cell without inhibit the normal cell (Vero).
Keywords: MTT, Cytotoxicity, HeLa Cells, Vero Cells, Zanthoxylum
acanthopodium DC.
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BUAH ANDALIMAN (Zanthoxylum acanthopodium DC.)
TERHADAP SEL KANKER SERVIKS
Abstrak
Kanker serviks merupakan salah satu jenis kanker yang menjadi penyebab
kematian terbesar di Indonesia pada wanita. Penyebab utama kanker serviks
adalah Human Papilloma Virus (HPV). Andaliman (Zanthoxylum acanthopodium
DC.) adalah salah satu tanaman yangdikenal dalam masyarakat Batak, tergolong
dalam tanaman liar yang belum banyak dimanfaatkan sebagai tanaman obat. Buah
andaliman diyakini memiliki potensi antikanker. Tujuan penelitian ini adalah
untuk mengetahui karakteristik simplisia buah andaliman, menghitung nilai IC50
ekstrak terhadap sel HeLa dan menghitung indeks selektivitas ekstrak etanol buah
andaliman.
Ekstrak diperoleh dengan maserasi menggunakan pelarut etanol 96%. Uji
aktivitas sitotoksik EEBA secara in vitro terhadap sel HeLa dan sel Vero
menggunakan metode MTT. Sel HeLa ditumbuhkan dalam media kultur RPMI
pada plate 96-sumuran kemudian diberi ekstrak etanol buah andaliman dengan
seri konsentrasi 500 µg/mL; 250 µg/mL; 125 µg/mL; 62,5 µg/mL; 31,25 µg/mL
dan 15,625 µg/mL.Sel Vero ditumbuhkan dalam media kultur M199 pada96sumuran kemudian diberi ekstrak etanol buah andaliman dengan seri konsentrasi
1000 µg/mL; 500 µg/mL; 250 µg/mL; 125 µg/mL; 62,5 µg/mL dan 31,25 µg/mL.
Hasil pengujian dibaca dengan ELISA reader pada 595 nm dan dianalisis dengan
SPSS 19 kemudian dihitung indeks selektivitasnya.
Hasilskrining fitokimia terhadap simplisia dan ekstrak etanol buah
andaliman diperoleh senyawa kimia golongan alkaloid, flavonoid, tanin,
glikosida, dan steroid/triterpenoid. Hasil pemeriksaan karakteristik simplisia
diperoleh kadar air 6,30%; kadar sari larut air 12,41%; kadar sari larut dalam
etanol 16,28%; kadar abu total 4,62% dan kadar abu tidak larut asam 0,20%. Hasil
pengujian aktivitas sitotoksik ekstrak etanol buah andaliman terhadap sel HeLa
menunjukkan nilai IC50sebesar 42,462 µg/mL dan pada sel Vero sebesar
258,167µg/mL. Hal ini menunjukkan bahwa ekstrak etanol buah andaliman
memiliki efek sitotoksisitas yang poten terhadap sel kanker serviks HeLa. EEBA
memiliki indeks selektivitas sebesar 6,08. Hal ini berarti EEBA bekerja selektif
yaitu hanya menghambat sel kanker serviks tanpa mematikan sel normal (Vero).
Kata Kunci: MTT, sitotoksisitas, Sel HeLa, Sel Vero, Zanthoxylum
acanthopodium DC.
vi
ANTICANCER ACTIVITY OF ETHANOL EXTRACT FROM
ANDALIMAN FRUIT (Zanthoxylum acanthopodium DC.)
ON CERVICAL CANCER CELLS
Abstract
Cervical cancer is one type of the largest cancer that becomes largest cause
of death woman in Indonesia.The main cause of cervical cancer is Human
Papilloma Virus (HPV).Andaliman ( Zanthoxylum acanthopodium DC.) is one of
the plant which well known within the community Bataknese were classified as a
part wild plants that have not been widely used as a medicinal plant. Fruit
andaliman does believed have anticancer potential. The purpose of this research
were to known characteristic of andaliman fruit simplex, to determinated the IC50
value of extract on HeLa cell and determinated the selectivity index of EEBA.
Extraction were used maceration method with 96% ethanol. Cytotoxic
effect was tested in vitro on HeLa and Vero cells line using MTT assay. HeLa
cells were seeded in RPMI medium at 96-well plate and then were given
andaliman fruitethanolic extract with concentration 500 µg/mL; 250 µg/mL; 125
µg/mL; 62.5 µg/mL;31.25 µg/mL and 15.625 µg/mL. While Vero cells were
seeded in M199 culture medium at 96-well plate and then were given extract with
concentration 1000 µg/mL; 500 µg/mL; 250 µg/mL; 125 µg/mL; 62.5µg/mL and
31.25 µg/mL The absorbanceswere read by ELISA reader at ƛ 595 nm and
analysed with SPSS 19 and calculated selectivity index.
The results of photochemical screening from simplex and ethanol extract
of Andaliman fruit contains alkaloid, flavonoids, glikosids, tannins and
steroids/triterpenoids. The result of the simplex characteristics were showed that
6.30% water content; 12.41% water-soluble extract content; 16.28% ethanolsoluble extract content; 4.62% total ash and 0.20% acidic unsoluble ash. The
cytotoxic activity assay was showed that ethanol extract of Andaliman fruit on
HeLa cells have IC50 values of 42.462 µg/mL and for Vero cells was showed IC50
values of 258.167 µg/mL. Andaliman fruits ethanolic extract was exhibited
cytotoxic effect against cervical cancer HeLa cells. EEBA have selectivity index
of 6.08. Ethanolic extract of andaliman fruit have selective activity cervical cancer
cell without inhibit the normal cell (Vero).
Keywords: MTT, Cytotoxicity, HeLa Cells, Vero Cells, Zanthoxylum
acanthopodium DC.
vii