Please wait a minute...
Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2009, Vol. 3 Issue (4) : 384-389    https://doi.org/10.1007/s11684-009-0070-1
Research articles
The role of CDK1 siRNA interference in cell cycle and cell apoptosis
Hui XIAO PhD,Ming TIAN MM,Junna GE MM,Xin Wei MD,Zhaoming LI MM,Xiaolan LI MS,Deding TAO PhD,Junbo HU MD,Jianping GONG MD,
Cancer Research Institute, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;
 Download: PDF(212 KB)  
 Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract In the present report, cyclin-dependent kinase1 (CDK1) siRNA was transfected into cells to silence the CDK1 gene expression and study its role in the cell cycle and cell apoptosis. The siRNA targeting CDK1 gene was chemically synthesized and transfected into Hela cells by lipofectamine 2000. The expression levels of CDK1 gene and protein were examined by real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. The cell cycle was analyzed by using DNA content analysis by flow cytometry. Cell apoptosis was detected by the Annexin V/PI method. The morphological changes of transfected cells were examined under the microscopy by Wright-Giemsa stain. CDK1 gene was successfully silenced by its siRNA, and the CDK1 protein expression level was decreased significantly, especially from 48thh to 60thh after transfection. The DNA content analysis showed that transfection of CDK1 siRNA led to cells accumulating in G2/M phase. There was no significant difference in the apoptotic rate between transfected cells and the control cells after transfection of CDK1 siRNA for 48 or 60h. More double nucleus or multinucleus cells could be seen under the microscopy among the transfected cells. The decreased CDK1 expression by siRNA silencing gave rise to cell cycle arrest in G2/M phase but did not induce apoptosis.
Keywords cyclin-dependent kinase1      siRNA interference      cell cycle      apoptosis      
Issue Date: 05 December 2009
 Cite this article:   
Hui XIAO PhD,Ming TIAN MM,Junna GE MM, et al. The role of CDK1 siRNA interference in cell cycle and cell apoptosis[J]. Front. Med., 2009, 3(4): 384-389.
 URL:  
https://academic.hep.com.cn/fmd/EN/10.1007/s11684-009-0070-1
https://academic.hep.com.cn/fmd/EN/Y2009/V3/I4/384
Evan G I, Vousden K H. Proliferation, cell cycleand apoptosis in cancer. Nature, 2001, 411(6835): 342―349

doi: 10.1038/35077213
Blagosklonny M V. Apoptosis, proliferation, differentiation: in search of the order. Semin Cancer Biol, 2003, 13(2): 97―105

doi: 10.1016/S1044-579X(02)00127-X
Park M T, Lee S J. Cell cycle and cancer. J Biochem Mol Biol, 2003, 36(1): 60―65
Karin M, Lin A. NF-κB at the crossroadsof life and death. Nat Immunol, 2002, 3(3): 221―227

doi: 10.1038/ni0302-221
Vermeulen K, Berneman Z N, Van Bockstaele D R. Cell cycle and apoptosis. Cell Prolif, 2003, 36(3): 165―175

doi: 10.1046/j.1365-2184.2003.00267.x
Maddika S, Ande S R, Panigrahi S, Paranjothy T, Weglarczyk K, Zuse A, Eshraghi M, Manda K D, Wiechec E, Los M. Cell survival,cell death and cell cycle pathways are interconnected: implicationsfor cancer therapy. Drug Resist Updat, 2007, 10(1,2): 13―29
Morgam D O. Principles of CDK regulation. Nature, 1995, 374(6518): 131―134

doi: 10.1038/374131a0
Nurse P. Universalcontrol mechanism regulating onset of M phase. Nature, 1990, 344(6266): 503―508

doi: 10.1038/344503a0
Hunter T. Brakingthe cycle. Cell, 1993, 75(5): 839―841

doi: 10.1016/0092-8674(93)90528-X
Harlow E, Lane D. Antibodies: A LaboratoryManual. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1988, 450
Valcourt U, Gouttenoire J, Moustakas A, Herbage D, Mallein-Gerin F. Functions of transforminggrowth factor-beta family type I receptors and Smad proteins in thehypertrophic maturation and osteoblastic differentiation of chondrocytes. J Biol Chem, 2002, 277(37): 33545―33558

