Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (50): 9356-9359.doi: 10.3969/j.issn.1673-8225.2011.50.012

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Combined effects of pulse electric fields and wild-type p53 gene on apoptosis of Hela cells

Li Hao-shan1, 2, Xiong Zheng-ai1, Zhou Wei1, Li Cheng-xiang3, Yao Chen-guo3, Sun Cai-xin3   

  1. 1Department of Gynecology & Obstetrics, Second Affiliated Hospital of Chongqing Medical University, Chongqing  400010, China
    2Department of Gynecology & Obstetrics, Huaihe Hospital of Henan University, Kaifeng  475000, Henan Province, China
    3State Key Laboratory of Power Transmission Equipment & System Security and New Technology, Chongqing University, Chongqing  400010, China
  • Received:2011-04-11 Revised:2011-06-02 Online:2011-12-10 Published:2011-12-10
  • Contact: Xiong Zheng-ai, M.D., Department of Gynecology & Obstetrics, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China xiongzhengai61@21cn.com
  • About author:Li Hao-shan★, Master, Department of Gynecology & Obstetrics, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China; Department of Gynecology & Obstetrics, Huaihe Hospital of Henan University, Kaifeng 475000, Henan Province, China dashan0328@126.com
  • Supported by:

    Key Program of the National Natural Science Foundation of China, No. 50637020*; the Natural Science Foundation of Chongqing, No. CSTC2009BA5039*

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Abstract:

BACKGROUND: There have been lots of experimental works of animals and cells on pulses electric fields treating tumors. But electric fields combined with gene transfer had been seldom studied. Wild-type p53 gene combined with pulses electric fields (PEFs) can induce apoptosis of carcinoma cells.
OBJECTIVE: To investigate apoptosis of Hela cells induced by pulse electric fields combined with wild-type p53 gene and the mutual effect between them.
METHODS: PEFs with 100 μs duration, 1 Hz frequency, 8 pieces pulses and 1 500 V/cm electric intensity were performed on Hela cells, Hela-vector and Hela-p53 cells. p53 mRNA expression was detected by RT-PCR. The inhibitory effect on Hela cells growth was detected by MTT assay. Hela cells apoptosis was detected by flow cytometry at 12 hours after PEFs were performed. The expression levels of Bax and Bcl-2 were detected by western blot analysis. Expression of casepase-3 in cells was stained with immunofluorescence and detected by laser scanning confocal microscopy.
RESULTS AND CONCLUSION: Apoptotic rate of Hela, Hela-vector and Hela-p53 were (6.48±2.06)%, (7.22±2.25)% and (17.41±2.62)%, respectively. The apoptotic rate was significantly higher in the wild-type p53 gene combined with PEFs group than in the other two groups. At the same time, p53 and caspase-3 mRNA expression was significantly higher in the wild-type p53 gene combined with PEFs group than in the other groups. Bax/Bcl-2 expression ratio was significantly higher in the wild-type p53 gene combined with PEFs group than in the Hela cell group and Hela-vector group as detected by western blot analysis. Hela cells were inhibited by wild-type p53 gene transfer and wild-type p53 gene could increase Hela cells apoptosis induced by PEFs.

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