Inhibition of JNK-1 by small interfering RNA induces apoptotic signaling in PC-3 prostate cancer cells

Parra,Eduardo

doi:10.3892/ijmm.2012.1055

Inhibition of JNK-1 by small interfering RNA induces apoptotic signaling in PC-3 prostate cancer cells

Authors:
- Eduardo Parra
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Affiliations: Biomedical Experimental Laboratory, Faculty of Sciences, University of Tarapaca, Arica, Chile
Published online on: July 5, 2012 https://doi.org/10.3892/ijmm.2012.1055
Pages: 923-930

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Abstract

Previous studies have shown that c-Jun-N-terminal kinase-1 (JNK-1) is involved in the transformation of primary fibroblasts and plays a role in tumor cell growth. A number of observations suggest that JNK-1 is a growth promoting factor in prostate cancer cells and blocking its function may induce apoptosis. To test this further, we used a small interfering RNA (siRNA) against JNK-1 mRNA that efficiently inhibits JNK-1 expression in the prostate cancer cell line, PC-3. The application of siRNA against JNK-1 decreased the expression of JNK-1 and affected the expression of p21, XIAP and Bcl-2, but had no effect on the expression of VEGF. In contrast, a control scramble siRNA did not affect the expression of the above indicated proteins. The downregulation of JNK-1 expression at both the mRNA and protein levels was detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Cell proliferation inhibition rates were determined by the MTT assay. The effect of JNK-1-siRNA on cell cycle distribution and cell apoptosis was determined by flow cytometry, DNA fragmentation and caspase activity. Our data showed that siRNA against JNK-1 mRNA, could efficiently suppress the expression of JNK-1 in PC-3 cells. After 5 days of transfection, the cell death rate was 52%, the apoptotic rate 26% and the viability rate 22%. In conclusion, downregulation of JNK-1 expression by siRNA against JNK-1 mRNA induces apoptotic signaling in prostate cancer PC-3 cells. The use of siRNA against JNK-1 as a novel approach to cancer therapy deserves further investigation.