doi: 10.1074/jbc.M202086200
Kirou K A, Lee C, George S, Louca K, Papagiannis I G, Peterson M G, Ly N, Woodward R N, Fry K E, Lau A Y, Prentice J G, Wohlgemuth J G, Crow M K. Coordinateoverexpression of interferon-alpha-induce- genes in systemic lupuserythematosus. Arthritis Rheum, 2004, 50(12): 3958―3967

doi: 10.1002/art.20798
Kirou K A, Lee C, Crow M K. Measurement of cytokines in autoimmune disease. Methods Mol Med, 2004, 102: 129―154
Koopman G, Reutelingsperger C P, Kuijten G A, Keehnen R M, Pals S T, van Oers M H. Annexin V for flow cytometricdetection of phosphatidylserine expression of B cells undergoing apoptosis. Blood, 1994, 84(5): 1415―1420
Fadok V A, Voelker D R, Cammpbell P A, Cohen J J, Bratton D L, Henson P M. Exposure of phosphatidylserine on the surface of apoptoticlymphocytes triggers specific recognition and removal by macrophages. J Immunol, 1992, 148(7): 2207―2216
Bühler M, Moazed D. Transcription and RNAi inheterochromatic gene silencing. Nat StructMol Biol, 2007, 14(11): 1041―1048

doi: 10.1038/nsmb1315
Scoazec J Y. The RNA revolution. Ann Pathol, 2006, 26(4): 2752―2780
Tachikawa K, Briggs S P. Targeting the human genome. Curr Opin Biotechnol, 2006, 17(6): 659―665

doi: 10.1016/j.copbio.2006.10.010
Ying S Y, Chang D C, Miller J D, Lin S L. The microRNA:overview of the RNA gene that modulates gene functions. Methods Mol Biol, 2006, 342: 1―18
Zender L, Kubicka S. SiRNA based strategies forinhibition of apoptotic pathways in vivo-analytical and therapeuticimplications. Apoptosis, 2004, 9(1): 51―54

doi: 10.1023/B:APPT.0000012121.52210.23
Roberts J M. Evolving ideas about cyclins. Cell, 1999, 98(2): 129―132

doi: 10.1016/S0092-8674(00)81007-7
Nasmyth K. Viewpoint:putting the cell cycle in order. Science, 1996, 274(5293): 1643―1645

doi: 10.1126/science.274.5293.1643
Shi L, Nishioka W K, Th'ng J, Bradbury E M, Litchfield D W, Greenbergl A H. Premature p34cdc2 activation required for apoptosis. Science, 1994, 263(5150): 1143―1145

doi: 10.1126/science.8108732
Hahntow I N, Schneller F, Oelsner M, Weick K, Ringshausen I, Fend F, Peschel C, Decker T. Cyclin-dependent kinase inhibitorroscovitine induces apoptosis in chronic lymphocytic leukemia cells. Leukemia, 2004, 18(4): 747―755

doi: 10.1038/sj.leu.2403295
Golsteyn R M. Cdk1 and Cdk2 complexes (cyclin dependent kinases) in apoptosis:a role beyond the cell cycle. Cancer Lett, 2005, 217(2): 129―138

doi: 10.1016/j.canlet.2004.08.005
Rideout H J, Wang Q, Park D S, Stefanis L. Cyclin-dependentkinase activity is required for apoptotic death but not inclusionformation in cortical neurons after proteasomal inhibition. J Neurosci, 2003, 23(4): 1237―1245
Gartel A L. The conflicting roles of the cdk inhibitor p21(CIP1/WAF1) in apoptosis. Leuk Res, 2005, 29(11): 1237―1238

doi: 10.1016/j.leukres.2005.04.023
Davido D J, Leib D A, Schaffer P A. The cyclin-dependent kinase inhibitor roscovitine inhibitsthe transactivating activity and alters the posttranslational modificationof herpes simplex virus type 1 ICP0. JVirol, 2002, 76(3): 1077―1088
Edamatsu H, Gau C L, Nemoto T, Tamanoi F. Cdkinhibitors, roscovitine and olomoucine, synergize with farnesyltransferaseinhibitor (FTI) to induce efficient apoptosis of human cancer celllines. Oncogene, 2000, 19(27): 3059―3068

doi: 10.1038/sj.onc.1203625
Alvi A J, Austen B, Weston V J, Fegan C, Maccallum D, Gianella-Borradori A, Lane D P, Hubank M, Powell J E, Wei W, Taylor A M, Moss P A, Stankovic T. A novel CDK inhibitor, CYC202(R-roscovitine), overcomes the defect in p53-dependent apoptosis inB-CLL by down-regulation of genes involved in transcription regulationand survival. Blood, 2005, 105(11): 4484―4491

