SKRIPSI
TUTUT CHYNTIA RIZCA
UJI SITOTOKSISITAS FRAKSI ETANOL AKAR
Jatropha gossypifolia L. TERHADAP SEL
KANKER PAYUDARA MCF-7 DENGAN
METODE MTT ASSAY
PROGRAM STUDI FARMASI
FAKULTAS ILMU KESEHATAN
UNIVERSITAS MUHAMMADIYAH MALANG
2017
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KATA PENGANTAR
Assalamu’alaikum warahmatullahi wabarakatuh
Alhamdulillahirobbil’alamin, atas segala nikmat iman, islam, kesempatan, serta
kekuatan yang telah diberikan Allah Subhanahuwata’ala sehingga penulis dapat
menyelesaikan skripsiyang berjudul “UJI SITOTOKSISITAS FRAKSI
ETANOL AKAR Jatropha gossypifolia L. TERHADAP SEL KANKER
PAYUDARA MCF-7 DENGAN METODE MTT ASSAY”. Shalawat beriring
salam untuk tuntunan dan suri tauladan Rasulullah Shallallahu’alaihiwasallam
beserta keluarga dan sahabat beliau yang senantiasa menjunjung tinggi nilai-nilai
islam yang sampai saat ini dapat dinikmati oleh seluruh manusia di penjuru dunia.
Skripsi ini disusun untuk memenuhi salah satu syarat untuk mencapai gelar
sarjana Farmasi pada Program Studi Farmasi Fakultas Ilmu Kesehatan Universitas
Muhammadiyah Malang. Selama kegiatan penulisan skripsi, penulis tidak luput
dari kendala yang dihadapi. Akan tetapi, kendala-kendala tersebut dapat diatasi
penulis berkat adanya bantuan, bimbingan dari berbagai pihak. Oleh karena itu
penulis ingin menyampaikan rasa terima kasih dan penghargaan yang sebesar-
besarnya kepada:
1. Siti Rofida, S.Si., M.Farm., Apt selaku dosen pembimbing I dan Ahmad
Shobrun Jamil, S.Si., MP. selaku dosen pembimbing II atas saran,
bimbingan, arahan serta saran dan motivasi kepada penulis sampai
terselesaikannya skripsi ini.
2. Engrid Juni Astuti,S.Farm., M.Farm., Apt selaku dosen penguji I dan
Andri Tilaqza, S.Farm., M.Farm., Apt. selaku selaku dosen penguji II yang
telah banyak memberikan saran dan masukan demi kesempurnaan skripsi
ini.
3. Yoyok Bekti Prasetyo, S.Kep, M.Kep., Sp.Kom selaku Dekan Fakultas
Ilmu Kesehatan Universitas Muhammadiyah Malang.
4. Nailis Syifa, S.Farm., M.Sc., Apt., selaku Ketua Program Studi Farmasi
Universitas Muhammadiyah Malang.
5. Ika Ratna Handayanti S.Farm., M.Sc., Apt selaku dosen wali yang telah
memberikan saran, nasehat, motivasi, dan bimbingannya selama ini.
6. Laboratorium Parasitologi Fakultas Kedokteran Universitas Gajah Mada
khususnya Rumbiwati, ST., M.Sc. dan Prof. dr. Supargiyono, DTM&H.,
v
SU., Ph.D., SpParK. yang membantu dan mengarahkan selama proses
penelitian.
7. Sovia Aprina Basuki S.Farm., M.Si., Apt. selaku Kepala Laboratorium
Farmasi yang telah memberikan fasilitas selama melakukan penelitian.
8. Keluargaku tercinta Ayah, Ibu, dan Adik, yang tidak pernah lelah
memberikan motivasi, nasehat, dan kasih sayangnya tiada henti yang telah
mendo’akan dan mendampingi sampai dengan selesai mengerjakan
skripsi.
9. Dosen-dosen Farmasi Universitas Muhammadiyah Malang dengan
dedikasinya dalam dunia pendidikan atas ilmu selama menempuh kuliah.
10. Untuk semua anggota tata usaha program studi farmasi fakultas ilmu
kesehatan universitas muhammadiyah malang, yang telah banyak
membantu untuk kebutuhan administrasi kelengkapan skripsi.
11. Teman terbaikku (Deviana, Yeni, Parista, teman-teman Farmasi C),
Teman penelitian seperjuangan (Wulan, Linda, Fika, Irul, Nikma, Risa,
Tria, dan Fatma) yang selalu memberikan motivasi, bantuan, serta nasehat.
Terimakasih sudah menemani selama ini, kebersamaan ini tidak akan
pernah terlupakan.
