[The supernatant of 4T1 breast cancer cell culture increases arginase 1 content in ANA-1 macrophages].
OBJECTIVE
To explore the effect of the supernatant of 4T1 murine breast cancer cell culture on arginase 1 (Arg-1) in ANA-1 macrophages in vitro by simulating the microenvironment of breast cancer.


METHODS
The experimental ANA-1 macrophages were treated with the supernatant of 4T1 culture, and meanwhile, the control cells were cultured in the absence of the supernatant. Morphological changes of the ANA-1 macrophages were observed with a light microscope at 6, 8, 10, 24 hours, respectively. Real-time quantitative PCR (qRT-PCR) was performed to detect the levels of inducible nitric oxide synthase (iNOS) and Arg-1 mRNAs. Immunofluorescence and Western blotting were used to determine the levels of iNOS and Arg-1 proteins.


RESULTS
The qRT-PCR indicated that the level of iNOS mRNA decreased in the experiment group compared with the control group, while Arg-1 mRNA level significantly increased compared with the control group and it reached a peak at the 8th hour. The immunofluorescence and Western blotting also demonstrated that Arg-1 protein expression was enhanced in the experimental group compared with the control group. However, iNOS protein expression was no significantly different between the experiment group and the control group.


CONCLUSIONS
The supernatant of 4T1 cell culture increases Arg-1 production in ANA-1 macrophages.
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Chanjuan
C
Xie
Department of Oncology, Affiliated People's Hospital, Jiangsu University, Zhenjiang 212001, China.
Huiling
H
Shen
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Xin
X
Lin
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Yakun
Y
Lang
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Xiaolan
X
Zhu
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Tingjun
T
Bao
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Yueqin
Y
Liu
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Zhaoliang
Z
Su
Central Laboratory, Fourth Affiliated People's Hospital; Jiangsu University, Zhenjiang 212001, China.
Wenlin
W
Xu
D000818:Animals; D001119:Arginase / Q000235:genetics / Q000378:metabolism*; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D059016:Tumor Microenvironment / Q000187:drug effects
D017077:Culture Media, Conditioned; D012333:RNA, Messenger; D052247:Nitric Oxide Synthase Type II; C544448:Arg1 protein, mouse; D001119:Arginase