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1.	M VermaP PatelM VermaBiomarkers in prostate cancer epidemiologyCancers337733798201110.3390/cancers304377324213111
2.	A JemalR SiegelJ XuE WardCancer statistics, 2010Cancer J Clin60277300201010.3322/caac.20073
3.	A VickersC SavageH LiljaFinasteride to prevent prostate cancer: should all men or only a high-risk subgroup be treated?J Clin Oncol2811121116201010.1200/JCO.2009.23.557220124185
4.	MS LuciaAK DarkePJ GoodmanFG La RosaHL ParnesLG FordCA Coltman JrIM ThompsonPathologic characteristics of cancers detected in The Prostate Cancer Prevention Trial: implications for prostate cancer detection and chemopreventionCancer Prev Res (Phila)3167173200810.1158/1940-6207.CAPR-08-007819138952
5.	K ShimadaM NakamuraE IshidaM KishiN KonishiRoles of p38- and c-jun NH2-terminal kinase-mediated pathways in 2-methoxyestradiol-induced p53 induction and apoptosisCarcinogenesis2410671075200310.1093/carcin/bgg05812807754
6.	Y FanH ChenB QiaoL LuoH MaOpposing effects of ERK and p38 MAP kinases on HeLa cell apoptosis induced by dipyrithioneMol Cells233038200717464209
7.	RJ DavisSignal transduction by the JNK group of MAP kinasesCell103239252200010.1016/S0092-8674(00)00116-111057897
8.	K SabapathyK HochedlingerSY NamA BauerM KarinE WagnerDistinct roles for JNK1 and JNK2 in regulating JNK activity and c-Jun-dependent cell proliferationMol Cell15713725200410.1016/j.molcel.2004.08.02815350216
9.	CY KuanDD YangDR SamantaJ RogerRJ DavisP RakicRA FlavellThe Jnk1 and Jnk2 protein kinases are required for regional specific apoptosis during early brain developmentNeuron22667676199910.1016/S0896-6273(00)80727-810230788
10.	H TsuikiM TnaniI OkamotoLC KenyonConstitutively active forms of c-Jun NH₂-terminal kinase are expressed in primary glial tumorsCancer Res63250255200312517805
11.	G LiaoQ TaoM KofronJS ChenA SchloemerRJ DavisJC HsiehC WylieJ HeasmanCY KuanJun NH₂-terminal kinase (JNK) prevents nuclear β-catenin accumulation and regulates axis formation in Xenopus embryosProc Natl Acad Sci USA10316313163182006
12.	E ParraJ FerreiraKnockdown of the c-Jun-N-terminal kinase expression by siRNA inhibits MCF-7 breast carcinoma cell lines growthOncol Rep2413391345201010.3892/or_0000099120878129
13.	F BostR McKayM BostO PotapovaNM DeanD MercolaThe Jun kinase 2 isoform is preferentially required for epidermal growth factor-induced transformation of human A549 lung carcinoma cellsMol Cell Biol19193819491999
14.	YM YangF BostW CharbonoN DeanR McKayJS RhimC DepatieD MercolaC-Jun NH₂-terminal kinase mediates proliferation and tumor growth of human prostate carcinomaClin Cancer Res93913982003
15.	J LiuA LinRole of JNK activation in apoptosis: a double-edged swordCell Res153642200510.1038/sj.cr.729026215686625
16.	NJ KennedyRJ DavisRole of JNK in tumor developmentCell Cycle2199201200312734425
17.	G PearsonF RobinsonTB GibsonB XuM KarandikarK BermanMH CobbMitogen-activated protein (MAP) kinase pathways: regulation and physiological functionsEndocr Rev22153183200111294822
18.	S ElmoreApoptosis: a review of programmed cell deathToxicol Pathol35495516200710.1080/0192623070132033717562483
19.	A AshkenaziVM DixitDeath receptors: signaling and modulationScience28113051308199810.1126/science.281.5381.13059721089
20.	FM FosterTW OwensJ Tanianis-HughesRB ClarkeK BrennanNJ BundredCH StreuliTargeting inhibitor of apoptosis proteins in combination with ErbB antagonists in breast cancerBreast Cancer Res11R41200910.1186/bcr232819563669
21.	Y WeiT FanM YuInhibitor of apoptosis proteins and apoptosisActa Biochim Biophys Sin (Shanghai)40278288200810.1111/j.1745-7270.2008.00407.x18401525
22.	NK SahZ KhanGJ KhanPS BisenStructural, functional and therapeutic biology of survivinCancer Lett244164171200610.1016/j.canlet.2006.03.00716621243
23.	J InostrozaL SaenzG CalafG CabelloE ParraRole of the phosphatase PP4 in the activation of JNK-1 in prostate carcinoma cell lines PC-3 and LNCaP resulting in increased AP-1 and EGR-1 activityBiol Res38163178200510.4067/S0716-9760200500020000616238095
24.	E ParraA OrtegaL SaenzDownregulation of Egr-1 by siRNA inhibits growth of human prostate carcinoma cell line PC-3Oncol Rep2215131518200919885607
25.	E ParraJ FerreiraA OrtegaOverexpression of EGR-1 modulates the activity of NF-κB and AP-1 in prostate carcinoma PC-3 and LNCaP cell linesInt J Oncol39345352201121617851
