Tumor-derived trypsin enhances proliferation of intrahepatic cholangiocarcinoma cells by activating protease-activated receptor-2.
In primary malignant liver tumors, trypsinogen-immunoreactivity was present in 70% of intrahepatic cholangiocarcinoma (ICC) specimens, but absent in hepatocellular carcinoma (HCC) specimens. We suggest the secretion of trypsinogen to be a key difference in biological behavior between ICC and HCC cells. The purpose of this study was to investigate the secretion of tumor-derived trypsin and the expression of its specific receptor, protease-activated receptor-2 (PAR-2), in ICC using cell lines and surgical specimens. The expression of trypsinogen-1 mRNA was observed in three of four ICC cell lines, but none of three HCC cell lines. Western blot analysis detected trypsinogen-1 in serum-free conditioned medium from one of the ICC cell lines positive for the mRNA. Gelatin zymography revealed a gelatinolytic activity for trypsin, the activated form of trypsinogen, in the same conditioned medium. PAR-2 mRNA and protein were observed in ICC cell lines. The proliferative activity of ICC cells was increased by concentrations of trypsin as low as 10 nM, and peaked at 100 nM. The effect of trypsin was suppressed by a serine protease inhibitor, gabexate mesilate. PAR-2 expression was detected in 64% of ICC surgical specimens immunohistochemically. In addition, stroma fibroblasts expressed PAR-2 in 52% of ICC specimens. These results suggest that trypsinogen-1 contributes to the growth of ICC cells and also tumor-associated fibroblasts.
Nakanuma
Shin-ichi
S
Department of Gastroenterologic Surgery, Kanazawa University Graduate School of Medical Science, Kanazawa 920-8641, Japan. nakanuma@kanazawa-shaho.com
Tajima
Hidehiro
H
Okamoto
Kouichi
K
Hayashi
Hironori
H
Nakagawara
Hisatoshi
H
Onishi
Ichiro
I
Takamura
Hiroyuki
H
Kitagawa
Hirohisa
H
Fushida
Sachio
S
Tani
Takashi
T
Fujimura
Takashi
T
Kayahara
Masato
M
Ohta
Tetsuo
T
Wakayama
Tomohiko
T
Iseki
Shoichi
S
Harada
Shin-ichi
S
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001650:Bile Duct Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D001653:Bile Ducts, Intrahepatic / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D018281:Cholangiocarcinoma / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D016895:Culture Media, Serum-Free / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology; D016670:Gabexate / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D044464:Receptor, PAR-2 / Q000235:genetics / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014357:Trypsin / Q000235:genetics / Q000378:metabolism*
D016895:Culture Media, Serum-Free; D012333:RNA, Messenger; D044464:Receptor, PAR-2; D015842:Serine Proteinase Inhibitors; D016670:Gabexate; C489303:PRSS1 protein, human; D014357:Trypsin