A novel in vitro pancreatic carcinogenesis model.
Environmental factors (e.g., BaP) have been pointed out as one of the etiologies of pancreatic cancer. However, very limited experimental assays are available to identify pancreatic specific environmental mutagens or susceptibility genes. In this study, we have developed a simple in vitro cell culture model system that can be used to study the molecular and biochemical aspects of carcinogenesis in a near-normal immortalized pancreatic ductal epithelial cell lines. In order to demonstrate that xenobiotic stress response is intact in these cells, we employed standard molecular biology techniques. For examples, luciferase reporter and/or real-time quantitative PCR assays were used to determine stress-induced CYP1A1 and CYP1B1 gene expression. Western blotting and immunocytochemistry assays were used to demonstrate that TCDD or BaP could activate AhR signaling. For exploring the carcinogenesis mechanism, we incubated cells with [³H]BaP and determined BaP-DNA binding activity by measuring its radioactivity. BaP-DNA adduct formation was further confirmed by [³²P]-postlabeling assay. Finally, we demonstrated the effects of endogenous AhR or BRCA1 in BaP-DNA adduct accumulation in our cell system. As results, no apparent BaP-DNA adduct accumulation by [³²P]-postlabeling assay was found in either control-siRNA or AhR-siRNA pretreated cells. On the other hand, a significant increase of BaP-DNA adduct accumulation was found in BRCA1 knockdown cells. In conclusion, we suggest that this in vitro model may provide the feasibility for future studies on the molecular basis of pancreatic ductal cell carcinogenesis caused by dietary mutagens.
Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, Washington, DC 20057-1469, USA.
Hyo Jin
HJ
Kang
Young Bin
YB
Hong
Hee Jeong
HJ
Kim
Yong Weon
YW
Yi
Raghu G
RG
Nath
Young Soo
YS
Chang
Ho-Chan
HC
Cho
Insoo
I
Bae
D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism; D019313:BRCA1 Protein / Q000235:genetics / Q000378:metabolism; D001564:Benzo(a)pyrene; D015197:Carcinogenicity Tests; D002273:Carcinogens / Q000633:toxicity; D002460:Cell Line; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D065633:Cytochrome P-450 CYP1B1; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity; D018336:Receptors, Aryl Hydrocarbon / Q000037:antagonists & inhibitors / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D015262:Xenobiotics / Q000633:toxicity
D019313:BRCA1 Protein; D002273:Carcinogens; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D012097:Repressor Proteins; D015262:Xenobiotics; D001564:Benzo(a)pyrene; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1
Best Pract Res Clin Gastroenterol. 2006 Apr;20(2):197-209
16549324
Cancer Res. 2005 Jun 15;65(12):5045-53
15958547
Environ Mol Mutagen. 2002;40(2):93-100
12203401
Cancer Chemother Pharmacol. 2008 Nov;62(6):965-75
18259752
Carcinogenesis. 2002 Jul;23(7):1199-207
12117779
Cancer Res. 2004 Nov 1;64(21):7893-909
15520196
Carcinogenesis. 2008 Mar;29(3):656-65
18204078
Mol Pharmacol. 2004 May;65(5):1225-37
15102951
Cancer Res. 1994 Jun 1;54(11):3025-33
8187092
Int J Cancer. 2003 Jan 1;103(1):5-11
12455047
J Natl Cancer Inst. 1994 Oct 19;86(20):1510-6
7932805
Life Sci. 2004 Jan 9;74(8):1039-49
14672759
Cancer Genet Cytogenet. 2005 Apr 15;158(2):119-25
15796958
Cancer Causes Control. 2002 Apr;13(3):249-54
12020106
Cancer Res. 2004 Jul 15;64(14):4707-10
15256435
Nat Genet. 1994 Sep;8(1):27-32
7726912
Annu Rev Pharmacol Toxicol. 1995;35:307-40
7598497
Hum Genet. 2008 Oct;124(3):271-8
18762988
Natl Toxicol Program Tech Rep Ser. 2006 Sep;(526):1-180
17342195
Nat Genet. 2002 Sep;32(1):180-4
12195423
Nat Protoc. 2007;2(11):2772-81
18007613
Annu Rev Pharmacol Toxicol. 2003;43:309-34
12540743
Mol Pharmacol. 1993 Nov;44(5):911-7
8246913
Cancer Chemother Pharmacol. 2008 Sep;62(4):689-97
18074135
PLoS One. 2009;4(3):e4812
19277124
J Biol Chem. 2006 May 12;281(19):13047-13056
16543242
J Natl Cancer Inst. 1994 Jan 5;86(1):2-4
8271277
Pancreatology. 2008;8(2):110-25
18382097
Br J Cancer. 2005 Jun 6;92(11):2076-83
15886696
Annu Rev Pharmacol Toxicol. 1999;39:103-25
10331078
MedGenMed. 2005 Jun 29;7(2):60
16369438
J Biol Chem. 2006 May 26;281(21):14654-62
16567799
Cell. 1988 May 20;53(4):549-54
2453289
J Hum Genet. 2001;46(6):342-6
11393538
JOP. 2007 May 09;8(3):279-88
17495356
Am J Pathol. 1996 Jun;148(6):1763-70
8669463
J Biol Chem. 1999 Oct 1;274(40):28708-15
10497241
Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9990-5
12107286
Curr Opin Cell Biol. 2003 Jun;15(3):345-50
12787778
Drug Metab Dispos. 1974 Nov-Dec;2(6):583-8
4155680
JAMA. 2001 Aug 22-29;286(8):921-9
11509056
Drug Metab Pharmacokinet. 2006 Dec;21(6):437-57
17220560
CA Cancer J Clin. 2008 Mar-Apr;58(2):71-96
18287387
J Clin Invest. 2008 Feb;118(2):640-50
18172554
JOP. 2008 May 08;9(3):251-66
18469437
Arch Pharm Res. 2005 Mar;28(3):249-68
15832810
Am J Pathol. 2000 Nov;157(5):1623-31
11073822
Mol Cell Biol. 2009 Jul;29(13):3465-77
19380484
Science. 1994 Oct 7;266(5182):66-71
7545954
J Toxicol Environ Health A. 2005 Nov 12;68(21):1833-40
16207632
Clin Cancer Res. 2004 Jun 1;10(11):3780-7
15173085
Cancer Res. 1996 Dec 1;56(23):5360-4
8968085
Food Chem Toxicol. 1993 Sep;31(9):655-75
8406243
Proc Natl Acad Sci U S A. 2000 Jan 18;97(2):779-82
10639156
J Natl Cancer Inst. 2002 Sep 4;94(17):1293-300
12208894
Anal Biochem. 2006 Jan 1;348(1):15-23
16289438