Chinese Journal of Biochemistry and Molecular Biology. Effect of IL KAP on Tumorigenesis and Tumor Development

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ISSN 100727626 CN 1123870ΠQ Chinese Journal of Biochemistry and Molecular Biology IL KAP 2009 6 25 (6) :489 494 1), 2), 1), 1) 3 ( 1), 130021 ; 2), 130118) ( ILKAP) 2C( PP2C),,. ILKAP,. ILKAP,. ILKAP, c2jun Π (JNKΠMAPK).,, ILKAP, ILKAP,. ; 2C ; ; Q556 Effect of IL KAP on Tumorigenesis and Tumor Development ZHOU Wang 1), GUAN Ke2Xing 2), ZOU Xue 1), ZHANG Ying2Jiu 1) 3 ( 1) Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130021, China ; 2) College of Life Sciences, Jilin Agricultural University, Changchun 130118, China) Abstract ILKAP ( integrin2linked kinase2associated phosphatase) is a novel member of PP2C (protein phosphatase 2C) family. The results have revealed that IL KAP is an apoptosis2associated phosphatase. IL KAP is ubiquitously expressed in human tissues, especially in skeletal muscle, liver and kidney ones. IL KAP plays a major physiological role in the regulation on cellular apoptosis, and involved in the suppression of the tumorigenesis and tumor development. IL KAP played its role mainly based on the integrin and JNKΠMAPK (c2 J un N2terminal kinaseπmitogen2activated protein kinase) signaling pathways. On the other hand, the loss of heterozygosity of IL KAP gene has been confirmed in many tumor tissues. Moreover, we noted that a truncated isoform of ILKAP by Splice2Site Mutations of ILKAP gene also existed in many tumor tissues, which failed to suppress the tumorigenesis and tumor development. Key words ILKAP ; PP2C ; tumor ; apoptosis,.,,,.,,. ( integrin2linked kinase2associated phosphatase, ILKAP).,.,. ILKAP 1 (integrin2linked kinase21, ILK21), 1 (apoptosis signal2regulating kinase 1, ASK1) Thr845 JNKΠMAPK :2009202211 ; :2009203217 (No. 2006) 3 Tel : 862431288498104 ; E2mail : yingjiu @jlu. edu. cn Received : February 11,2009 ; Accepted : March 17, 2009 Supported by Project Sponsored by the Scientific Research Foundation, State Education Ministry for the Returned Overseas Chinese Scholars (No. 2006) 3 Corresponding author Tel : 862431288498104 ; E2mail : yingjiu @jlu. edu. cn

490 25..,, ILKAP,, ILKAP, ILKAP,. ILKAP ILKAP,, ILKAP ; ILKAP, N. 1 IL KAP 1998,Tong [1 ], C 2C ( protein phosphatase 2C, PP2C), PP2C PP2C 30 %, N 76., ILKAP, ILKAP, Fig11 (A) : N, PP2C. ILKAP,Tong PP2C, PP2C. 2001,Leung2Hagesteijn [2], 1(Ilk21), PP2C, PP2C Ilk21, PP2C (ILKAP).,ILKAP. Fig. 1 Structure scheme and PP2C domain of IL KAP (A) Two 2sheet lamellar form a flat lamellar space structure in the center of ILKAP, its active site is located at the top of the spatial structure of the opening ; (B) The active site of ILKAP and its mutation in tumor tissues. ILKAP mutations are concentrated in its N terminal, and can not express the activity ILKAP 2 IL KAP ILKAP PP2C, 44 kd, 392, N 76 C PP2C, Fig11 (A). Northern, ILKAP,,, [1 ]. ILKAP, 95 % 95 % 97 %, Fig12 (A). ILKAP PP2C PP2C,PP2C,PP2C PP2C 31 % 29 % 38 %,

