中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (50): 9356-9359.doi: 10.3969/j.issn.1673-8225.2011.50.012

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

脉冲电场联合野生型p53基因诱导人宫颈癌Hela细胞凋亡

李好山1, 2，熊正爱1，周  玮1，李成祥3，姚陈果3，孙才新3   

  1. 1重庆医科大学附属第二医院妇产科，重庆市  400010
    2河南大学淮河医院妇产科，河南省开封市475000
    3重庆大学输配电装备及系统安全与新技术国家重点实验室，重庆市 400030
  • 收稿日期:2011-04-11 修回日期:2011-06-02 出版日期:2011-12-10 发布日期:2011-12-10
  • 通讯作者: 熊正爱，医学博士，重庆医科大学附属第二医院妇产科，重庆市 400010 xiongzhengai61@21cn.com
  • 作者简介:李好山★，男，1983年生，河南省开封市人，汉族，2011年重庆医科大学毕业，硕士，主要从事妇科肿瘤研究。 dashan0328@126.com
  • 基金资助:

    国家自然科学基金重点项目(50637020)；重庆市自然科学基金重点项目(CSTC2009BA5039)。

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*

PDF

395

摘要：

背景：在前期研究基础上，设想将基因电转染作为脉冲电场治疗恶性肿瘤的辅助方法。
目的：探讨脉冲电场联合抑癌基因野生型p53基因诱导人宫颈癌hela细胞发生凋亡及相互作用。
方法：采用空质粒及含有野生型p53的质粒转染Hela细胞得到Hela-vector、Hela-p53细胞，将Hela细胞、Hela-vector和Hela-p53细胞分别施加固定脉宽100 μs、频率1 Hz、脉冲数8个、电场强度为1 500 V/cm的脉冲电场处理。
结果与结论：相同参数脉冲电场作用于各组细胞12 h后，MTT结果显示Hela-p53组细胞吸光度明显低于Hela组(P < 0.05)；流式细胞仪及RT-PCR检测结果显示Hela-p53组的早期凋亡率和p53 mRNA表达较Hela组明显增加；Western bolt及激光共聚焦显微镜结果显示与Hela组比较，Hela-p53组细胞Bax蛋白表达明显上升，而Bcl-2蛋白则明显下降，Casepase-3相对荧光强度明显增强。表明野生型p53基因对宫颈癌Hela细胞生长有明显的抑制作用，野生型p53基因可以提高脉冲电场诱导宫颈癌Hela细胞凋亡的作用。

关键词: 脉冲电场, Hela细胞, 野生型p53基因, 凋亡, 抑制

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.

中图分类号: