热汗古丽·库尔班,潘 静,吐松古·艾买尔,阿娜尔古丽,玛依拉,薄晓莉.eIF3A和eIF4E协同调控子宫颈癌细胞的增殖与凋亡[J].,2024,(14):2634-2638 |
eIF3A和eIF4E协同调控子宫颈癌细胞的增殖与凋亡 |
eIF3A and eIF4E Synergistically Regulate Proliferation and Apoptosis of Cervical Cancer Cells |
投稿时间:2023-11-29 修订日期:2023-12-26 |
DOI:10.13241/j.cnki.pmb.2024.14.006 |
中文关键词: 真核翻译起始因子 宫颈癌 增殖 凋亡 慢病毒 |
英文关键词: Eukaryotic translation initiation factor Cervical cancer Proliferation Apoptosis Lentivirus |
基金项目:省部共建中亚高发病成因与防治国家重点实验室开放课题项目(SKL-HIDCA-2022-GJ7) |
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中文摘要: |
摘要 目的:探讨eIF3A和eIF4E协同调控子宫颈癌细胞增殖与凋亡的相关机制。方法:选择对数生长期的人子宫颈癌细胞系HeLa、SiHa和正常宫颈上皮细胞HcerEpic,分别采用实时荧光定量PCR(qRT-PCR)和Western blot检测eIF 3A和eIF4E mRNA以及蛋白表达量。然后构建eIF 3A和eIF4E表达沉默的慢病毒载体,转染SiHa细胞后qRT-PCR和Western blot筛选稳定表达株。最后将SiHa细胞分为4组,即空白对照组、eIF 3A沉默组、eIF 4E沉默组和eIF 3A+eIF4E沉默组,连续培养48 h后CCK8实验检测细胞增殖率,流式细胞术检测细胞凋亡率,Western blot检测Ki-67和cleaved-Caspase-3蛋白表达量。结果:①与正常宫颈细胞相比,HeLa和SiHa中eIF 3A、eIF4E mRNA以及蛋白表达量均显著升高(P<0.05)。②与对照组相比,eIF 3A沉默组eIF 3A mRNA和蛋白表达量显著降低;eIF 4E沉默组eIF 4EmRNA和蛋白表达量显著降低;eIF 3A+eIF4E沉默组eIF 3A和eIF4E mRNA及蛋白表达量均显著降低(P<0.05)。③与对照组相比,eIF 3A沉默组、eIF 4E沉默组及eIF 3A+eIF4E沉默组细胞增殖率和Ki-67蛋白表达量下降,细胞凋亡率和cleaved-Caspase-3蛋白表达量升高,且eIF 3A+eIF4E沉默组细胞幅度最大(P<0.05)。结论:子宫颈癌中eIF 3A和eIF4E可能协同发挥调控细胞增殖与凋亡的生物学功能。 |
英文摘要: |
ABSTRACT Objective: To investigate the synergistic regulation of cervical cancer cell proliferation and apoptosis by eIF3A and eIF4E. Methods: Firstly, human cervical cancer cell lines HeLa, SiHa and normal cervical epithelial cell HcerEpic with logarithmic growth phase were selected. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot were used to detect eIF 3A, eIF4E mRNA and protein expression levels, respectively. Then lentiviral vectors for silent expression of eIF 3A and eIF4E were constructed and transfected SiHa cell, stable expression strains were screened using qRT-PCR and Western blot. Finally, SiHa cells were divided into four groups, namely the control group, eIF 3A low-expression group, eIF 4E low-expression group, and eIF 3A+eIF4E low-expression group. After continuous cultivation for 48 hours, cell proliferation rate was measured using CCK8 assay, cell apoptosis rate was measured using flow cytometry, and the expression levels of Ki-67 and cleared-Caspase-3 proteins were detected using Western blot. Results: ① Compared with normal cervical cells, the expression levels of eIF 3A, eIF4E mRNA and protein in HeLa and SiHa were significantly higher(P<0.05). ② Compared with the control group, the eIF 3A low-expression group significantly reduced the expression of eIF 3A mRNA and protein; the expression levels of eIF 4E mRNA and protein were significantly less in the eIF 4E low-expression group; the expression levels of eIF 3A, eIF4E mRNA and protein were both significantly lower in the eIF 3A+eIF4E low-expression group (P<0.05). ③Compared with the control group, the eIF 3A low-expression group, eIF 4E low-expression group and the eIF 3A+eIF4E low-expression group showed decrease of cell proliferation rate, increase of cell apoptosis rate, less of Ki-67 protein expression, and higher of cleared-Casase-3 protein expression (P<0.05). Conclusion: eIF 3A and eIF4E may synergistically exert biological functions in regulating cell proliferation and apoptosis of cervical cancer. |
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