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

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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
Volume 47, Issue 10, 2012
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.

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