26.	E ParraF FerreiraL SaenzDiverse effects of siRNA-EGR-1 on NF-κB and AP-1 transcription factors in prostate carcinoma cell lines PC-3 and LNCaPInt J Mol Med288478532011
27.	OA MareninovaKF SungP HongA LugeaSJ PandolI GukovskyAS GukovskayaCell death in pancreatitis: caspases protect from necrotizing pancreatitisJ Biol Chem28133703381200610.1074/jbc.M51127620016339139
28.	DB LongleyTR WilsonM McEwanWL AllenU McDermottL GalliganPG Johnstonc-FLIP inhibits chemotherapy-induced colorectal cancer cell deathOncogene25838848200610.1038/sj.onc.120912216247474
29.	M KingM Radicchi-MastroianniEffects of caspase inhibition on camptothecin-induced apoptosis of HL-60 cellsCytometry492835200210.1002/cyto.1014112210608
30.	YT IpRJ DavisSignal transduction by the c-Jun N-terminal kinase (JNK) from inflamation to developmentCurr Opin Cell Biol10205219199810.1016/S0955-0674(98)80143-99561845
31.	PV MejiaJM BenitoA FernándezHD HanL MangalaC Rodríguezc-Jun-NH₂-kinase-1 inhibition leads to antitumor activity in ovarian cancerClin Cancer Res161841922010
32.	J LiuY MinemotoA Linc-Jun N-terminal protein kinase 1 (JNK1), but not JNK2, is essential for tumor necrosis factor alpha-induced c-Jun kinase activation and apoptosisMol Cell Biol241084410856200410.1128/MCB.24.24.10844-10856.200415572687
33.	LF LinczDeciphering the apoptotic pathway: all roads lead to deathImmunol Cell Biol761199810.1046/j.1440-1711.1998.00712.x9553771
34.	JM BrownLD AttardiThe role of apoptosis in cancer development and treatment responseNat Rev Cancer5231237200515738985
35.	A WicovskyN MillerN DaryabR MarienfeldC KneitzS KavuriM LeverkusB BaumannH WajantSustained JNK activation in response to tumor necrosis factor is mediated by caspases in a cell type-specific mannerJ Biol Chem28221742183200710.1074/jbc.M60616720017121845
36.	A MeriinVL GabaiJ YaglomJY VictorI ShifriniMY ShermanProteasome inhibitors activate stress kinases and induce Hsp72. Diverse effects on apoptosisJ Biol Chem27363736379199810.1074/jbc.273.11.63739497367
37.	E ParraActivation of MAP kinase family members triggered by TPA or ionomycin occurs via the protein phosphatase 4 pathway in Jurkat leukemia T cellsMol Med Rep5773778201222200849
38.	P CodognoAJ MeijerAutophagy and signaling: their role in cell survival and cell deathCell Death Differ1215091518200510.1038/sj.cdd.440175116247498
39.	M ZhaoM YangL YangY YuM XieS ZhuR KangD TangZ JiangW YuanHMGB1 regulates autophagy through increasing transcriptional activities of JNK and ERK in human myeloid leukemia cellsBMB Rep44601606201110.5483/BMBRep.2011.44.9.60121944254
40.	BK SabapathyT KallunkiJP DavidI GraefM KarinEF Wagnerc-Jun NH2-terminal kinase (JNK)1 and JNK2 have similar and stage-dependent roles in regulating T cell apoptosis and proliferationJ Exp Med193317328200110.1084/jem.193.3.31711157052
41.	C DongDD YangM WyskAJ WhitmarshRJ DavisRA FlavellDefective T cell differentiation in the absence of JNK1Science28220922095199810.1126/science.282.5396.20929851932
42.	R GjersetA HaghighiS LebedevaD MercolaGene therapy approaches to sensitization of human prostate carcinoma to cisplatin by adenoviral expression of p53 and by antisense jun kinase oligonucleotide methodsMethods Mol Biol175495520200111462854
43.	O PotapovaA HaghighiF BostC LiuMJ BirrerR GjersetD MercolaThe Jun kinase/stress-activated protein kinase pathway functions to regulate DNA repair and inhibition of the pathway sensitizes tumor cells to cisplatinJ Biol Chem2721404114044199710.1074/jbc.272.22.140419162025
44.	DL PersonsEM YazlovitskayaW CulJC PellingCisplatin-induced activation of mitogen-activated protein kinases in ovarian carcinoma cells: inhibition of extracellular signal-regulated kinase activity increases sensitivity to cisplatinClin Cancer Res5100710141999
45.	J HayakawaM OhmichiH KurachH IkegamiA KimuraT MatsuokaH JikiharaD MercolaY MurataInhibition of extracellular signal-regulated protein kinase or c-Jun N-terminal protein kinase cascade, differentially activated by cisplatin, sensitizes human ovarian cancer cell-lineJ Biol Chem2743164831654199910.1074/jbc.274.44.31648
46.	YM YangF BostW CharbonoN DeanR McKayJS RhimC DepatieD MercolaC-Jun NH₂-terminal kinase mediates proliferation and tumor growth in human prostrate carcinomaClin Cancer Res93914012003
47.	S KornblauM WombleYH QiuCE JacksonW ChenM KonoplevaEH EsteyM AndreeffSynergistic induction of apoptosis by simultaneous disruption of the Bcl-2 and MEK/MAPK pathways in acute myelogenous leukemiaBlood9934613464200210.1182/blood.V99.9.346111964319