Growth Inhibitory Effect of Anti-cancer drug
against Human Peritoneal Metastatic PancreaticCancer Cell Lines〇 Takashi Morikawa1), Misato Kakita1). Kana Honbori1), Miri Fumoto1),Keiko Harada1), Shigenori Enoki1), Seiichi Katayama1), Keisuke Matsuzaki2) ,Masayuki Komatsu3), Fumitaka Takeshita3), Fumiko Chiwaki3), Hiroki Sasaki3)
1) Pharmacology Department, LSI Medience Corporation
2) Ascites Treatment Center, Kanamecho Hospital
3) Biomarker Therapeutic Target Research Core, National Cancer center Research1

COI Disclosure Information
Lead Presenter/Responsible Researcher:Takashi Morikawa
I have no financial relationships to disclose.

2

Objective

Evaluations of in vitro pharmacological effects of anticancer
drugs are often conducted using cells obtained from cell banks;
however, the effects of anticancer drugs on cell lines that werepassaged for many years do not always coincide with theirclinical effects. Furthermore, the number of established cell lines
clarified as those derived from metastatic cancer is limited.
In this study, the effects of anticancer drugs were investigatedusing peritoneal metastatic pancreatic cancer cells (23 strains)that were established at the National Cancer Center (NCC) in thepast few years and passaged within 15 generations.
Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines3

Materials and Methods
Cells

Experimental design

Peritoneal metastatic pancreatic cancer cells (PMPCC) :23 strains
Pancreatic cancer cells (PCCs)(Purchased from ATCC) :MIA PaCa-2, PANC-1, HPAC and BxPC-3

Cell seeding

Culture conditions
Cells were cultivated statically in a CO2 incubator set
at 37.0℃ and 5.0% CO2 under a humidified condition.

Anticancer drugs
Cisplatina) :0.391 to 100 μM (common ratio 2),
3.91 to 1000 pM (common ratio 2),
Gemcitabinea) : 0.0000565 to 3.33 μM (common ratio 3)
Docetaxela) :a) Tokyo Chemical Industry Co., Ltd.

Evaluation of cell viability
CellTiter-Glo Luminesent Cell viability Assay(Promega K.K.)

0 hr(Day 0)

Anticancer drug
24 hr(Day 1)

Measurement
96 hr(Day 4)Cell proliferation assay was performed three times
on three separate days per cells.

Calculation of cell proliferation rate and IC50
Cell proliferation rate (%)= 100 × [(T – B4) – D1] / [(C4 – B4) – D1]

T: Luminescence intensity (LI) of anticancer drug, C4: Mean LI valueof the control group of Day 4, B4: Mean LI value of the blank groupof Day 4, B1: Mean LI value of the blank group of Day 1, C1: Mean
LI value of the control group of Day 1, D1: LI of Day 1

IC50 was calculated by using SAS 9.4 (SAS InstituteJapan Ltd., EXSUS Version 8.1.0 [CAC Croit Corporation]

Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines4Results (1)IC50 in cell growth inhibitory effects of Docetaxel to PMPCC and PCCs
(pM)

1000

5000Peritoneal metastatic
Gastric cancer cells

PMPCC

PCCs

Data represent the mean ± standard deviations. Bars without standard deviation represent duplicate data or singlicate data.

Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines

5Results (2)IC50 in cell growth inhibitory effect of Cisplatin to PMPCC and PCCs
PMPCC

PCCs

Data represent the mean ± standard deviations. Bars without standard deviation represent duplicate data or singlicate data.

Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines

6

Results (3)IC50 in cell growth inhibitory activity of Gemcitabine to PMPCC and PCCs
PMPCC

PCCs

Data represent the mean ± standard deviations. Bars without standard deviation represent duplicate data or singlicate data.

Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines

7

Results and Conclusion• An effect of docetaxel was noted in one of 23 strains, that of cisplatin wasnoted in 8 out of 23 strains, but the effect was weak in 15 strains as withdocetaxel, and that of gemcitabine was noted in 15 out of 23 strains,
but effect was weak in 7 strains.
• Comparing these results with the effect of anticancer drugs on PMPCC,
it was clarified that docetaxel has almost no effect on PMPCC.
Also, the number of PMPCC that showed an effect on gemcitabine washigher than that on cisplatin.
• An effect of docetaxel was noted in 2 of 4 PCCs, that of cisplatin wasnoted all of PCCs, and that of gemcitabine noted 3 of 4 PCCs.
It was thought that cell lines were sensitive to anticancer drugs.
We have concluded that a panel test using 23 PMPCC is useful and valid.
Growth Inhibitory Effect of Anti-cancer drug against Human Peritoneal Metastatic Pancreatic Cancer Cell Lines8