Type conversion of secretomes in a 3D TAM2 and HCC cell co-culture system and functional importance of CXCL2 in HCC.
Macrophages play important roles in the tumor microenvironment, driving cancer progression and metastasis, particularly in hepatocellular carcinoma (HCC). However, few studies have assessed the exact secretome composition in HCC. In the present study, the impact of different phenotype of macrophages on HCC cells was investigated. Alternatively activated macrophages (M2) were found to significantly increase the proliferation, migration, and invasion abilities of SMMC7721 cells (all P < 0.05). M2 were then co-cultured with SMMC7721 cells to reconstruct the tumor microenvironment. Conditioned medium from 3D single cultures of M2, SMMC7721 cells, and their co-culture system were analyzed using quantitative proteomics via iTRAQ labeling combined with mass spectrometric analysis. Secretome analysis revealed a total of 159 differential secreted proteins in the co-culture system compared to the single culture systems, with 63 being up-regulated (>1.3-fold) and 96 down-regulated (<0.7-fold). CXCL2 was confirmed to have higher expression in the co-culture system and HCC tissues, and was selected for further investigation. Functional effects data suggested that recombinant human CXCL2 significantly enhanced the migration, invasion ability of SMMC7721 cells, and weakened adhesion ability. While CXCL2 neutralization and CXCR2 blockage significantly inhibited the effects of CXCL2 on SMMC7721 cells, indicating that CXCL2 may play pivotal role in HCC metastasis.
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Yu
Y
Lu
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Shan
S
Li
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Liping
L
Ma
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Yan
Y
Li
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Xiaolian
X
Zhang
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Qiliu
Q
Peng
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Cuiju
C
Mo
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Li
L
Huang
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China.
Xue
X
Qin
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai 200032, China.|Cancer Research Center, Institute of Biomedical Science, Fudan University, Shanghai 200032, China.
Yinkun
Y
Liu
D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D054426:Chemokine CXCL2 / Q000378:metabolism*; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D008264:Macrophages / Q000502:physiology*; D020543:Proteome / Q000032:analysis; D014407:Tumor Cells, Cultured
C499706:CXCL2 protein, human; D054426:Chemokine CXCL2; D017077:Culture Media, Conditioned; D020543:Proteome