Cysteamine Suppresses Invasion, Metastasis and Prolongs Survival by Inhibiting Matrix Metalloproteinases in a Mouse Model of Human Pancreatic Cancer
Figure 2
Cysteamine inhibited MMP activity in pancreatic cancer cells.
A. To measure total MMPs activity in pancreatic cancer cell lines, cells were incubated with 0–5 mM of cysteamine for 24 hours, total protein extracted and the activity measured using fluorogenic substrate. The effect of cysteamine on MMP-9 at protein level by ELISA (B), MMP-9, MMP-12 and MMP-14 mRNA levels (C) and gelatinase zymographic activities (D) was determined after incubating pancreatic cancer cells with cysteamine for 24 hours. MMP-9 activity was determined by ELISA and mRNA by qRT-PCR. β-actin was used for a reference gene in qRT-PCR. Data represent MMP-9 protein in pg/μg total cell lysate and RFU percent ratio of mRNA/ β -actin. Data are expressed as mean ± S.D. of triplicate determinations and experiments were repeated three times. Statistical significances are shown by *: P<0.05, †: p<0.01 and ‡: P<0.001.