Anticancer effects of the combined Thai noni juice

ethanolic extracts and 5‐fluorouracil against

cholangiocarcinoma cells in vitro and in vivo
Prompipak et al., 2021

Muh. Ali Khumaini M.B.

6110730013 1
Introduction
Treatment
• Surgical resection (early stage)
- Less likely to be diagnosed.

• Chemoteraphy (advanced stage)

- 5-fluorouracil (5-FU), cisplatin (CIS), and gemcitabine
(GEM) 5.

2
Objectives

Thai noni juice cholangiocarcinoma cells

extract

5‐fluorouracil
nude mouse xenograft model 3
 Methods

1. Cell culture 2. Thai noni juice extract

Cell line Name Medium Thai noni juice Chiangrai (TNJ-

Cr) and Buasri (TNJ-Bs)
cholangiocarcinoma KKU-100 RPMI-1640
cholangiocarcinoma KKU-213B RPMI-1640
normal human H69 DMEM
cholangiocyte
Extracted with absolute ethanol
(7:3) and the extract freeze dried

4
 3. Cell viability assay 4. Cell cycle analysis

8 × 103 cells/well 1 × 106 cells cultured for 24 h

Single agent or combination of Single agent or combination

5-FU & TNJ (synergistic) of 5-FU & TNJ (48
h)

24, 48, and 72 h incubation Cells harvested & washed with

cold PBS + 1% glucose and Flow cytometer
fixed in 70% ethanol for 1 h on
2 h incubation with ice
medium+MTT
washed with PBS then incubated Stained through incubation with
in RNase A 30 min Propidium Iodide (45 min)
Added DMSO and measure
absorbance

5
5. Apoptosis detection by flow cytometry

1 × 106 cells cultured for 24 h

Single agent or combination

(synergistic) of 5-FU & TNJ (48
h)

Cells harvested & washed with

cold PBS Flow cytometer

Resuspended in Annexin V- Stained with annexin V-FITC

binding buffer and PI (15 min)

6
6. Anti‑CCA in nude mice xenograft
model
i.p. administration single agent
Tumor or combination (synergistic) of
5-FU & TNJ (14 days)

Organ anatomy and morphology

+ % tumor inhibition

administered subcutaneously into the right axillary

with KKU-100 cells (2 × 106 cells in 0.1 mL of
serum-free medium mixed with Matrigel)

7
 Results
1. Antiproliferative effects of
the single agent treatment

8
2. Antiproliferative effects of
the combination treatment

9
3. Effects on cell cycle distribution

10
4. Effects on cell apoptosis induction

11
5. Antitumor effect on CCA xenograft mice

12
5. Histological examonatio on CCA xenograft mice

13
 Conclusion
Combination treatments of 5-FU and TNJ-Cr or TNJ-Bs ethanolic extracts
showed the synergistic antitumor effects on the human CCA cell lines, KKU-
100 and KKU-213B. Moreover, the combined treatment of 5-FU and TNJ-Bs
ethanolic extract was not toxic to the non-cancer H69 cells in vitro.
Additionally, TNJ ethanolic extracts enhanced the antitumor growth of 5-FU
on the mouse KKU-100 xenograft model through apoptosis induction.

14