沉默CDC20基因通过抑制Wnt/β-连环蛋白信号通路对子宫内膜癌细胞增殖和细胞周期的影响
R737.33; 目的:探讨细胞分裂周期蛋白20(CDC20)对子宫内膜癌(EC)细胞增殖和细胞周期的影响,并阐明其作用机制.方法:实时荧光定量PCR(RT-qPCR)和Western blotting法检测人子宫内膜基质 T-HESC 细胞和人 EC 细胞(KLE、RL95-2、ZJB-ENC1 和 ECC-1 细胞)中 CDC20 mRNA及蛋白表达水平,选择RL95-2细胞用于后续实验.将CDC20 shRNA干扰慢病毒转染至RL95-2细胞中,分为对照组、sh-NC组(感染阴性对照慢病毒)、sh-CDC20组(感染CDC20 shRNA干扰慢病毒)、sh-NC+SM04690组(感染阴...
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| Published in | 吉林大学学报（医学版） Vol. 50; no. 5; pp. 1305 - 1312 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | Chinese |
| Published |
齐齐哈尔医学院附属第三医院妇一科,黑龙江 齐齐哈尔 161000
28.09.2024
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1671-587X |
| DOI | 10.13481/j.1671-587X.20240514 |
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| Abstract | R737.33; 目的:探讨细胞分裂周期蛋白20(CDC20)对子宫内膜癌(EC)细胞增殖和细胞周期的影响,并阐明其作用机制.方法:实时荧光定量PCR(RT-qPCR)和Western blotting法检测人子宫内膜基质 T-HESC 细胞和人 EC 细胞(KLE、RL95-2、ZJB-ENC1 和 ECC-1 细胞)中 CDC20 mRNA及蛋白表达水平,选择RL95-2细胞用于后续实验.将CDC20 shRNA干扰慢病毒转染至RL95-2细胞中,分为对照组、sh-NC组(感染阴性对照慢病毒)、sh-CDC20组(感染CDC20 shRNA干扰慢病毒)、sh-NC+SM04690组(感染阴性对照慢病毒后,加入64 nmol·L-1 Wnt/β-连环蛋白信号通路抑制剂SM04690干预48 h)和sh-CDC20+SM04690组(感染CDC20 shRNA干扰慢病毒后,加入64 nmol·L-1 SM04690 干预 48 h).RT-qPCR法和 Western blotting法检测不同细胞中 CDC20 mRNA 和蛋白表达水平,CCK-8法检测各组细胞增殖活性,BrdU法检测各组细胞中BrdU阳性细胞百分率,流式细胞术检测各组G2/M期细胞百分率,Western blotting法检测各组细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平.结果:与T-HESC细胞比较,KLE、RL95-2、ZJB-ENC1和ECC-1细胞中CDC20 mRNA和蛋白表达水平均明显升高(P＜0.05),其中RL95-2细胞中CDC20 mRNA和蛋白表达水平最高.与sh-NC组比较,sh-CDC20组和sh-NC+SM04690组细胞增殖活性和BrdU阳性细胞百分率明显降低(P＜0.05),G2/M期细胞百分率明显升高(P＜0.05),细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平明显降低(P＜0.05).与sh-CDC20组比较,sh-CDC20+SM04690组细胞增殖活性和BrdU阳性细胞百分率明显降低(P＜0.05),G2/M期细胞百分率明显升高(P＜0.05),细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平明显降低(P＜0.05).结论:EC细胞中CDC20呈高表达,沉默CDC20可能通过调控Wnt/β-连环蛋白信号转导诱导RL95-2细胞G2/M期阻滞,抑制细胞增殖. |
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| AbstractList | R737.33; 目的:探讨细胞分裂周期蛋白20(CDC20)对子宫内膜癌(EC)细胞增殖和细胞周期的影响,并阐明其作用机制.方法:实时荧光定量PCR(RT-qPCR)和Western blotting法检测人子宫内膜基质 T-HESC 细胞和人 EC 细胞(KLE、RL95-2、ZJB-ENC1 和 ECC-1 细胞)中 CDC20 mRNA及蛋白表达水平,选择RL95-2细胞用于后续实验.将CDC20 shRNA干扰慢病毒转染至RL95-2细胞中,分为对照组、sh-NC组(感染阴性对照慢病毒)、sh-CDC20组(感染CDC20 shRNA干扰慢病毒)、sh-NC+SM04690组(感染阴性对照慢病毒后,加入64 nmol·L-1 Wnt/β-连环蛋白信号通路抑制剂SM04690干预48 h)和sh-CDC20+SM04690组(感染CDC20 shRNA干扰慢病毒后,加入64 nmol·L-1 SM04690 干预 48 h).RT-qPCR法和 Western blotting法检测不同细胞中 CDC20 mRNA 和蛋白表达水平,CCK-8法检测各组细胞增殖活性,BrdU法检测各组细胞中BrdU阳性细胞百分率,流式细胞术检测各组G2/M期细胞百分率,Western blotting法检测各组细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平.结果:与T-HESC细胞比较,KLE、RL95-2、ZJB-ENC1和ECC-1细胞中CDC20 mRNA和蛋白表达水平均明显升高(P＜0.05),其中RL95-2细胞中CDC20 mRNA和蛋白表达水平最高.与sh-NC组比较,sh-CDC20组和sh-NC+SM04690组细胞增殖活性和BrdU阳性细胞百分率明显降低(P＜0.05),G2/M期细胞百分率明显升高(P＜0.05),细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平明显降低(P＜0.05).与sh-CDC20组比较,sh-CDC20+SM04690组细胞增殖活性和BrdU阳性细胞百分率明显降低(P＜0.05),G2/M期细胞百分率明显升高(P＜0.05),细胞中β-连环蛋白、c-Myc和细胞周期素D1蛋白表达水平明显降低(P＜0.05).结论:EC细胞中CDC20呈高表达,沉默CDC20可能通过调控Wnt/β-连环蛋白信号转导诱导RL95-2细胞G2/M期阻滞,抑制细胞增殖. |
| Abstract_FL | Objective:To discuss the effect of cell division cycle protein 20(CDC20)on the proliferation and cell cycle of endometrial carcinoma(EC)cells,and to clarify its mechanism.