How the tumor microenvironment stimulates pancreaticcancer growth and progression bostonchildrens NatureComms nature
. The progression, or lack thereof, of tumors could thus potentially be explained as a competition between PRSS2 and PSAP. If tumors make more PSAP than PRSS2 they will grow more slowly, and if tumors make more PRSS2 than PSAP they will progress more rapidly. This hypothesis was born out by the observations that PRSS2 expression negatively correlates with PSAP expression in prostate cancer.
Based on the findings presented in this report, we predict that therapeutic strategies, which augment PSAP activity and inhibit PRSS2 binding to LRP1 could hold tremendous therapeutic potential for cancer patients. Significantly, this strategy should have relatively few adverse effects, as the therapeutic agents would not have direct cytotoxic activity.
WI38 cells were seeded 24 h before the treatment and synchronized for 2 h in a serum-free medium. Synchronized cells were treated with 25 µm Rac1 inhibitor NSC23766 , 10 µm JNK inhibitor III , 25μM Nutlin-3 , or PRSS2 conditioned medium for 45 min or 16 h and subjected to immunoblot analysis.. The following primary antibodies were used: Tsp-1 , LRP1 , pJNK , JNK , Phospho-c-Jun , c-Jun , β-actin , PRSS2 .
Lentiviral particles were generated by transfecting HEK293T cells and transferring the conditioned media to SUM159 or Pan02 cells, followed by selection with 1 μg/mL puromycin. Knockdown was confirmed by western blot analysis and RT-PCR.The knockdown level of PRSS2 by shRNA in Pan02 cells was measured by real-time quantitative PCR. The following primers were used: mPRSS2_F 5′-GCTCACTGCTACAAATACCGCATCCA-3′, mPRSS2_R 5′-AGGTACAGAGGCCACTCTGGCATTG-3′.
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