Effects of transient receptor potential V6 silence on proliferation and apoptosis of trophoblasts

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Author:
WENG Zhan-ping(Department of Obstetrics, Qingdao Municipal Hospital, Qingdao 266071, China)
WANG Xue-mei(Department of Obstetrics, Qingdao Municipal Hospital, Qingdao 266071, China)
QI Hong()
TAO Hong(Department of Obstetrics, Qingdao Municipal Hospital, Qingdao 266071, China)
ZHANG Shu-ping(Department of Obstetrics, Qingdao Municipal Hospital, Qingdao 266071, China)
JI Xiang-hong(Department of Obstetrics, Qingdao Municipal Hospital, Qingdao 266071, China)
Journal Title:
Chinese Journal of Obstetrics and Gynecology
Issue:
Volume 47, Issue 10, 2012
DOI:
10.3760/cma.j.issn.0529-567x.2012.10.012
Key Word:
Calcium channels; TRPV cation channels; RNA, small interfering; Trophoblasts;Cell proliferation ; Apoptosis

Abstract: Objective To investigate the effects of the transient receptor potential V6 (TRPV6) gene silencing on the proliferation and apoptosis of trophoblasts HTR-8/SVneo cells.Methods siRNA sequences targeting the TRPV6 gene were constructed and then transfected into HTR-8/SVneo cells mediated by liposome.The cells were divided three groups,including blank control (add the reagent of transfenction),negative control groups (transfecting nonspecific siRNA) and experimental groups (transfecting TRPV6-siRNA).Those cells in every group were collected at 24,48,72 hours after transfecting.The expression levels of TRPV6 mRNA were detected by reverse transcription (RT) PCR at different times after transfecting.The effects of siRNA on the proliferation and apoptosis of the cells were assayed by methyl thiagolyl tetragolium (MTT) and flow cytometry at different times after transfecting.Results siRNA TRPV6 transfection could inhibit the expression of TRPV6 mRNA in the HTR-8/SVneo cells.The expression was decreased with the extension of time,by 0.72 ± 0.02,0.54 ± 0.02 and 0.29 ± 0.01 after 12,48 and 72 hours of siRNA transfection as compared with the blank control and the negative control groups (P <0.01).The rates of proliferation inhibition were (19.29 ± 1.23) %,(32.12 ± 1.35) % and (46.51 ±1.42) % at 24,48 and 72 hours respectively when compared with the blank control (2.12 ± 0.03)%,(2.42 ± 0.02) %,(3.13 ± 0.04) % and the negative control groups (2.37 ± 0.01) %,(2.61 ± 0.05) %,(2.93 ± 0.03) % (P < 0.01).The apoptosis rates of HTR-8/SVneo cells was 16.21% at 48 hours after transfected with siRNA TRPV6,which were significantly higher than 3.27% in the blank control and 5.34% in the negative control groups (P < 0.05).Conclusion Silenceing of TRPV6 genen could inhibit the proliferation and increase the apoptosis of extravillous trophoblas of human placenta.

  • [1]Hoenderop JG,van der Kemp AW,Hartog A,et al.Molecular identification of the apical Ca2 + channel in 1,25-dihydroxyvitamin D3-responsive epithelia.J Biol Chem,1999,274:8375-8378.
  • [2]李福春,谷贵山,金成浩,等.TRPV5、TRPV6在骨肉瘤、骨巨细胞瘤的表达.山东医药,2007,47:73-74.
  • [3]Landowski CP,Bolanz KA,Suzuki Y,et al.Chemical inhibitors of the calcium entry channel TRPV6.Pharm Res,2011,28:322-330.
  • [4]Kovacs G,Danko T,Bergeron MJ,et al.Heavy metal cations permeate the TRPV6 epithelial cation channel.Cell Calcium,2011,49:43-55.
  • [5]Haché S,Takser L,LeBellego F,et al.Alteration of calcium homeostasis in primary preeclamptic syncytiotrophoblasts:effect on calcium exchange in placenta.J Cell Mol Med,2011,15:654-667.
  • [6]Cosens DJ,Manning A.Abnormal electroretinogram from a Drosophila mutant.Nature,1969,224:285-287.
  • [7]Ramsey IS,Delling M,Clapham DE.An introduction to TRP channels.Annu Rev Physiol,2006,68:619-647.
  • [8]Clapham DE,Julius D,Montell C,et al.International Union of Pharmacology.XLIX.Nomenclature and structure-function relationships of transient receptor potential channels.Pharmacol Rev,2005,57:427-450.
  • [9]Hellwig N,Albrecht N,Harteneck C,et al.Homo-and heteromeric assembly of TRPV channel subunits.J Cell Sci,2005,118:917-928.
  • [10]Peng JB,Chen XZ,Berger UV,et al.Human calcium transport protein CaT1.Bioehem Biophys Res Commun,2000,278:326-332.
  • [11]Wasserman RH.Comments on the essentiality of calbindin-D9k and calcium channel TRPV6 for optimal active intestinal calcium transport in vitro.Ann N Y Acad Sci,2010,1192:365-366.
  • [12]Moreau R,Daoud G,Bernatchez R,et al.Calcium uptake and calcium transporter expression by trophoblast cells from human term placenta.Biochim Biophys Acta,2002,1564:325-332.
  • [13]Suzuki Y,Kovacs CS,Takanaga H,et al.Calcium channel TRPV6 is involved in murine maternal-fetal calcium transport.J Bone Miner Res,2008,23:1249-1256.
  • [14]Qiu Q,Yang M,Tsang BK,et al.EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways.Reproduction,2004,128:355-363.
  • [15]Cunningham FG,Gant NF,Leveno KJ.威廉姆斯产科学.段涛,丰有吉,狄文,等,译.21版.济南:山东科学技术出版社,2006:659-660.
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