Establishment and characterization of cell lines from chromosomal instable colorectal cancer

Table 1 Clinical and pathological characteristics of patients as well as cell line establishment protocol

Tumor-ID	Age/gender	Tumor location	TNM-stage	UICC stage	Tumor type	Molecular type	β-catenin translocation	Corresponding xenograft	Direct cell line establishment	Cell line from xenograft	Paired B-LCL
HROC18	65/F	Caecum	G2T2N0 M0	I	Primary adenocarcinoma	spStd	+	-	+	-	Yes
HROC32	83/F	Ascending colon	G2T4N2 M1	IV	Primary adenocarcinoma	spStd	+	+	+	+	Yes

F: Female; spStd: Sporadic standard; B-LCL: B lymphoid cell line; +: Positive; -: Negative.

Table 2 Flow cytometric phenotyping of colorectal carcinoma cell lines and major histocompatibility complex expression with and without interferon-γ pre-treatment

Antigen	HROC18P	HROC32P
CD58	3.6%	16.4%
CD71	38.5%	39.4%
CD50	15.1%	20.3%
CD54	8.9%	27.0%
CD102	22.4%	11.8%
CD326	88.1%	83.0%
CD24	9.5%	0.6%
CD44	52.3%	41.4%
CD66acde	3.3%	13.3%
CD15	75.1%	46.7%
CD55	1.0%	6.8%
HLA-A2	98.6%	96.5%
MHC class I
- IFN-γ	99.0%	99.0%
+ IFN-γ	99.0%	99.0%
MHC class II
- IFN-γ	0.0%	2.5%
+ IFN-γ	77.0%	80.0%

Data are given from one representative experiment. CD: Cluster of differentiation; IFN: Interferon; HLA: Human leukocyte antigen; MHC: Major histocompatibility complex.

Table 3 Human leukocyte antigen: Typing of colorectal carcinoma cell lines

	HLA class I						HLA class I
DNA	A1		B1		Cw1		DRB11		DQB11
	1 allele	2 allele	1 allele	2 allele	1 allele	2 allele	1 allele	2 allele	1 allele	2 allele
HROC18	01	02	08	39	07	-	03	13	02	03
HROC32P	01	02	08	51	07	14	03	11	02	03

¹The number of the alleles. Information on both alleles is provided; in case of homozygosity the “second” allele is marked by “-“. HLA: Human leukocyte antigen.

Table 4 Comparative molecular characterization of patients’ tumor and corresponding colorectal carcinoma cell lines

		TP53				APC		KRAS
Tumor-ID		Exon 5	Exon 6	Exon 7	Exon 8	Cd1	Cd2	Cd12	Cd13	B-rafV600
HROC18	Patient tumor	wt	wt	wt	mut	mut	wt	wt	wt	wt
	Cell line	wt	wt	wt	mut	mut	wt	wt	wt	wt
HROC32	Patient tumor	wt	wt	wt	mut	wt	wt	mut	wt	wt
	P-Cell line	wt	wt	wt	mut	wt	wt	mut	wt	wt
	X-Cell line	wt	wt	wt	mut	wt	wt	mut	wt	wt

wt: Wild type; mut: Mutated; Cd: Codon.

Table 5 IC₅₀ values of antitumor drugs evaluated for colorectal carcinoma cell lines

Cell line	IC50
	5-FU(μg/mL)	Cisplatin(μg/mL)	Irinotecan(μmol/L)	Gemcitabine(μmol/L)	Paclitaxel1	Trastuzumab2	Erlotinib(μmol/L)	Nilotinib(μmol/L)	Rapamycin(μmol/L)
HROC18	0.48 ± 0.28	1.58 ± 0.29	7.88 ± 2.26	3.35 ± 1.18	Resistant	Resistant	12.21 ± 5.4	7.55 ± 1.77	0.08 ± 0.04
HROC32P	0.15 ± 0.07	0.65 ± 0.40	0.15 ± 0.07	3.90 ± 2.16	Resistant	Resistant	1.40 ± 1.01	5.47 ± 3.13	0.14 ± 0.14
HROC32X	1.97 ± 1.95	1.30 ± 0.46	3.83 ± 1.40	5.90 ± 1.06	Resistant	Resistant	0.87 ± 0.15	5.5 ± 3.1	0.13 ± 0.06

¹Range 0.9 nmol/L: 60 mmol/L;

²Range: 310 ng/mL: 20 μg/mL. Values are given as mean, resulting from at least three independent experiments each performed in triplicates. 5-FU: 5-Fluorouracil.