Lipocalin-2 induces apoptosis in human hepatocellular carcinoma cells through activation of mitochondria pathways.
Lipocalin 2 (LCN2) is a secreted, iron-binding glycoprotein that is abnormally expressed in some malignant human cancers. However, the roles of LCN2 in hepatocellular carcinoma (HCC) cells are unknown. In this study, we suggested the LCN2 and LCN2R were weak detected in the HCC cell lines, LCN2 and LCN2R were found to be down-regulated in tumor tissues in 16 HCC patients. MTT, DAPI, TUNEL, and flow cytometry analyses revealed that LCN2 overexpression dramatically inhibited cell viability, induced apoptosis features of cell-cycle arrest in sub-G1 phase, in DNA fragmentation, and in condensation of chromatin in Huh-7 and SK-Hep-1 cells. Western blots were used to detect the activation of caspase, pro-apoptosis, and anti-apoptosis protein expression in overexpress-LCN2 HCC cells. LCN2-induced apoptosis was characterized by cleavage of caspase-9, -8, -3, and PARP protein, and a reduction in the mitochondrial membrane potential (MMP). Furthermore, LCN2 also enhanced the down-regulated Bcl-2 and up-regulated the expression of Bax. In addition, our experiments with caspase inhibitors LEHD-FMK and IETD-FMK prevent LCN2-induced apoptosis. We also demonstrated that treatment of overexpress-LCN2 HCC cells with the LCN2 neutralized antibody also significantly attenuated LCN2-induced cell apoptosis. These findings indicate that LCN2 overexpression can effectively induce apoptosis of HCC cells and may be used as a potent therapy against human HCC.
Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan.
Ming-Hsien
MH
Chien
Tsung-Ho
TH
Ying
Shun-Fa
SF
Yang
Ji-Kuen
JK
Yu
Chih-Wei
CW
Hsu
Shu-Ching
SC
Hsieh
Yi-Hsien
YH
Hsieh
D000209:Acute-Phase Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D057134:Antibodies, Neutralizing / Q000276:immunology; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D053938:DNA Fragmentation; D004789:Enzyme Activation; D059585:G1 Phase Cell Cycle Checkpoints; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D000071068:Lipocalin-2; D054834:Lipocalins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008113:Liver Neoplasms / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism* / Q000473:pathology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000209:Acute-Phase Proteins; D057134:Antibodies, Neutralizing; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D011518:Proto-Oncogene Proteins; D020169:Caspases