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.

38

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42

Lampiran 1. Hasil Determinasi Tanaman Sirsak (Annona muricata L.)

43

Lampiran 1. Lanjutan

44

Lampiran 1. Lanjutan

45

Lampiran 2. Surat Keterangan telah Melaksanakan Penelitian di Laboratorium

Parasitologi Fakultas Kedokteran Universitas Gadjah Mada

Yogyakarta.

46

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

47

(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

48

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 %

49

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)

51

Lampiran 5. Lanjutan

(d)

2,5 RPMI

(a) (b) (c)

Keterangan konsentrasi: (a) 1 IC50 5ml RPMI+13 µl EEBS; (b) ½ IC50 2,5ml

RPMI; (c) ¼ IC50 2,5 ml RPMI; (d) EEBS 13 µl