PMID- 32884006
OWN - NLM
STAT- MEDLINE
DCOM- 20210325
LR  - 20210903
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 10
IP  - 1
DP  - 2020 Sep 3
TI  - Generation and characterization of CRISPR/Cas9-mediated MEN1 knockout BON1 cells: 
      a human pancreatic neuroendocrine cell line.
PG  - 14572
LID - 10.1038/s41598-020-71516-7 [doi]
LID - 14572
AB  - Among patients with the rare diagnosis of pancreatic neuroendocrine tumor 
      (P-NET), a substantial proportion suffer from the inherited cancer syndrome 
      multiple endocrine neoplasia type 1 (MEN1), which is caused by germline mutations 
      of the MEN1 suppressor gene. Somatic mutations and loss of the MEN1 protein 
      (menin) are frequently also found in sporadic P-NETs. Thus, a human 
      neuroendocrine pancreatic cell line with biallelic inactivation of MEN1 might be 
      of value for studying tumorigenesis. We used the polyclonal human P-NET cell line 
      BON1, which expresses menin, serotonin, chromogranin A and neurotensin, to 
      generate a monoclonal stable MEN1 knockout BON1 cell line (MEN1-KO-BON1) by 
      CRISPR/Cas9 editing. Changes in morphology, hormone secretion, and proliferation 
      were analyzed, and proteomics were assessed using nanoLC-MS/MS and Ingenuity 
      Pathway Analysis (IPA). The menin-lacking MEN1-KO-BON1 cells had increased 
      chromogranin A production and were smaller, more homogenous, rounder and grew 
      faster than their control counterparts. Proteomic analysis revealed 457 
      significantly altered proteins, and IPA identified biological functions related 
      to cancer, e.g., posttranslational modification and cell death/survival. Among 39 
      proteins with at least a two-fold difference in expression, twelve are relevant 
      in glucose homeostasis and insulin resistance. The stable monoclonal MEN1-KO-BON1 
      cell line was found to have preserved neuroendocrine differentiation, increased 
      proliferation, and an altered protein profile.
FAU - Monazzam, Azita
AU  - Monazzam A
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Li, Su-Chen
AU  - Li SC
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Wargelius, Hanna
AU  - Wargelius H
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Razmara, Masoud
AU  - Razmara M
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Bajic, Duska
AU  - Bajic D
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Mi, Jia
AU  - Mi J
AD  - Precision Medicine, BinZhou Medical University, Yantai, China.
FAU - Bergquist, Jonas
AU  - Bergquist J
AUID- ORCID: 0000-0002-4597-041X
AD  - Precision Medicine, BinZhou Medical University, Yantai, China.
AD  - Department of Chemistry - BMC, Analytical Chemistry and Neurochemistry, Uppsala 
      University, Uppsala, Sweden.
FAU - Crona, Joakim
AU  - Crona J
AUID- ORCID: 0000-0003-0677-4894
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden.
FAU - Skogseid, Britt
AU  - Skogseid B
AD  - Department of Medical Sciences, University Hospital, Uppsala University, 751 85, 
      Uppsala, Sweden. britt.skogseid@medsci.uu.se.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200903
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Biomarkers, Tumor)
RN  - 0 (MEN1 protein, human)
RN  - 0 (Proteome)
RN  - 0 (Proto-Oncogene Proteins)
SB  - IM
MH  - Biomarkers, Tumor/*metabolism
MH  - *CRISPR-Cas Systems
MH  - *Gene Editing
MH  - Gene Expression Regulation, Neoplastic
MH  - Humans
MH  - Neuroendocrine Tumors/metabolism/*pathology
MH  - Pancreatic Neoplasms/metabolism/*pathology
MH  - Proteome/*analysis
MH  - Proto-Oncogene Proteins/*antagonists & inhibitors/genetics
MH  - Tumor Cells, Cultured
PMC - PMC7471701
COIS- JC received lecture honoraria from Novartis and educational honoraria from 
      NETConnect (IPSEN). The other authors declare no conflicts of interest.
EDAT- 2020/09/05 06:00
MHDA- 2021/03/26 06:00
PMCR- 2020/09/03
CRDT- 2020/09/05 06:00
PHST- 2019/10/29 00:00 [received]
PHST- 2020/08/17 00:00 [accepted]
PHST- 2020/09/05 06:00 [entrez]
PHST- 2020/09/05 06:00 [pubmed]
PHST- 2021/03/26 06:00 [medline]
PHST- 2020/09/03 00:00 [pmc-release]
AID - 10.1038/s41598-020-71516-7 [pii]
AID - 71516 [pii]
AID - 10.1038/s41598-020-71516-7 [doi]
PST - epublish
SO  - Sci Rep. 2020 Sep 3;10(1):14572. doi: 10.1038/s41598-020-71516-7.