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of CDC20 mRNA and protein in human endometrial stromal T-HESC cell and EC cells(KLE,RL95-2,ZJB-ENC1,and ECC-1 cells).The RL95-2 cells were selected for the subsequent experiments.CDC20 shRNA interference lentivirus was transfected into the RL95-2 cells and the cells were divided into control group,sh-NC group(infected with negative control lentivirus),sh-CDC20 group(infected with CDC20 shRNA interference lentivirus),sh-NC+SM04690 group(infected with negative control lentivirus followed by treatment with 64 nmol·L-1 Wnt/β-catenin signaling pathway inhibitor SM04690 for 48 h),and sh-CDC20+SM04690 group(infected with CDC20 shRNA interference lentivirus followed by treatment with 64 nmol·L-1 SM04690 for 48 h).RT-qPCR and Western blotting methods were used to detect the expression levels of CDC20 mRNA and proteins in the cells in various groups;CCK-8 method was used to detect the proliferation activities of the RL95-2 cells in various groups;BrdU assay was used to detect the percentages of BrdU positive cells in various groups;flow cytometry was used to detect the percentages of the cells at G2/M stage in various groups;Western blotting method was used to detect the expression levels of β-catenin,oncogene c-Myc,and cyclin D1 proteins in the cells in various groups.Results:Compared with T-HESC cells,the expression levels of CDC20 mRNA and protein in the KLE,RL95-2,ZJB-ENC1,and ECC-1 cells were significantly increased(P＜0.05),and the highest expression levels of CDC20 mRNA and protein were observed in RL95-2 cells.Compared with sh-NC group,the proliferation activities and percentages of the BrdU positive cells in sh-CDC20 group and sh-NC+SM04690 group were significantly decreased(P＜0.05),the percentages of the cells at G2/M phase were significantly increased(P＜0.05),and the expression levels of β-catenin,c-Myc,and cyclin D1 proteins were significantly decreased(P＜0.05).Compared with sh-CDC20 group,the proliferation activity and percentage of BrdU positive cells in sh-CDC20+SM04690 group were significantly decreased(P＜0.05),the percentage of the cells at G2/M phase was significantly increased(P＜0.05),and the expression levels of β-catenin,c-Myc,and cyclin D1 proteins in the cells were significantly decreased(P＜0.05).Conclusion:CDC20 is highly expressed in the EC cells.Silencing CDC20 may inhibit the cell proliferation by inducing G2/M phase arrest in the RL95-2 cells through the regulation of Wnt/β-catenin signal transduction. |
| Author | 赵薇 刘春静 杨钰杰 刘丽晶 王娜 |
| AuthorAffiliation | 齐齐哈尔医学院附属第三医院妇一科,黑龙江 齐齐哈尔 161000 |
| AuthorAffiliation_xml | – name: 齐齐哈尔医学院附属第三医院妇一科,黑龙江 齐齐哈尔 161000 |
| Author_FL | ZHAO Wei LIU Lijing LIU Chunjing WANG Na YANG Yujie |
| Author_FL_xml | – sequence: 1 fullname: LIU Chunjing – sequence: 2 fullname: YANG Yujie – sequence: 3 fullname: ZHAO Wei – sequence: 4 fullname: LIU Lijing – sequence: 5 fullname: WANG Na |
| Author_xml | – sequence: 1 fullname: 刘春静 – sequence: 2 fullname: 杨钰杰 – sequence: 3 fullname: 赵薇 – sequence: 4 fullname: 刘丽晶 – sequence: 5 fullname: 王娜 |
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| DocumentTitle_FL | Effect of silencing CDC20 on proliferation and cell cycle of endometrial cancer cells by inhibiting Wnt/β-catenin signaling pathway |
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| Keywords | 细胞周期 Cell proliferation Wnt/β-连环蛋白信号通路 Endometrial cancer Wnt/β-catenin signaling pathway 子宫内膜癌 Cell cycle 细胞分裂周期蛋白20 Cell division cycle protein 20 细胞增殖 |
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| PublicationTitle | 吉林大学学报（医学版） |
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| PublicationYear | 2024 |
| Publisher | 齐齐哈尔医学院附属第三医院妇一科,黑龙江 齐齐哈尔 161000 |
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| Snippet | R737.33; 目的:探讨细胞分裂周期蛋白20(CDC20)对子宫内膜癌(EC)细胞增殖和细胞周期的影响,并阐明其作用机制.方法:实时荧光定量PCR(RT-qPCR)和Western blotting法检测人子宫内膜基质 T-HESC 细胞和人 EC 细胞(KLE、RL95-2、ZJB-ENC1 和 ECC-1... |
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| Title | 沉默CDC20基因通过抑制Wnt/β-连环蛋白信号通路对子宫内膜癌细胞增殖和细胞周期的影响 |
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