6 : ILKAP 491 PP2C PP2C PP2C 78 %. 37 %,PP2C PP2C 37 % Fig12 (B). N 76, ILKAP., ILKAP N ILKAP,. ILKAP C PP2C, PP2C 11, ILKAP Π, Fig11 (B). Fig. 2 Homology analysis of IL KAP (A) Homology analysis of the PP2C2like domain of ILKAP among human, mouse and rat. The PP2C2like domain of ILKAP from human body share 95 % sequence identity with those from rat and mouse, while it is 97 % between rat and mouse ; (B) Homology analysis of the PP2C2like domain in ILKAP, PP2C, PP2C and PP2C. The PP2C2like domain of ILKAP share 31 %,29 % or 38 % sequence identity with the PP2C, PP2C or PP2C, respectively, and PP2C does 78 % or 37 % sequence identity with PP2C or PP2C, while it is 37 % sequence identity between PP2C and PP2C D1,. PP2C,,ILKAP Ilk21 Mg 2 + Mn 2 + [3 ], [15 ]. ILKAP, Mn 2 + Mg 2 +, Ilk21, Ilk21 [4 ]. PP2C,, Ilk21 ILKAP, [2 ]. ILKAP. Kumar [16 ] [5 ] [6., H 2 O ] 2, (LNCaP), [7 ] PP2C. ILKAP Ilk21,,ILKAP Mn 2 +, Mg 2 +, GSK23. RNA Mg 2 + [1 ]. ILKAP Mg 2 + ILKAP, cyclin D1. PP2C 3 IL KAP (integrin signaling pathway) 20.,., [8 10 ]. Ilk21. 2 B (protein kinase B,PKB) 3 (glycogen synthase kinase 3,GSK23 ). Ilk21 PKB Ser473, PKB,, [11 ]. Ilk21 GSK23 Ser9, ( 2catenin), C2Myc ( cellular myelocytomatosis oncogene,c2myc) D1 (cyclin D1), [12,13 ]., ILK21,,, ILK21., ILK21, ILK21, Ilk21 3 [14 ]. ILKAP Ilk21,, Fig13. Leung2Hagesteijn, Ilk21 ILKAP,, GSK23, 2 catenin, C2Myc ILKAP LNCaP,

492 25 Fig. 3 Apoptosis regulation of IL KAP in integrin signaling and JNK signaling pathways ILKAP activity inhibits ILK2 GSK3 signaling, suggesting that ILKAP activity inhibits oncogenic transformation. ILKAP also associated with ASK1, activating this kinase by enhancing the phosphorylation of Thr845. These observations suggest that ILKAP might act as a positive regulator of TNF 2induced apoptosis G 1, RNA ILKAP, S. ILKAP, LNCaP,. Kumar,ILKAP GSK23,, ILKAP [16 ]. 4 IL KAP JNK c2jun ( c2jun N2terminal kinase, JNK) 1990 (mitogen2activated protein kinase,mapk), Π [17 ]. JNK JNK ( ).,JNK,. JNK., JNK [18 20 ].JNK. JNKΠSAPK,. 1 (apoptosis signal2regulating kinase 1,ASK1) JNKΠSAPK. ASK1 Thr845,, JNKΠSAPK., JNKΠSAPK, ILKAP ASK1 Thr845 [21 ]. ILKAP ASK1, JNKΠSAPK,, Fig13. ASK1 Thr845, ILKAP ASK1 (Ser83 Ser967 Ser1034) [22,23 ]. 5 IL KAP,,, (loss of heterozygosity LOH), 2,. LOH,. 2007, Cengiz [24 ] 2q21237, ILKAP.,, ILKAP. ILKAP, ILKAP, ILKAP, ILKAP cdna, ILKAP. Trizol RNA, cdna, PCR, PCR pucm2t.., ILKAP, N 44, 77 ( GenBank, FJ491380) ; ILKAP, N 96

