Ukrain affects pancreas cancer cell phenotype in vitro by targeting MMP-9 and intra-/extracellular SPARC expression.
OBJECTIVE
We investigated whether the anticancer drug Ukrain (UK) is able to modulate the expression of some of the key markers of tumor progression in pancreatic cell carcinoma, in order to assess its potential therapeutic effect.


METHODS
Three cell lines (HPAF-II, PL45, HPAC) were treated with UK (5, 10 and 20 μM) for 48 h, or left untreated. Secreted protein acidic and rich in cysteine (SPARC) mRNA levels were assessed by real-time PCR. Matrix metalloproteinases (MMP)-2 and -9 activity was analyzed by SDS zymography; SPARC protein levels in cell lysates and supernatants were determined by Western blot. Cell cycle was determined by flow cytometric analysis, and invasion by matrigel invasion assay.


RESULTS
UK down-regulated MMP-2 and MMP-9, suggesting that UK may decrease pancreatic cancer cell invasion, as confirmed by the matrigel invasion assay. SPARC protein down-regulation in supernatants points to an inhibition by UK of extracellular matrix remodeling in the tumor microenvironment. At the same time, SPARC mRNA and cellular protein level up-regulation suggests that UK can affect cell proliferation by cell cycle inhibition, showing a cell cycle G2/M arrest in UK-treated cells.


CONCLUSIONS
Our results suggest that UK modulates two major aspects involved in tumorigenesis of pancreatic cancer cells, such as extracellular matrix remodeling and cell proliferation.
Department of Surgery, University of Pisa, Pisa, Italy.
Niccola
N
Funel
Francesco
F
Costa
Letizia
L
Pettinari
Adriano
A
Taddeo
Alessandra
A
Sala
Maurizio
M
Chiriva-Internati
Everardo
E
Cobos
Graziano
G
Colombo
Aldo
A
Milzani
Daniela
D
Campani
Isabella
I
Dalle-Donne
Nicoletta
N
Gagliano
D000230:Adenocarcinoma / Q000188:drug therapy; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D001600:Berberine Alkaloids / Q000494:pharmacology* / Q000627:therapeutic use; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015536:Down-Regulation; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009361:Neoplasm Invasiveness / Q000517:prevention & control; D015676:Osteonectin / Q000096:biosynthesis*; D010190:Pancreatic Neoplasms / Q000188:drug therapy*; D010617:Phenanthridines / Q000494:pharmacology* / Q000627:therapeutic use; D012333:RNA, Messenger / Q000378:metabolism; D059016:Tumor Microenvironment / Q000187:drug effects; D015854:Up-Regulation
D000972:Antineoplastic Agents, Phytogenic; D001600:Berberine Alkaloids; D015676:Osteonectin; D010617:Phenanthridines; D012333:RNA, Messenger; C060921:ukrain; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9