12. Untuk semua pihak yang belum disebutkan namanya, penulis mohon maaf
dan terima kasih yang sebesar-besarnya. Semua keberhasilan ini tak luput
dari bantuan dan doa yang telah kalian semua berikan.
Tentunya skripsi ini masih jauh dari kesempurnaan. Karena pada dasarnya Penulis
adalah manusia biasa yang memiliki keterbatasan serta kemampuan yang berbeda-beda.
Oleh karena itu, penulis sangat berharap akan adanya masukan berupa saran dan kritik
yang sifatnya membangun untuk perbaikan di masa mendatang, Semoga Allah senantiasa
melimpahkan karunia-Nya dan semoga skripsi ini bermanfaat bagi masyarakat. Aamiin.
Wassalamu’alaikum warohmatullohi wabarokatuh
Malang, 20 Juni 2017
Penyusun,
Tutut Chyntia Rizca
x
DAFTAR ISI
Halaman
LEMBAR PENGESAHAN .................................................................................. ii
LEMBAR PENGUJIAN ...................................................................................... iii
KATA PENGANTAR .......................................................................................... iv
RINGKASAN ....................................................................................................... vi
ABSTRAK .......................................................................................................... viii
ABSTRACT .......................................................................................................... ix
DAFTAR ISI .......................................................................................................... x
DAFTAR TABEL .............................................................................................. xiv
DAFTAR GAMBAR ........................................................................................... xv
DAFTAR LAMPIRAN ...................................................................................... xvi
DAFTAR SINGKATAN ................................................................................... xvii
BAB I PENDAHULUAN ...................................................................................... 1
1.1 Latar Belakang .................................................................................................. 1
1.2 Rumusan Masalah ............................................................................................. 4
1.3 Tujuan ............................................................................................................... 4
1.4 Manfaat Penelitian ........................................................................................... 4
1.4.1 Segi Akademik ........................................................................................ 4
1.4.2 Segi Masyarakat ...................................................................................... 4
BAB II TINJAUAN PUSTAKA ........................................................................... 6
2.1 Tinjauan Tanaman ............................................................................................. 6
2.1.1 Klasifikasi Tanaman ............................................................................... 6
2.1.2 Sinonim ................................................................................................... 6
2.1.3 Morfologi Tanaman ................................................................................ 7
xi
2.1.4 Daerah Asal dan Penyebaran .................................................................. 7
2.1.5 Manfaat dan Aktivitas Farmakologis ...................................................... 8
2.1.6 Kandungan Senyawa Kimia .................................................................... 8
2.2 Tinjauan Tentang Kanker .................................................................................. 9
2.2.1 Definisi .................................................................................................... 9
2.2.2 Siklus Sel .............................................................................................. 10
2.2.3 Proses Karsinogenesis ........................................................................... 11
2.3 Tinjauan Tentang Kanker Payudara ................................................................ 12
2.3.1 Definisi .................................................................................................. 13
2.3.2 Stadium ................................................................................................. 13
2.3.3 Penyebab dan Faktor Resiko ................................................................. 19
2.3.4 Gejala Kanker Payudara........................................................................ 21
2.4 Tinjauan Tentang Antikanker ......................................................................... 22
2.4.1 Tinjauan Tentang Doxorubicin ............................................................. 24
2.4.2 Tinjauan Senyawa Metabolit Sekunder Sebagai Antikanker ................ 25
2.5 Tinjauan Tentang Sel Vero ............................................................................. 27
2.7 Tinjauan Tentang Sel Kanker Payudara .......................................................... 28
2.8 Tinjauan Tentang Metode Ekstraksi ............................................................... 29
2.9 Tinjauan Tentang Fraksinasi ........................................................................... 32
2.10 Tinjauan Tentang Metode MTT (Microculture Tetrazolium Salt) Assay ..... 32
BAB III KERANGKA KONSEPTUAL ............................................................ 34
