Establishment of primary breast cancer cell line as new model for drug screening and basic research

doi:10.19401/j.cnki.1007-3639.2024.06.004

Abstract

Abstract:

Background and purpose: In 2016 the National Cancer Institute (NCI) decided stopping to use NCI-60 cell lines for drug screening, suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research. The reason for NCI-60 cells 'retirement' was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials. Since the major cancer behaviors, such as proliferation and metastasis, are fundamentally altered with long-term culture, the tumor cell lines are not representative of the characteristics of cancer in patients. Currently, scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past. Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research. Methods: Breast cancer tissues were collected in the Department of Breast Surgery, Affiliated Hospital of Guizhou Medical University. The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University (approval number: 2022 ethics No. 313), and the collection and use of tumor tissues complied with the Declaration of Helsinki. The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium. After the cells proliferated, the media were replaced with DEME medium. Cell line STR genotyping was done to determine cell-specific genetic markers and identification. Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors. Results: We created 6 primary breast cancer cell lines. The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features, clinical diagnosis, therapeutic regimens, clinical effectiveness and prognostic outcomes. The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines. Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines. Conclusion: We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.

Key words: Breast cancer primary cell lines, Drug discovery, Basic research, New cancer model

CLC Number:

R737.9

HAO Xian, HUANG Jianjun, YANG Wenxiu, LIU Jinting, ZHANG Junhong, LUO Yubei, LI Qing, WANG Dahong, GAO Yuwei, TAN Fuyun, BO Li, ZHENG Yu, WANG Rong, FENG Jianglong, LI Jing, ZHAO Chunhua, DOU Xiaowei. Establishment of primary breast cancer cell line as new model for drug screening and basic research[J]. China Oncology, 2024, 34(6): 561-570.

Figures/Tables 8

Tab. 1

Fig.1

Tab. 2

Fig. 2

Tab. 3

Fig. 3

Fig. 4

Tab. 4

The clinical treatment data of six breast cancer patient"

Patients	BC3#	BC4#	BC5#	BC6#	BC10#	BC13#
Age/year	68	45	43	45	46	37
ER	90%	50%	90%	30%	60%	80%
PR	70%	60%	80%	10%	60%	50%
HER2	2+, FISH (-)	1+	1+	3+	1+	2+, FISH (-)
Ki-67 proliferation index	40%	30%	30%	30%	10-20%	30%
AR	80%	(-)	30%		20%	30%
TOPOⅡ-a	30%	40%	20%	40%	40%	20%
CK5/6	(-)	(+)	(-)	(-)	(-)	(-)
Menopausal status	Post-	Pre-	Pre-	Pre-	Pre-	Pre-
Specimens	Resection	Biopsy	Biopsy	Biopsy	Biopsy	Biopsy
Drug use	No	No	No	No	No	No
Tumor size D/mm	28×6×17	52×48×27	15×14×19	36×15×28	56×16×45	26×14×26
Neo/post-operative treatment	Post-operation	Neo	Neo	Neo	Neo	Neo
Neoadjuvant therapy
Treatment plan		EC-T×4	EC-T×4	EC-T×4	EC-T×6	EC-T×4
Drug use (EC×4）		Pirarubicin	Doxorubicin	Pirarubicin	Pirarubicin×6	Doxorubicin
Tumor size D/mm	28×6×17	40×33×12	19×12×15	42×11×38	39×18×33	19×11×11
Treatment effectiveness		·76.5%	·14.3%	·16.1%	·42.5%	·75.7%
Drug use (T×4)		Docetaxel	Docetaxel	Docetaxel×3	Docetaxel×2	Albumin paclitaxel×2
Tumor size D/mm		30×11×20	10×8×9	20×8×16	32×11×31	8×7×8
Treatment effectiveness compared to first diagnosis		·90.2%	·82.0%	·83.1%	·72.9%	·98.9%
Treatment effectiveness after drug use		·58.3%	·78.9%	·85.4%	·52.9%	·82.9%
pCR		G4	G3	G3	G2	G5
Neoa djuvant therapy, and therapy after operation		-	-	HP+tamoxifen	Docetaxel×2	Albumin paclitaxel×2
Adjuvant therapy after operation
Treatment plan	EC-T×4
	Epirubicin + cyclophosphamide; docetaxel
Follow-up	Whole-body bone scan suggestive of bone metastasis	Lung metastasis ER was 80%, PR was 70%, HER2 was 2+, Ki-67 proliferation index was 15%	Advise postoperative radiotherapy + oral tamoxifen, no follow-up report available	After 4 cycles of EC showing partial response, followed by 3 cycles of T with subsequent palpitations and intolerance to further treatment, surgery performed. postoperative pathology: ER 10%, PR 40%, HER2+; FISH (+), Ki-67 proliferation index was 40%, CK5/6-, TOPOⅡ-a 40%. Postoperative endocrine and targeted therapy administered. recovery good, no abnormalities on imaging.	Pleural metastasis, chest wall recurrence, thoracolumbar spine metastasis, chest mass biopsy: ER 90%, PR 80%, HER2 (2+), Ki-67 proliferation index was 50%, CK5/6 (-), No FISH performed, administered lobaplatin + capecitabine as salvage chemotherapy	Imaging showed no recurrence or metastasis

