Abstract
Purpose: To investigate the effects of chlorophenyl-benzoxime (CPBZX) on pancreatic cancer (PC) cell proliferation, invasion and migration, and the underlying mechanism of action.
 Methods: Pancreatic carcinoma cell lines (HuP-T4, HuP-T3 and BxPC-3) were cultured in Dulbecco's Modified Eagle medium (DMEM) containing 10 % fetal bovine serum (FBS), penicillin (100 U/mL) and streptomycin (10 μg/mL) at 37 ˚C in a humidified atmosphere containing 5 % CO2 and 95 % air. Cell proliferation was assessed using MTT assay. Real-time quantitative polymerase chain reaction (qRTPCR) and Western blotting were employed for the determination of changes in the levels of expression of carcinoembryonic antigen (CEA), interleukin-8 (IL-8) and cyclooxygenase-2 (COX 2). Cell invasion and migration were determined using Transwell and wound healing assays, respectively.
 Results: The results of MTT assay showed that CPBZX significantly and dose-dependently inhibited the proliferation of PC cells (p < 0.05). Incubation of HuP-T4 cells with CPBZX significantly and dosedependently reduced the invasive ability of the cells (p < 0.05). The migratory ability of HuP-T4 cells was also significantly and dose-dependently inhibited by CPBZX (p < 0.05). The results of Western blotting and qRT PCR showed that CPBZX treatment significantly and dose-dependently upregulated CEA mRNA expression (p < 0.05). On the other hand, the expressions of IL-8 and COX-2 were significantly and dose-dependently down-regulated by CPBZX. Treatment of pancreatic tumor mice with CPBZX significantly decreased tumor growth and metastasis of tumor cells to the pulmonary tissues, liver and lymph nodes (p < 0.05).
 Conclusion: The results of this study suggest that CPBZX inhibits the development and metastasis of PC via the down-regulation of IL-8 and COX 2 expressions, and therefore may find application in pancreatic cancer therapy.
Highlights
The incidence of pancreatic cancer (PC) is about 12 cases per 100,000 people in the United States of America, and approximately 6 patients in every 100,000 people throughout the world [1,2]
The present study investigated the effects of CPBZX on PC cell proliferation, invasion and migration, and the underlying mechanism
CPBZX significantly and dose-dependently inhibited the proliferation of PC cells (p < 0.05). It had no significant effect on the proliferation of normal pancreatic cells (CRL-4039) (p > 0.05)
Summary
The incidence of PC is about 12 cases per 100,000 people in the United States of America, and approximately 6 patients in every 100,000 people throughout the world [1,2]. CPBZX (2 – 10 μmol/L) was added to the cells and incubated for 2 days at 37 oC. The cells were cultured in Dulbecco's Modified Eagle medium (DMEM) containing 10 % fetal bovine serum (FBS), penicillin (100 U/mL) and. This was used to assess the migratory ability of the cells. The cells were incubated in DMEM containing 2 % FBS and treated with varied concentrations of CPBZX (2 - 10 μmol/L). Trizol RNA extraction reagent was used to extract total RNA from HuP-T4 cells after 48 h of incubation with varied doses of CPBZX (2 - 10 μmol/L). Values of p < 0.05 were considered statistically significant
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