BAB IV METODE PENELITIAN .................................................................... 37
4.1 Lokasi Penelitian ............................................................................................ 37
4.2 Bahan dan Alat Penelitian ............................................................................... 37
4.2.1 Bahan Uji .............................................................................................. 37
4.2.2 Bahan Penelitian ................................................................................... 37
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4.2.3 Alat-Alat Penelitian .............................................................................. 38
4.3 Variabel Penelitian .......................................................................................... 38
4.3.1 Variable Bebas ...................................................................................... 38
4.3.2 Variable Tergantung ............................................................................. 39
4.4 Metode Penelitian............................................................................................ 39
4.4.1 Rancangan Penelitian ............................................................................ 39
4.4.2 Kerangka Operasional ........................................................................... 40
4.5 Prosedure Kerja ............................................................................................... 41
4.5.1 Pembuatan Ekstrak Uji ......................................................................... 41
4.6 Pembuatan Media ............................................................................................ 41
4.7 Pembuatan Larutan Induk dan Larutan Uji ..................................................... 45
4.7.1 Pembuatan Larutan Induk ..................................................................... 45
4.7.2 Pembuatan Larutan Uji ......................................................................... 45
4.8 Pembuatan Larutan Induk dan Larutan Uji Kontrol Positif (Doxorubicin) .... 46
4.8.1 Pembuatan Larutan Uji ......................................................................... 46
4.9 Uji Sitotoksisitas dengan MTT ....................................................................... 46
4.10 Analisis Data ................................................................................................. 48
BAB V HASIL PENELITIAN ........................................................................... 50
5.1 Persiapan Sel MCF-7 ...................................................................................... 50
5.1.1 Hasil Perhitungan Sel MCF-7 ............................................................... 50
5.1.2 Kebutuhan Sel MCF-7 Yang Ditransfer Untuk Pengujian MTT .......... 50
5.1.3 Hasil Sitotoksisias dari Ekstrak Etanol Akar Jarak Merah (Jatropha
gossypifolia L.) Terhadap Sel Kanker Payudara MCF-7 dengan metode MTT
Assay .............................................................................................................. 51
5.2 Persiapan Sel MCF-7 Untuk Doxorubicin ...................................................... 51
5.2.1 Hasil Perhitungan Sel MCF-7 ............................................................... 51
xiii
5.2.2 Kebutuhan Sel MCF-7 Yang Ditransfer Untuk Pengujian MTT .......... 52
5.2.3 Hasil Sitotoksisias dari Doxorubicin Terhadap Sel Kanker Payudara
MCF-7 dengan metode MTT Assay ............................................................... 52
5.3 Persiapan Sel Vero ......................................................................................... 52
5.3.1 Hasil Perhitungan Sel Vero ................................................................... 52
5.3.2 Kebutuhan Sel MCF-7 Yang Ditransfer Untuk Pengujian MTT .......... 53
5.3.3 Hasil Sitotoksisias dari Doxorubicin Terhadap Sel Kanker Payudara
MCF-7 dengan metode MTT Assay ............................................................... 53
BAB VI PEMBAHASAN .................................................................................... 67
BAB VII KESIMPULAN DAN SARAN ........................................................... 61
7.1 Kesimpulan .................................................................................................... 61
7.2 Saran .............................................................................................................. 61
DAFTAR PUSTAKA .......................................................................................... 62
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DAFTAR TABEL
Tabel Halaman
II.1 Isolasi Senyawa Akar J. gossypifolia L. (Silva et al., 2014) ............................ 9
II.2 Perbedaan sel kanker payudara MCF-7 dan T47D......................................... 29
II.3 Indeks polaritas pelarut (Sarker et al., 2006) .................................................. 30
IV.1Kelompok perlakuan kultur sel kanker payudara (sel MCF-7) dan sel vero
dalam setiap percobaan. ................................................................................. 39
V.1 Data hasil uji sitotoksisitas fraksi EJG, doxorubicin, dan sel vero terhadap sel
kanker payudara MCF-7 dengan metode MTT assay .................................... 54
V.2 Hasil nilai IC50 fraksi etanol akar (Jatropha gossipifolia L.) dan Doxorubicin
terhadap sel kanker payudara MCF-7 dan sel vero dengan analisis data....... 56
xv
DAFTAR GAMBAR
Gambar Halaman
2.1(A) Akar J. gossypifolia L. dan (B) Daun, bunga, serta buahJ. gossypifolia L.