Tab. 4

References 11

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Item	BC3#	BC4#	BC5#	BC6#	BC10#	BC13#
Histological type	Invasive ductal carcinoma (40%), ductal carcinoma in situ	Invasive ductal carcinoma, ductal carcinoma in situ	Invasive ductal carcinoma	Invasive ductal carcinoma	Invasive ductal carcinoma	Invasive ductal carcinoma
Age/year	68	45	44	47	46	37
Menopausal status	Post-menopausal	Post-menopausal	Pre-menopausal	Pre-menopausal	Pre-menopausal	Pre-menopausal
Size of tumor D/mm	28×6×17	40×33×12	15×14×19	36×15×28	50×29×39	26×14×26
TNM	T2N2M0	T2N0M0	T2N0M0	T2N1M0	T4N0M0	T4N1M0
Pathological grade	Ⅲ	Ⅰ	Ⅱ	-	Ⅱ	Ⅱ
Lymph node status	Positive	Positive	Negative	Positive	Positive	Negative
Metastasis	Negative	Negative	Negative	Negative	Negative	Negative

Item	BC3#	BC4#	BC5#	BC6#	BC10#	BC13#
D19S433	12/12	12/12	12/12	12/12	12/12	12/12
D5S818	11/12	11/12	11/12	11/12	10/11/12/13	11/12
D21S11	29/29	28/29	28/29	28/29	28/29	28/29
D18S51	16/16	16/16	17/18	16/16	16/16	16/16
D6S1043	10/19	10/11/19	10/19	10/19	10/19	10/19
AMEL	XX	XX	XX	XX	XX	XX
D3S1358	14/15	14/16	14/16	14/16	14/15	14/16
D13S317	13/13	13/13	13/13	13/13	12/13	13/13
D7S820	10.1/11	10/11	10/11	10/12	9/10/11	10/12
D16S539	11/13	11/13	11/13	11/13	11/13	11/13
CSF1PO	11/11	12/12	12/12	11/12	12/13	11/12
Penta D	8/9	8/9	8/9	8/9	8/9	8/9
D2S441	11/13	11/12/13	11/13	11/13	11/13	11/13
vWA	16/17	16/16	16/16	16/16	16/16	16/16
D8S1179	15/17	15/17	15/17	17/17	15/17	17/17
TPOX	8/10	8/10	8/10	8/10	8/10	8/10
Penta E	9/12	9/13	9/13	9/13	9/13	9/13
TH01	6/6	6/6/19	6/6	6/6	6/6	6/6
D12S391	19/24	20	18/24	19/24	18/19/24	19/24
D2S1338	21/22	21/22	21/22	21/22	21/22	21/22
FGA	18/24	18/24	17/24	18/25	18/22/24	18/25

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