| 王士玉,王小云,靳成娟,徐圣杰,刘湘楠,邬素芳.萝卜硫素诱导RASD1的表达促进宫颈癌HELA细胞凋亡及周期阻滞的机制研究[J].,2019,19(20):3801-3806 |
| 萝卜硫素诱导RASD1的表达促进宫颈癌HELA细胞凋亡及周期阻滞的机制研究 |
| Inhibition of Sulforaphane on the Proliferation of Cervical Cancer HELA Cells Via Up-regulating RASD1 |
| 投稿时间：2019-03-28 修订日期：2019-04-23 |
| DOI：10.13241/j.cnki.pmb.2019.20.001 |
| 中文关键词: 萝卜硫素（SFN） RASD1 凋亡 细胞周期阻滞 |
| 英文关键词: Sulforaphane(SFN) RASD1 Apoptosis Cell cycle arrest |
| 基金项目:国家重点研发计划"宫颈癌筛查与干预新技术及方案的研究"项目(2016YFC1302900) |
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| 中文摘要: |
| 摘要 目的：探讨萝卜硫素靶向RASD1，对宫颈癌细胞(HELA)凋亡及周期的影响机制。方法：不同浓度的萝卜硫素（SFN）作用于HELA细胞48h，采用CCK-8法检测SFN对HELA细胞增殖的影响；采用RT-PCR法检测SFN对RASD1基因mRNA表达水平的影响；采用Western印迹法检测SFN对 RASD1蛋白表达水平的影响；采用流式细胞技术检测加药以及过表达RASD1对HELA细胞凋亡及周期的影响，采用Western blot检测HELA中 cleaved-caspase3和cleaved-parp相关凋亡蛋白以及p21和cdc2周期相关蛋白的影响。结果：SFN抑制宫颈癌细胞增殖，且加药组中RASD1的mRNA和蛋白表达水平明显高于对照组，差异有统计学意义（P<0.05）。流式细胞凋亡分析结果提示：加SFN或是过表达RASD1细胞组凋亡率明显高于对照组，Western blot结果表明加SFN或过表达RASD1后细胞cleaved-caspase3以及cleaved-parp蛋白水平升高，差异具有统计学意义（P<0.05）；ModFit LT软件分析结果表明加SFN或过表达RASD1细胞S期显著低于对照组，G2/M期细胞增多，同时Western blot结果示p21蛋白的表达水平增高，而cdc2蛋白表达降低，其差异有统计学意义（P<0.05）。结论：在宫颈癌HELA 细胞中RASD1作为SFN的作用靶点，诱导细胞凋亡和细胞周期阻滞。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of SFN targeting RASD1 on apoptosis and cell cycle of human cervical cancer HELA cells. Methods: HELA cells were treated with different concentrations of SFN for 48h. Cell proliferation was measured by CCK-8 assay. The mRNA and protein expression of RASD1 in HELA cells were detected by real time-PCR and Western blot, respectively. Then HELA cells were treated with SFN or not, transfected with RASD1 and Control vector for 48h, cells apoptosis and cycle were detected by Flow cytometry, Western blot detected the relative protein level. Results: SFN inhibited the proliferation of HELA cells in a concentration-time dependent manner. Furthermore, SFN treatment could induce the expression of RASD1 at the mRNA and protein levels. Flow cytometry results suggested that the apoptosis ratio of cells with SFN or over-expression RASD1 cells was significantly higher than their control group. Western blot results suggested that the expression levels of cleaved-caspase3 and cleaved-parp were up-regulated than control group (P<0.05). ModFit LT analysis demonstrated that in the SFN plus group or RASD1 over-expression group the percentage of cells in S phase less than control group, and more than in G2/M phase clearly. Western blot shows cells with SFN or over expressing RASD1 remarkable increasing the level of p21 protein and decreasing the level of cdc2 protein (P＜0.05). Conclusion: RASD1 as a target of SFN inducing HELA cells apoptosis and cell cycle arrest. |
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