doi: 10.1182/blood-2004-07-2713
[1] Qiuxia Han, Hanyu Zhu, Xiangmei Chen, Zhangsuo Liu. Non-genetic mechanisms of diabetic nephropathy[J]. Front. Med., 2017, 11(3): 319-332.
[2] Xiaoling Wang,Yun Tan,Yizhen Li,Jingming Li,Wen Jin,Kankan Wang. Repression of CDKN2C caused by PML/RARα binding promotes the proliferation and differentiation block in acute promyelocytic leukemia[J]. Front. Med., 2016, 10(4): 420-429.
[3] Xiaowei GONG MD, PhD, Xiaoyan MING MD, Xu WANG MM, Daan WANG MD, Peng DENG MM, Yong JIANG MD, PhD, Aihua LIU MD, PhD, . Effect of PRAK gene knockout on the proliferation of mouse embryonic fibroblasts[J]. Front. Med., 2009, 3(4): 379-383.
[4] Dixin LI MM , Hongbing ZENG MD , Chunyang JI MM , . Effect of pirfenidone on renal tubulointerstitial fibrosis[J]. Front. Med., 2009, 3(3): 316-322.
[5] Yuan ZHANG MD , Xiaoyan ZHANG MM , Yanhui LI MM , Xuan DU MM , Zehua WANG MD, PhD , Hongbo WANG MD , . Effects of phosphatidylinositol 3-kinase inhibitor on human cervical carcinoma cells [J]. Front. Med., 2009, 3(3): 341-346.
[6] Qiong CHEN, Qing YU, Yuhu SONG, Peiyuan Li, Ying CHANG, Zhijun WANG, Lifeng LIU, Wei WU, Jusheng LIN. Cloning of human XAF1 gene promoter and assay of its transcription activity in a variety of cell lines[J]. Front Med Chin, 2009, 3(2): 148-152.
[7] Jianwen REN, Zhenhui PENG, Birong GUO, Min PAN. Celecoxib in combination with retinoid CD437 inhibits melanoma A375 cell in vitro[J]. Front Med Chin, 2009, 3(1): 108-112.
[8] ZHANG Shilong, ZENG Fuqing, PENG Shibo, WANG Liang. Effect on proliferation and apoptosis of T24 cell lines via silencing DNMT1 with RNA interference[J]. Front. Med., 2008, 2(4): 374-379.
[9] ZHAN Rong, YU Qinghong, HUANG Haobo. Effect of arsenic trioxide on proliferation and apoptosis of U266 cells and its relationship with the expression variation of VEGF[J]. Front. Med., 2008, 2(4): 356-360.
[10] LU Yi, QU Bo, LIU Chang, YU Liang, Liu Xuemin, WANG Haohua, JIANG An, ZHANG Xiaogang. Mechanism of hepatocellular damage in rat caused by low serum selenium[J]. Front. Med., 2008, 2(3): 255-258.
[11] SU Yuan, JIN Yang, ZHANG Xiaoju, ZHOU Qiong, BAI Ming, ZHU Liping. Impact of siRNA targeting pirh2 on proliferation and cell cycle control of the lung adenocarcinoma cell line A549[J]. Front. Med., 2007, 1(4): 359-363.
[12] OUYANG Shan, ZHANG Qinghua, QIAO Fuyuan. Relationship between expression of hepatocyte grow factor and apoptosis of trophoblasts in hypertensive disorder complicating pregnancy[J]. Front. Med., 2007, 1(4): 386-389.
[13] CAO Jie, LI Wanglin, XIA Jie, TANG Weibiao, WANG Hui, CHEN Xiwen, XIAO Huanqing, LI Yuyuan, CHEN Xiaoping, DU Hong, CHEN Shanming. Absence of FHIT expression is associated with apoptosis inhibition in colorectal cancer[J]. Front. Med., 2007, 1(2): 147-156.
[14] TIAN Yongji, LI Guilin, GAO Jun, WANG Renzhi, KONG Yanguo, ZHANG Zhenxing, LI Shifang, TIAN Shiqiang, DOU Wanchen, ZHANG Bo. Construction of 6HRE-GFAP-Baxα system specific for glioma gene therapy[J]. Front. Med., 2007, 1(1): 49-53.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed