Download ppt - Acquire ovarian cancer cell lines from various repository

AAcquire ovarian cancer cell lines from

various repositoryATCC (6), CRT (2), DSMZ (2), ECACC

(7), Kyoto U (24), RIKEN (1)

Cell line expansion & bankingSGOCL (43)

Downstream Analysis

Immunofluorescence StainingDNA, RNA, Protein Extraction

BRNA

Gene Expression Profiling

Phenotype-specificGene Signatures

EMT RT2 Profiler qPCR Array

Phenotype-specificGene Expression

Affymetrix GeneChip Human Gene 1.0 ST Array

QA Agilent Bioanalyzer

Parametric / NonparametricAnalysis

Cell

Functional Assays

Migration AssayInvasion Assay

Anoikis AssayColony Assay

Endpoint ReadoutImage Analysis

Establish EM Phenotypic Characterisation

Validate EMT Marker Expression

Create EMT Spectrum

Functional Correlation

Supplementary Figure 1



A

Caov3 OVCAR-3PEO1 OVCAR8

OVCA433IGROV1 PEO4OVCA429

CH1 SKOV3DOV13 Hey C2

A2780 Hey A8A1847 TykNu

B

Caov3

IGROV1 OVCA433 OVCA429

CH1 DOV13 SKOV3 Hey C2

A1847

PEO1 OVCAR3 OVCAR8

A2780 Hey A8 TykNu

Migration Invasion

C D E

OV

CA

433

Hey

C2

Ep

ith

elia

lIn

term

edia

teE

Inte

rmed

iate

MM

esen

chy

mal

Invasive Front

F

SKOV3 subcutaneous tumours Day 59

SKOV3 subcutaneous tumour. Measurements

in centimeters (cm)

Control 100 nM 5 MA

ZD

0530

OV

CA

R3

(E)

500 nMA

ZD

0530

OV

CA

433

(In

t E

)

AZ

D05

30

SK

OV

3 (I

nt

M)

AZ

D05

30

OV

CA

R10

(M)



A B


A

B

Supplementary Figure 71-yr PFS 2-yr PFS 3-yr PFS 4-yr PFSA

B

Phenotype Comparison

Progression-free Survival (PFS)p-value (Log-rank)

1-yr 2-yr 3-yr 4-yrE vs IE 0.079 0.691 0.7625 0.4864E vs IM 0.21 0.163 0.1822 0.2253E vs M 0.0014 0.0002 0.0024 0.007

IE vs IM 0.9636 0.3289 0.1544 0.1043IE vs M 0.1538 0.0036 0.0053 0.0041IM vs M 0.2978 0.2209 0.4282 0.5109

CPhenotype

Comparison

Difference in RMST (95% CI)

12 month 24 month 36 month 48 month

E vs IE 0.468 (0.346 - 0.589) 0.979 (0.813 - 1.145) 0.034 (-0.125 - 0.192) -1.96 (-2.101 - -1.819)E vs IM 0.487 (0.370 - 0.604) 2.311 (2.121 - 2.501) 3.362 (3.18 - 3.544) 4.173 (4.007 - 4.338)E vs M 1.011 (0.875 - 1.147) 4.155 (3.95 - 4.36) 5.919 (5.731 - 6.106) 6.542 (6.378 - 6.705)

IE vs IM 0.019 (-0.129 - 0.167) 1.332 (1.141 - 1.524) 3.328 (3.159 - 3.497) 6.133 (5.987 - 6.278)IE vs M 0.543 (0.376 - 0.71) 3.176 (2.969 - 3.382) 5.885 (5.711 - 6.059) 8.502 (8.358 - 8.645)IM vs M 0.524 (0.362 - 0.686) 1.844 (1.614 - 2.074) 2.557 (2.359 - 2.754) 2.369 (2.201 - 2.537)


E-cad/DAPI N-Cad/DAPI