Cordycepin inhibits lipopolysaccharide-induced cell migration and invasion in human colorectal carcinoma HCT-116 cells through down-regulation of prostaglandin E2 receptor EP4원문보기
Jeong, Jin-Woo
(Freshwater Bioresources Utilization Bureau, Nakdonggang National Institute of Biological Resources)
,
Park, Cheol
(Department of Molecular Biology, College of Natural Sciences, Dongeui University)
,
Cha, Hee-Jae
(Department of Parasitology and Genetics, Kosin University College of Medicine)
,
Hong, Su Hyun
(Department of Biochemistry, Dong-Eui University College of Korean Medicine)
,
Park, Shin-Hyung
(Department of Pathology, Dong-Eui University College of Korean Medicine)
,
Kim, Gi-Young
(Department of Marine Life Sciences, Jeju National University)
,
Kim, Woo Jean
(Department of Anatomy, Kosin University College of Medicine)
,
Kim, Cheol Hong
(Department of Pediatrics, Sungkyunkwan University Samsung Changwon Hospital)
,
Song, Kyoung Seob
(Department of Physiology, Kosin University College of Medicine)
,
Choi, Yung Hyun
(Department of Biochemistry, Dong-Eui University College of Korean Medicine)
Prostaglandin $E_2$ ($PGE_2$), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors, including colorectal cancer. Because $PGE_2$ functions by signaling through $PGE_2$ receptors (EPs), which regul...
Prostaglandin $E_2$ ($PGE_2$), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors, including colorectal cancer. Because $PGE_2$ functions by signaling through $PGE_2$ receptors (EPs), which regulate tumor cell growth, invasion, and migration, there has been a growing amount of interest in the therapeutic potential of targeting EPs. In the present study, we investigated the role of EP4 on the effectiveness of cordycepin in inhibiting the migration and invasion of HCT116 human colorectal carcinoma cells. Our data indicate that cordycepin suppressed lipopolysaccharide (LPS)-enhanced cell migration and invasion through the inactivation of matrix metalloproteinase (MMP)-9 as well as the down-regulation of COX-2 expression and $PGE_2$ production. These events were shown to be associated with the inactivation of EP4 and activation of AMP-activated protein kinase (AMPK). Moreover, the EP4 antagonist AH23848 prevented LPS-induced MMP-9 expression and cell invasion in HCT116 cells. However, the AMPK inhibitor, compound C, as well as AMPK knockdown via siRNA, attenuated the cordycepin-induced inhibition of EP4 expression. Cordycepin treatment also reduced the activation of CREB. These findings indicate that cordycepin suppresses the migration and invasion of HCT116 cells through modulating EP4 expression and the AMPK-CREB signaling pathway. Therefore, cordycepin has the potential to serve as a potent anti-cancer agent in therapeutic strategies against colorectal cancer metastasis.
Prostaglandin $E_2$ ($PGE_2$), a major product of cyclooxygenase-2 (COX-2), plays an important role in the carcinogenesis of many solid tumors, including colorectal cancer. Because $PGE_2$ functions by signaling through $PGE_2$ receptors (EPs), which regulate tumor cell growth, invasion, and migration, there has been a growing amount of interest in the therapeutic potential of targeting EPs. In the present study, we investigated the role of EP4 on the effectiveness of cordycepin in inhibiting the migration and invasion of HCT116 human colorectal carcinoma cells. Our data indicate that cordycepin suppressed lipopolysaccharide (LPS)-enhanced cell migration and invasion through the inactivation of matrix metalloproteinase (MMP)-9 as well as the down-regulation of COX-2 expression and $PGE_2$ production. These events were shown to be associated with the inactivation of EP4 and activation of AMP-activated protein kinase (AMPK). Moreover, the EP4 antagonist AH23848 prevented LPS-induced MMP-9 expression and cell invasion in HCT116 cells. However, the AMPK inhibitor, compound C, as well as AMPK knockdown via siRNA, attenuated the cordycepin-induced inhibition of EP4 expression. Cordycepin treatment also reduced the activation of CREB. These findings indicate that cordycepin suppresses the migration and invasion of HCT116 cells through modulating EP4 expression and the AMPK-CREB signaling pathway. Therefore, cordycepin has the potential to serve as a potent anti-cancer agent in therapeutic strategies against colorectal cancer metastasis.
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문제 정의
Controlling the expression of the EP4 receptor may be a new strategy for preventing cell migration and invasion of HCT-116 cells. This study also highlights the potential for the therapeutic use of cordycepin in the treatment of colorectal cancer.
In conclusion, our results demonstrate that the levels of CREB activation were dramatically decreased by the negative regulation of AMPK expression through the deactivation of EP4 in cordycepin-treated HCT116 cells. This study provides novel insights into the molecular mechanisms of cell migration in HCT-116 cells. Controlling the expression of the EP4 receptor may be a new strategy for preventing cell migration and invasion of HCT-116 cells.
가설 설정
Because cordycepin can greatly affect the pathogenesis of colorectal disease, we hypothesized that cordycepin downregulates EP4 expression and its downstream signaling functions in human colorectal cancer cells. In order to examine the signaling pathway involved, we performed in vitro human cell-based assays.
In this study, we examined the anti-cell migration and invasion effects of cordycepin in LPS-treated human colorectal carcinoma HCT-116 cells. Cordycepin had no cytotoxic effects on HCT-116 cells at the concentrations tested (Fig.
대상 데이터
Louis, MO, USA). Antibodies against COX-2, MMP-2, MMP-9, AMPK, pAMPK, ACC, pACC, CREB, pCREB, and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The peroxidase-labeled donkey anti-rabbit immunoglobulin, peroxidase-labeled sheep anti-mouse immunoglobulin, and enhanced chemiluminescence (ECL) detection kit were purchased from Amersham Corp.
The enzyme-linked immunosorbent assay (ELISA) kit for PGE2 was obtained from R&D Systems (Minneapolis, MN, USA). COX-2, EP1-4, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) oligonucleotide primers were purchased from Bioneer (Seoul, Korea). All other chemicals were purchased from Sigma-Aldrich Chemical Co.
Cordycepin (MW, 251.2; Product No. C3394) from C. militaris, LPS (Escherichia coli 026:B6), Griess reagent, Tween-20, bovine serum albumin (BSA), H89, forskolin, compound C, and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) were purchased from SigmaAldrich Chemical Co. (St. Louis, MO, USA). Antibodies against COX-2, MMP-2, MMP-9, AMPK, pAMPK, ACC, pACC, CREB, pCREB, and actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
HCT-116 human colon cancer cells were obtained from the American Type Culture Collection (Manassas, VA, USA). These cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Gibco BRL, Gaithersburg, MD, USA) and 1% penicillin-streptomycin at 37˚C in a humid environment containing 5% CO2.
성능/효과
In conclusion, our results demonstrate that the levels of CREB activation were dramatically decreased by the negative regulation of AMPK expression through the deactivation of EP4 in cordycepin-treated HCT116 cells. This study provides novel insights into the molecular mechanisms of cell migration in HCT-116 cells.
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