刘 佳, 郭 艳①, 孙开来, 富伟能. BAX 对Hep2 细胞系的凋亡及化疗敏感性的影响*[J]. 中国肿瘤临床, 2010, 37(8): 426-428. DOI: 10.3969/j.issn.1000-8179.2010.08.002
引用本文: 刘 佳, 郭 艳①, 孙开来, 富伟能. BAX 对Hep2 细胞系的凋亡及化疗敏感性的影响*[J]. 中国肿瘤临床, 2010, 37(8): 426-428. DOI: 10.3969/j.issn.1000-8179.2010.08.002
LIU Jia1, GUO Yan2, SUN Kailai1, FU Weineng1. Effect of BAX on Apoptosis of Hep-2 Cells and Their Sensitivity to 5-Aza-CdR[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(8): 426-428. DOI: 10.3969/j.issn.1000-8179.2010.08.002
Citation: LIU Jia1, GUO Yan2, SUN Kailai1, FU Weineng1. Effect of BAX on Apoptosis of Hep-2 Cells and Their Sensitivity to 5-Aza-CdR[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(8): 426-428. DOI: 10.3969/j.issn.1000-8179.2010.08.002

BAX 对Hep2 细胞系的凋亡及化疗敏感性的影响*

Effect of BAX on Apoptosis of Hep-2 Cells and Their Sensitivity to 5-Aza-CdR

  • 摘要: 目的:研究BAX 基因对人喉鳞状细胞癌Hep2 细胞系凋亡和对化疗药5- 氮杂-2'- 脱氧胞苷(5-Aza-CdR)敏感性的影响。方法:通过溴化二甲噻唑二苯四氨唑(MTT)比色法检测5-Aza-CdR对喉癌Hep2 细胞体外增殖的影响。采用脂质体转染方法将BAX 基因转入Hep2 细胞后,应用RT-PCR 法检测BAX 基因在Hep2 细胞中的表达。用流式细胞术检测Hep2 细胞凋亡以及细胞周期变化情况。结果:通过用不同浓度的5-Aza-CdR处理喉Hep2 细胞发现,5-Aza-CdR能抑制Hep2 细胞体外生长,且呈时间与剂量依赖性,发现体外增殖的半数抑制浓度(IC 50)为4 μ mol/L。通过流式细胞术检测发现5-Aza-CdR和BAX 均可以使Hep2细胞发生G1/S 期细胞阻滞,并且能促进细胞凋亡。5-Aza-CdR处理后,与对照组相比,Hep2 细胞G0/G1 期的细胞数量明显增加,由对照组的51.57% 增高到71.17% ,凋亡率也由对照组的1.67% 增加到13.96% 。转染BAX 后,RT-PCR 结果显示转染BAX 基因的喉鳞状细胞癌Hep2 细胞中BAX 表达显著增加(P<0.05),S 期Hep2 细胞百分比由空白对照组的33.29% 、转染空载体组的32.42% 分别减少至16.07% 和13.58% ,细胞凋亡率相对于空白对照组的1.67% 、转染空载体组2.04% ,转染BAX 基因组的7.13% 增加到23.74% ,具有显著性差异(P<0.05)。 本研究结果还表明同时转染BAX 基因及加药组的喉鳞状细胞癌Hep2 细胞凋亡率明显增加。结论:转染BAX 基因能诱导Hep2 细胞的凋亡,并能增加Hep2 细胞对药物5-Aza-CdR杀伤作用的敏感性。

     

    Abstract: Objective:To investigate the effect of BAX on the apoptosis of human laryngeal squamous cell carcinoma Hep 2 cells and their sensitivity to5-aza-2' deoxycitydine ( 5-Aza-CdR).Methods:The proliferation of Hep2 cells was detect-ed by methyl thiazolyl tetrazolium (MTT) colorimetry after treatment with 5-Aza CdR in vitro. The plasmid containing BAX cDNA was transfected into Hep2 cells. The mRNA level of BAX gene was detected by RT-PCR. Flow cytometry was used to analyze the apoptosis and cell cycle of Hep2 cells. Results: After treatment with different concentrations of 5-Aza-CdR, the proliferation of Hep 2 cells was inhibited by 5-Aza-CdR in vitro in a dose- and time-dependent manner, and the IC 50 was 4 μ mol /L. The resul ts of flow cytometry detection showed that both BAX and 5-Aza-CdR could block Hep 2 cells in G1/S phase and promote apoptosis. The percentage of cells in G0/G1 and the apoptotic rate of Hep 2 cells treated with 5-Aza-CdR were increased from51.57% to71.17% and from1.67% to13.96%, respectively. The RT-PCR results showed that transfec-tion of BAX significantly increased the expression of BAX gene level in Hep 2 cells ( P<0.05). After transfection, the percent-age of Hep 2 cells in S phase were decreased from 33.29% and 32.42% to16.07% and 13.58% in blank control group and empty vector transfected group, respectively. The apoptotic rate of Hep 2 cells transfected with plasmid containing BAX cD -NA was increased from7.13% to23.74%, with a significant difference from that of the blank control group and empty vector transfected group ( P<0.05). Conclusion:Transfection of BAX can induce apoptosis of Hep 2 cells and increase their sensitivi-ty to the killing effect of 5-Aza-CdR.

     

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