6 : ILKAP 493, 99 ( GeneBank, FJ491381), ILKAP. ILKAP,. 6, ILKAP ILK21, G 1,,,. ILKAP ASK1, JNKΠSAPK,. ILKAP 2q21237 ILKAP. ILKAP, ILKAP,, ILKAP PP2C,., ILKAP, ILKAP, ILKAP, ILKAP, ILKAP., ILKAP, ILKAP, N, ILKAP,., ILKAP,,. ( References) [ 1 ] Tong Y, Quirion R, Shen SH. Cloning and characterization of a novel mammalian PP2C isozyme [J ]. J Biol Chem,1998,273 (52) : 35282235290 [ 2 ] Leung2Hagesteijn C, Mahendra A, Naruszewicz I, et al. Modulation of integrin signal transduction by ILKAP, a protein phosphatase 2C associating with the integrin2linked kinase, ILK1 [ J ]. EMBO J, 2001,20 (9) :216022170 [ 3 ] Schweighofer A, Hirt H, Meskiene I. Plant PP2C phosphatases : emerging functions in stress signaling[j ]. Trends Plant Sci,2004, 9 (5) :2362243 [ 4 ] Cohen P T. Novel protein serineπthreonine phosphatases : variety is the spice of life[j ]. Trends Biochem Sci,1997,22 (7) : 2452251 [ 5 ] Lammers T, Lavi S. Role of type 2C protein phosphatases in growth regulation and in cellular stress signaling [J ]. Crit Rev Biochem Mol Biol,2007,42 (6) :4372461 [ 6 ] Meinhard M, Rodriguez PL, Grill E. The sensitivity of ABI2 to hydrogen peroxide links the abscisic acid2response regulator to redox signalling[j ]. Planta, 2002, 214(5) :7752782 [ 7 ] Baudouin E, Meskiene I, Hirt H. Unsaturated fatty acids inhibit MP2C, a protein phosphatase 2C involved in the wound2induced MAP kinase pathway regulation [J ]. Plant J, 1999,20(3) :3432348 [ 8 ] Dougherty GW, Jose C, Gimona M, et al. The Rsu212PINCH12ILK complex is regulated by Ras activation in tumor cells[j ]. Eur J Cell Biol,2008, 87(829) :7212734 [ 9 ] Guo W, Jiang H, Gray V, et al. Role of the integrin2linked kinase ( ILK) in determining neuronal polarity[j ]. Dev Biol,2007,306(2) : 4572468 [10 ] Zahir N, Weaver VM. Death in the third dimension : apoptosis regulation and tissue architecture[j ]. Curr Opin Genet Dev,2004,14 (1) :71280 [11 ] Chang F, Lee J T, Navolanic P M, et al. Involvement of PI3KΠAkt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy [ J ]. Leukemia, 2003, 17 (3) : 5902603 [ 12 ] Novak A, Hsu SC, Leung2Hagesteijn C, et al. Cell adhesion and the integrin2linked kinase regulate the LEF21 and beta2catenin signaling pathways[j ]. Proc Natl Acad Sci USA,1998,95 (8) :437424379 [13 ] Delcommenne M, Tan C, Gray V. et al. Phosphoinositide232OH kinase2dependent regulation of glycogen synthase kinase 3 and protein kinase BΠAKT by the integrin2linked kinase[j ]. Proc Natl Acad Sci USA,1998,95(19) :11211211216 [14 ] Hehlgans S, Haase M, Cordes N, et al. Signalling via integrins : Implications for cell survival and anticancer strategies [J ]. Biochim Biophys Acta,2007,1775(1) :1632180 [15 ] Nakrieko KA, Vespa A, Mason D, et al. Modulation of integrin2 linked kinase nucleo2 cytoplasmic shuttling by ILKAP and CRM1[J ]. Cell Cycle,2008,7(14) :215722166 [16 ] Kumar AS, Naruszewicz I, Wang P, et al. ILKAP regulates ILK signaling and inhibits anchorage2 independent growth[j ]. Oncogene, 2004,23 (19) :345423461 [17 ] Huang Z, Yan DP, Ge BX. JNK regulates cell migration through promotion of tyrosine phosphorylation of paxillin [ J ]. Cell Signal, 2008,20 (11) :200222012 [18 ] Sriraman V, Modi SR, Bodenburg Y, et al. Identification of ERK and JNK as signaling mediators on protein kinase C activation in cultured granulosa cells[j ]. Mol Cell Endocrinol, 2008, 294 (122) : 52260 [19 ] Sury MD, Agarinis C, Widmer HR. JNK is activated but does not mediate hippocampal neuronal apoptosis in experimental neonatal pneumococcal meningits[j ]. Neurobiol Dis, 2008,32(1) :1422150 [20 ] Uchida C, Gee E, Ispanovic E, et al. JNK as a positive regulator of angiogenic potential in endothelial cells[j ]. Cell Biol Int, 2008,32 (7) :7692776 [21 ] Lu G, Wang Y. Functional diversity of mammalian type 2C protein

494 25 phosphatase isoforms : new tales from an old family [J ]. Clin Exp Pharmacol Physiol, 2008,35(2) :1072112 [22 ] Kim AH, Khursigara G, Sun X, et al. Akt phosphorylates and negatively regulates apoptosis signal2regulating kinase 1[J ]. Mol Cell Biol,2001,21 (3) :8932901 [23 ] Tamura S, Toriumi S, Saito J, et al. PP2C family members play key roles in regulation of cell survival and apoptosis [ J ]. Cancer Sci, 2006,97 (7) :5632567 [24 ] Cengiz B, Gunduz M, Nagatsuka H, et al. Fine deletion mapping of chromosome 2q21237 shows three preferentially deleted regions in oral cancer[j ]. Oral Oncol, 2007,43 (3) :2412247 5 The Fifth Editorial Board of Chinese Journal of Biochemistry and Molecular Biology (Advisors) ZHENGJi ( Editor2in2Chief) J IA Hong2Ti (Associate Editors2in2Chief) CHANG Zeng2Yi LI Lin SHANG Yong2Feng SUN Zhi2Xian WANGLin2Fang WANG Zhi2Zhen YANG Fu2Yu ZHA Xi2Liang (Members of the Board,alphabetically) CHANG Zeng2Yi CHEN Qing2Xi GENG Yun2Qi GU Jun HANG Hai2Ying HE Rong2Qiao HUANGLi J IA Hong2Ti J IANG Cheng2Yu J IAO Bing2Hua KE Yang J IN You2Xin LI Bo2Liang LI Gang LI Gen2Xi LI Gui2Yuan LI Lin LI Ning LI Zai2Ping LIANG Ai2Hua LIANG Song2Ping LIN Qi2Shui LIU De2Fu LIU De2Pei LIU Guo2Qin LIU Jin2yuan MIAO Shi2Ying PENGJing2Pian QIAN Guan2Xiang QIANG Bo2Qin QU Liang2Hu RAO Zi2He SHI Yun2Yu RUAN Kang2Cheng SHANG Yong2Feng SHOU Cheng2Chao SUN Zhi2Xian WANGJia2Xi WANGLin2Fang WANG Zhi2Zhen WEI Qun WEN Jin2Kun XU Gen2Jun YANG Fu2Yu YANG Ke2Gong YANG Xiao2Ming YAO Li2Bo YAO Ren2Jie YE Qi2Nong YUAN Qin2Sheng ZHA Xi2Liang ZHANGJin ZHANG Nai2Heng ZHANG Xu2Jia ZHANG Yi ZHOU Chun2Yan ZHOU Hai2Meng ZHOU Jun2Mei ZHU Da2Hai ZHU Wei2Guo ZHU Yu2Xian (Specially Invited Members of the Board) Ray WU(USA) Robert K. YU(USA)