BAB IV KESIMPULAN DAN SARAN A. Kesimpulan B. Saraneprints.unwahas.ac.id/1517/5/BAB IV.pdf37 BAB IV...
Transcript of BAB IV KESIMPULAN DAN SARAN A. Kesimpulan B. Saraneprints.unwahas.ac.id/1517/5/BAB IV.pdf37 BAB IV...
37
BAB IV
KESIMPULAN DAN SARAN
A. Kesimpulan
Ekstrak Etanolik Biji Sirsak memperlihatkan adanya penghambatan
siklus sel kanker payudara T47D di fase G1 dan fase G2/M. Besarnya persentase
akumulasi sel pada fase G1 dan G2/M ini sedikit lebih tinggi bila dibandingkan
dengan sel tanpa perlakuan (kontrol sel).
B. Saran
Perlu dilakukan penelitian lebih lanjut yaitu fraksinasi ekstrak etanolik
biji sirsak terhadap sel kanker payudara T47D.
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Lampiran 2. Surat Keterangan telah Melaksanakan Penelitian di Laboratorium
Parasitologi Fakultas Kedokteran Universitas Gadjah Mada
Yogyakarta.
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Lampiran 3. Perhitungan Sel T47D, Seri Konsentrasi Ekstrak Etanolik Biji
Sirsak
1. Sel T47D
a. Perhitungan Sel
Jumlah sel terhitung = 156 x 104
b. Pembuatan Suspensi Sel (Stok)
Sel T47D untuk perlakuan = 1 x 104 sel/sumuran
Jumlah sel yang ditanam dalam setiap sumuran adalah 10.000 sel
Volume yang diambil =
= 0,33 ml + Media Kultur (RPMI) ad 5
ml.
2. Seri Konsentrasi Ekstrak Etanolik Biji Sirsak
a. Pembuatan Larutan Stok Konsentrasi 100.000 µg
/ml
Sebanyak 11 mg Ekstrak Etanolik Biji Sirsak dilarutkan dalam 110 µl
DMSO ditambahkan media ad 10 ml.
Konsentrasi stok 10 mg/100 µl = 10.000 µg/100 µl = 100.000 µg/ml.
b. Pembuatan Seri Konsntrasi
Sel T47D 3 replikasi= 300 µl, dilebihkan menjadi 400 µl
Pembuatan seri konsentrasi 1000 µg
/ml
V1 X C1 = V2 X C2
V1 X 100.000 µg
/ml = 800 µl X 1000 µg
/ml
V1 =
= 8 µl ad 392 µl MK
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(h)
MK
(a) (b) (c) (d) (e) (f) (g)
Keterangan konsentrasi: (a) 1000 ppm, 800 µl MK; (b) 500 ppm, 400 µl MK;
(c) 250 ppm, 400 µl MK; (d) 125 ppm, 400 µl MK; (e) 62,5 ppm, 400 µl MK; (f)
31, 25 ppm, 400 µl MK; (g) 15,625 ppm, 400 µl MK; (h) sampel EEBS 8 µl
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Lampiran 4. Penentuan Nilai IC50 Ekstrak Etanolik Biji Sirsak pada Sel Kanker
Payudara T47D
Konsentrasi
EEBS (µg/ml)
Absorbansi
Rata-Rata
Absorbansi
%
Viabilitas
(%) 111 22 %sel
hidup33
500
125
62.5
31.25
15.625
0.143
0.709
0.707
0.744
0.792
0.143
0.749
0.84
0.732
0.834
0.15
0.706
0.73
0.783
0.788
0.145
0.721
0.759
0.753
0.805
2.76
83.50
88.79
87.94
95.19
Kontrol Media
Kontrol Sel
0.121
0.849
0.12
0.822
0.136
0.846
0.126
0.839
Perhitungan Nilai % Viabilitas EEBS
% Viabilitas =
% Viabilitas Kons 500 =
= 2,76 %
% Viabilitas Kons 125 =
= 83,50 %
% Viabilitas Kons 62,5 =
= 88,79 %
% Viabilitas Kons 31,25=
= 87,94 %
% Viabilitas Kons 15,625=
= 95,19 %
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Lampiran 4. Lanjutan
Analisis Regresi Linier dengan Ms.Excel 2010
Y = -0,1903x + 99,593
R2
= 0,9837
Mencari IC50, sehingga Y = 50
50 = -1903x + 99,593
x =
= 260,604 µg/ml~261 µg/ml
Sehingga diperoleh IC50 Ekstrak Etanolik Biji Sirsak 261 µg/ml
y = -0.1903x + 99.593 R² = 0.9837
0
20
40
60
80
100
120
0 100 200 300 400 500 600
Konsentrasi EEBS (µg/ml)
Via
bil
itas
sel
(%
)
50
Lampiran 5. Perhitungan Sel dan Konsentrasi Ekstrak Etanolik Biji Sirsak
(EEBS) Uji Modulasi Siklus Sel
1. Sel T47D
a. Perhitungan Sel
Jumlah sel terhitung = 40x104
sel
b. Pembuatan Suspensi Sel (Stok)
Sel T47D untuk perlakuan = 10x105
sel/sumuran
Jumlah sel yang ditanam dalam setiap sumuran adalah 1.000.000 sel
Volume yang diambil= 2,5 ml
2. Perhitungan konsentrasi EEBS yang diambil
V1 X N1 = V2 X N2
5ml x 261 μg/ml = V2 X 100.000
V2 = 0,0130 ml
= 13 μl ad 5 ml Media Kultur (RPMI)