[Inhibitory effects of high mobility group box 1 antisense nucleotide on invasion of human pancreatic cancer cell line PCNA-1].
OBJECTIVE
High mobility group box 1 (HMGB1) is abundantly expressed in most of immature, and malignant cells, and is closely related to cell proliferation, invasion, and migration. We have previously proved HMGB1 over-expressed in pancreatic cancer tissues. This study was to evaluate the in vitro effects of HMGB1 antisense nucleotide on human pancreatic cancer cells invasion, and explore the underlying mechanism.


METHODS
An eukaryotic expression vector containing antisense-HMGB1 was transfected into human pancreatic cancer cell line PCNA-1. The expression of HMGB1, matrix metalloproteinase-2 (MMP-2),and MMP-9 mRNA before and after transfection was detected by reverse transcriptase-polymerase chain reaction(RT-PCR); HMGB1 protein expression was examined by Western blot. The activities of MMP-2, and MMP-9 were analyzed by gelatin zymography. The in vitro invasive ability of PCNA-1 cells was determined by Boyden chambers method.


RESULTS
Antisense-HMGB1 eukaryotic expression vector was successfully constructed. HMGB1 mRNA expression in PCNA-1 cells was effectively inhibited by antisense-HMGB1. HMGB1 protein expression was also suppressed upon transfecting (P< 0.05). Antisense-HMGB1 transfected cells showed lower MMP-2 and MMP-9 mRNA expression (P< 0.01). MMP-2 and MMP-9 activities were also inhibited by antisens-HMGB1 (P< 0.01). Cells migrating through the Boyden chamber membrane was significantly reduced as compared with the control (P< 0.01).


CONCLUSIONS
HMGB1 plays a crucial role in the invasion of pancreatic cancer, and blocking HMGB1 may be a potential strategy in preventing the migration of pancreatic cancer.
Huang
Qing-Xian
QX
Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei,430022, PR China. yhdhuang@163.com
Wang
Guo-Bin
GB
Sun
Nian-Feng
NF
Wang
Chun-You
CY
D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D024243:HMGB1 Protein / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D024243:HMGB1 Protein; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9