| 辛伐他汀通过调控Chk1基因的表达对宫颈癌细胞增殖、凋亡以及放疗敏感性的影响 |
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| 引用本文： | 万 莉,比丽克孜·艾克木,刘 娜. 辛伐他汀通过调控Chk1基因的表达对宫颈癌细胞增殖、凋亡以及放疗敏感性的影响[J]. 现代肿瘤医学, 2020, 0(18): 3138-3144. DOI: 10.3969/j.issn.1672-4992.2020.18.009 |
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| 作者姓名： | 万 莉 比丽克孜·艾克木 刘 娜 |
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| 作者单位： | 新疆医科大学附属中医医院妇产科，新疆 乌鲁木齐 830000 |
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| 基金项目： | 新疆维吾尔自治区自然科学基金（编号:2019D01C182） |
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| 摘 要： | 目的:探讨辛伐他汀（simvastatin,SVA）通过调控细胞周期检测点激酶1（checkpoint kinase 1,Chk1）基因表达对宫颈癌细胞增殖、凋亡及放疗敏感性的影响。方法:体外培养宫颈癌Hela细胞，采用不同浓度（3.25 μmol/L、6.25 μmol/L、12.5 μmol/L、25 μmol/L、50 μmol/L)SVA处理细胞，以0 μmol/L SVA为空白组（Con组）。采用MTT法检测SVA对细胞的增殖抑制率，筛选SVA最适浓度（SVA组）用于后续研究。流式细胞术检测各组细胞凋亡率。采用2、4、6、8 Gy剂量X射线照射细胞。将Chk1阴性对照质粒、Chk1 siRNA分别转染至Hela细胞，分别命名为si-NC、si-Chk1组；重组pcDNA 3.1/Chk1基因过表达质粒及空载体对照质粒分转染至SVA组细胞中，分别命名为SVA+pcDNA-Chk1组、SVA+pcDNA组。采用qRT-PCR与Western blot法检测各组细胞中Chk1 mRNA及蛋白表达水平；MTT法检测各组细胞增殖能力；流式细胞术检测细胞凋亡情况；克隆形成实验检测细胞放疗敏感性。结果:随着SVA浓度的增加Hela细胞增殖抑制率明显升高且呈剂量依赖性，SVA浓度为12.5 μmol/L时Hela细胞增殖抑制率约为50%，以此用作后续实验研究。SVA组Hela细胞凋亡率显著高于Con组，且不同剂量X射线照射后Hela细胞存活分数明显降低（P＜0.05）；SVA处理Hela细胞后可显著降低Chk1 mRNA及蛋白表达水平（P＜0.05）；si-Chk1组Hela细胞中Chk1蛋白表达水平及细胞存活分数均显著低于si-NC组（P＜0.05），细胞增殖抑制率及凋亡率均显著升高（P＜0.05）；SVA+pcDNA-Chk1组Hela细胞增殖抑制率及细胞凋亡率均显著低于SVA+pcDNA组（P＜0.05），细胞存活分数显著升高（P＜0.05）。结论:SVA可通过下调Chk1基因表达进而抑制宫颈癌细胞增殖并促使细胞凋亡，同时能够增强宫颈癌细胞放疗敏感性。
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| 关 键 词： | 辛伐他汀 细胞周期检测点激酶1 宫颈癌 细胞增殖 细胞凋亡 放疗敏感性 |
Effect of simvastatin on proliferation,apoptosis and radiosensitivity of cervical cancer cells by regulating the expression of Chk1 gene |
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| Wan Li,Bilikiz·Akmu,Liu Na. Effect of simvastatin on proliferation,apoptosis and radiosensitivity of cervical cancer cells by regulating the expression of Chk1 gene[J]. Journal of Modern Oncology, 2020, 0(18): 3138-3144. DOI: 10.3969/j.issn.1672-4992.2020.18.009 |
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| Authors: | Wan Li Bilikiz·Akmu Liu Na |
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| Affiliation: | Department of Obstetrics and Gynecology，the Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University，Xinjiang Urumqi 830000,China. |
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| Abstract: | Objective:To investigate the effect of simvastatin(SVA) on the proliferation,apoptosis and radiosensitivity of cervical cancer cells by regulating cell cycle checkpoint kinase 1(Chk1) gene expression.Methods:Hela cells were cultured in vitro,and cells were treated with different concentrations(3.25 μmol/L,6.25 μmol/L,12.5 μmol/L,25 μmol/L,50 μmol/L) of SVA,and 0 μmol/L SVA was used as blank group(Con group).MTT assay was used to detect the inhibition rate of SVA on cells,and the optimal concentration of SVA(SVA group) was screened for subsequent study.Flow cytometry was used to detect the apoptosis rate of each group.The cells were irradiated with X-rays at doses of 2,4,6,and 8 Gy.The Chk1 negative control plasmid and Chk1 siRNA were transfected into Hela cells,respectively,and named as si-NC and si-Chk1 groups.The recombinant pcDNA 3.1/Chk1 gene overexpression plasmid and the empty vector control plasmid were transfected into SVA group,and named as SVA+pcDNA-Chk1 group and SVA+pcDNA group,respectively.The expression levels of Chk1 mRNA and protein in each group were detected by qRT-PCR and Western blot.MTT assay was used to detect the proliferation of cells in each group.Flow cytometry was used to detect apoptosis,and colony formation assay was used to detect the sensitivity of cell radiotherapy.Results:With the increase of SVA concentration,the inhibition rate of Hela cell proliferation was significantly increased in a dose-dependent manner.The inhibition rate of Hela cell proliferation was about 50% when the concentration of SVA was 12.5 μmol/L,which was used as a follow-up experimental study.The apoptosis rate of Hela cells in SVA group was significantly higher than that in Con group,and the survival fraction of Hela cells was significantly decreased after X-ray irradiation(P＜0.05).SVA treatment significantly reduced the expression of Chk1 mRNA and protein after Hela cells(P＜0.05).The expression level of Chk1 protein and cell survival fraction in Hela cells of si-Chk1 group were significantly lower than those in si-NC group(P＜0.05),and the cell proliferation inhibition rate and apoptosis rate were significantly increased(P＜0.05).The proliferation inhibition rate and apoptosis rate of Hela cells in SVA+pcDNA-Chk1 group were significantly lower than those in SVA+pcDNA group(P＜0.05),and the cell survival fraction was significantly increased(P＜0.05).Conclusion:SVA can inhibit the proliferation of cervical cancer cells and promote apoptosis by down-regulating the expression of Chk1 gene,and can enhance the sensitivity of cervical cancer cells. |
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| Keywords: | simvastatin checkpoint kinase 1 cervical cancer proliferation apoptosis radiosensivity |
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