Blockade of proliferation and migration of gastric cancer  via targeting CDH17 with an artiﬁcial microRNA.
Liver-intestine cadherin (CDH17) is a novel member of the cadherin superfamily implicated in gastric cancer progression. To determine the role of CDH17 in the process of gastric cancer invasive growth, in the present study, RNA interference mediated by recombinant lentivirus vectors expressing artificial CDH17 miRNA was applied to induce a long-lasting down-regulation of CDH17 gene expression in BGC823 cells. The expression levels of CDH17, tumor cell motility, migration potential, and pro-liferation were measured by flow cytometry, real-time RT-PCR, Western blot analysis, immunofluorescence staining, wound healing assay, and MTT assay, respectively. Results show that four recombinant plasmid expression vectors encoding pre-miRNA against CDH17, pcDNA-CDH17-miR-SR1, -SR2, -SR3, and -SR4 were constructed correctly and down-regulated the CDH17 mRNA levels by 5.5, 57, 91, and 98%, respectively, in BGC823 cells which had an overexpression of CDH17. We packaged the recombinant lentiviral vector for CDH17 RNA interference with pcDNA-CDH17-miR-SR4 which had the highest interfering efficiency and succeeded in construction of the stable transfectants. Of note, more than 90% knockdown of CDH17 expression in BGC823 cells was obtained by miRNA technique. The CDH17-miRNA-transfected cells showed significant decrease in cell proliferation, cell motility, and migration in comparison with the control cells. Thus, we proposed that CDH17 may be an oncogene up-regulating invasive features of gastric cancer cells and could be a hopeful target for the control of gastric cancer progression.
Zhang
Jin
J
Department of Gastroenterology, Renmin Hospital, Wuhan University, Wuhan, Hubei Province, China.
Liu
Qi-Sheng
QS
Dong
Wei-Guo
WG
D001483:Base Sequence; D015153:Blotting, Western; D015820:Cadherins / Q000037:antagonists & inhibitors* / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D049109:Cell Proliferation; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*
C490584:CDH17 protein, human; D015820:Cadherins; D035683:MicroRNAs
Nucleic Acids Res. 2004 Feb 13;32(3):1154-8
14966264
Dig Dis Sci. 2007 Feb;52(2):536-42
17226075
RNA. 2002 Jun;8(6):842-50
12088155
Curr Opin Cell Biol. 1993 Oct;5(5):806-11
8240824
Dig Dis Sci. 2008 Jul;53(7):1811-7
17999183
Cancer Sci. 2003 Jul;94(7):575-81
12841864
Nat Genet. 2005 Nov;37(11):1289-95
16200064
Annu Rev Biochem. 1990;59:237-52
2197976
Int J Oncol. 2010 Jan;36(1):245-54
19956853
Nature. 2004 Sep 16;431(7006):350-5
15372042
Clin Cancer Res. 2006 Sep 1;12(17):5248-52
16951245
Ann Surg Oncol. 2007 May;14(5):1703-11
17211730
Gut. 2001 Jul;49(1):73-81
11413113
Mol Cell. 2002 Jun;9(6):1327-33
12086629
Science. 2004 Apr 23;304(5670):594-6
15105502
Clin Cancer Res. 2005 Jan 15;11(2 Pt 1):483-9
15701831
Proc Natl Acad Sci U S A. 2005 Sep 13;102(37):13212-7
16141338
Cancer Sci. 2003 May;94(5):425-30
12824888
J Cell Biol. 1997 Mar 10;136(5):1109-21
9060475
Biochim Biophys Acta. 1994 May 27;1198(1):11-26
8199193
Cancer. 2009 Oct 15;115(20):4753-65
19626651
Hepatology. 2009 Nov;50(5):1453-63
19676131
Cancer Res. 2008 Apr 1;68(7):2329-39
18381440
Gastroenterology. 2002 Nov;123(5):1565-77
12404231
Nat Protoc. 2007;2(2):329-33
17406593
Cancer Lett. 2004 Aug 30;212(2):253-9
15279905
Ann N Y Acad Sci. 2000;915:136-43
11193569
Dis Colon Rectum. 2007 Nov;50(11):1873-80
17828401
Biol Reprod. 2005 Sep;73(3):427-33
15901636
Methods. 2001 Dec;25(4):402-8
11846609
Biochem Biophys Res Commun. 2003 Nov 21;311(3):618-24
14623315