(Singh et al., 2013) ........................................................................................... 6
2.2Anatomi payudara (Dipiro,2011) ..................................................................... 13
2.3Tahap 0 (ASCO, 2005). ................................................................................... 14
2.4(A)Tahap IA, (B) Tahap IB prognostic grup 1, dan (C) Tahap IB prognostic
grup 2 (ASCO, 2005). .................................................................................... 15
2.5(A) Tahap IIA prognostic grup 1, (B) Tahap IIA prognostic grup 2, dan (C)
Tahap IIA prognostic grup 3 (ASCO, 2005).................................................. 16
2.6(A) Tahap IIB prognostic grup 1, dan (B) Tahap IIB prognostic grup 2 (ASCO,
2005). ............................................................................................................. 16
2.7(A) Tahap IIIA prognostic grup 1, (B) Tahap IIIA prognostic grup 2, (C)
Tahap IIIA prognostic grup 3, (D) Tahap IIIA prognostic grup 4, dan (E)
Tahap IIIA prognostic grup 5 (ASCO, 2005) ................................................ 17
2.8(A) Tahap IIIB prognostic grup 1, (B) Tahap IIIB prognostic grup 2, dan (C)
Tahap IIIB prognostic grup 3 (ASCO, 2005) ................................................ 18
2.9Tahap IIIC (ASCO, 2005). ............................................................................... 18
2.10Tahap 4 (ASCO, 2005). ................................................................................. 19
2.11 Struktur kimia doxorubicin (Pubchem, 2016) ............................................... 24
2.12Sel Vero (ATCC, 2008) ................................................................................. 27
2.13Sel MCF-7 (Biolabss, 2011) .......................................................................... 28
3.1 Skema Kerangka Operasional ......................................................................... 34
4.2 Skema Kerangka Operasional ......................................................................... 40
4.3 Perhitungan sel(CCRC, 2014) ......................................................................... 44
5.1 Kurva hubungan antara konsentrasi fraksi EJG dan doxorubicin terhadap %
sel hidup MCF-7 dan sel vero. ....................................................................... 55
xvi
DAFTAR LAMPIRAN
Lampiran Halaman
1. Daftar Riwayat Hidup ....................................................................................... 68
2. Surat Pernyataan................................................................................................ 69
3. Surat Determinasi Tanaman .............................................................................. 70
4. Sertifikat Kultur Jaringan .................................................................................. 71
5. Data Hasil Pembacaan Elisa Reader ................................................................. 72
6. Kerangka fraksinasi bertingkat n-Heksan, Etil Asetat, dan Etanol pada akar
Jatropha gossypifolia L. ................................................................................... 73
7. Hasil Screening Fitokimia Fraksi Etanol Akar J. gossypifolia L. dengan KLT
(Kromatografi Lapis Tipis) (Septiarini, 2016) ................................................. 74
8. Gambar Sitotoksik Dari Fraksi Etanol EJG Terhadap Sel Kanker Payudara
MCF-7 Dengan Metode MTT Assay ................................................................ 76
9. Gambar Sitotoksik Dari Fraksi Etanol EJG Terhadap Sel Vero Dengan Metode
MTT Assay ....................................................................................................... 77
10. Gambar Sitotoksik Dari Doxorubicin Terhadap Sel Kanker Payudara MCF-7
Dengan Metode MTT Assay ............................................................................. 78
11. Analisis Probit Fraksi Etanol EJG Untuk Sel MCF-7 ..................................... 79
12. Analisis Probit Fraksi Etanol EJG Untuk Sel Vero ........................................ 81
13. Analisis Probit Kontrol Positif Doxorubicin ................................................... 83
14. Perhitungan Data Elisa Reader Sel MCF-7 .................................................... 85
15. Perhitungan Data Elisa Reader Sel Vero ........................................................ 86
16. Perhitungan Data Elisa Reader Kontrol Positif Doxorubicin ......................... 87
17. Dokumentasi penelitian ................................................................................... 88
xvii
DAFTAR SINGKATAN
ACS = American Cancer Society
ATCC = American Type Culture Collection
BRCA = Breast Cancer Antigene
C = Celcius
cm = Centimeter
CCRC =Cancer Chemoprevention Research Center
DCIS = Ductal Carcinoma In Situ
Depkes RI = Departemen Kesehatan Republik indonesia
DES = Diethilstilbestrol
DMSO = Dimethyl Sulfoxide
DNA = Deoxyribose Nucleic Acid
EDTA = Ethylene Diamine Tetra Acid
EJG = Etanol Jatropha gossypifolia L.
FEJG = Fraksi Etanol Jatropha gossypifolia L.
ELISA = Enzyme-linked Immunosorbent Assay
FBS = Fetal Bovine Serum
FeCl3 = Besi (III) Klorida
G1 = Gap 1
G2 = Gap 2
HCl = Asam Klorida
HeLa = Henrietta Lacks
IC50 = Inhibitory Concentration 50
KLT = Kromatografi Lapis Tipis
KOH = Kalium Hidroksida
M = Mitosis
MCF-7 = Michigan Cancer Foundation-7
μg = Mikrogram
mg = Miligram
ml = Mililiter
mm = Milimeter
MK = Media Kultur
xviii
MTT = Microculture Tetrazolium Salt
NaOH = Natriom Hidroksida
NaHCO3 = Natrium Bikarbonat
NCI = National Cancer Institute
PBS = Phospat Buffer Salin
Riskesdas = Riset Kesehatan Dasar
S = Sintesis
SPSS = Statistic Program for Social Science
WHO = World Health Organization
62
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