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[1]孙昊彤,马 璐,赵晓鹏,等.REV 7基因对人结肠癌细胞周期及凋亡的影响[J].宁夏医科大学学报,2018,(04):373-377.[doi:10.16050/j.cnki.issn1674-6309.2018.04.001]
 SUN Haotong,MA Lu,ZHAO Xiaopeng,et al.Effect of REV 7 Gene on Cell Cycle and Apoptosis of Human Colon Cancer Cells[J].Ningxia Medical University,2018,(04):373-377.[doi:10.16050/j.cnki.issn1674-6309.2018.04.001]
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REV 7基因对人结肠癌细胞周期及凋亡的影响(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2018年04期
页码:
373-377
栏目:
论 著
出版日期:
2019-09-30

文章信息/Info

Title:
Effect of REV 7 Gene on Cell Cycle and Apoptosis of Human Colon Cancer Cells
作者:
孙昊彤12 马 璐12 赵晓鹏12 李 昕12 隋 御12 徐 方12
(1.宁夏医科大学基础医学院,银川 750004; 2. 宁夏医科大学生育力保持教育部重点实验室,银川 750004)
Author(s):
SUN Haotong12 MA Lu12 ZHAO Xiaopeng12 LI Xin12 SUI Yu12 XU Fang12
(1. Ningxia Medical University,School of Basic Medical Sciences, Yinchuan 750004; 2. Ningxia Medical University Fertility Maintain Key Laboratory of Ministry of Education, Yinchuan 750004)
关键词:
结肠癌REV7基因细胞周期细胞凋亡DNA损伤修复
Keywords:
colon cancer REV7 gene cell cyclecell apoptosis DNA repair
分类号:
R735.3+5
DOI:
10.16050/j.cnki.issn1674-6309.2018.04.001
文献标志码:
A
摘要:
目的 探讨下调REV7基因对人结肠癌细胞HCT116、SW620细胞周期及凋亡的影响。方法 通过体外培养人结肠癌细胞株HCT116、SW620,用REV7基因的特异性siRNA片段转染细胞,运用实时荧光定量PCR、蛋白免疫印迹技术检测转染后细胞系的REV7基因干扰效率。选择REV7低表达具有统计学意义的上述细胞系作为实验组细胞,同时将加入转染试剂的HCT116、SW620细胞作为空白对照组,转染阴性RNA oligo干扰片段的HCT116、SW620细胞作为阴性对照组。采用流式细胞术测定各组细胞的细胞凋亡情况。通过Western blot法检测各组细胞抑癌蛋白P53、细胞周期蛋白P21、细胞周期蛋白依赖性激酶CDK4、细胞凋亡蛋白BAK、BCL-XL表达水平。结果 实时荧光定量PCR结果显示,HCT116、SW620细胞中siREV7干扰效率均高于60%(P<0.01);Annexin V-FITC/PI双染检测细胞凋亡结果显示,实验组与阴性对照相比,REV7下调促进HCT116、SW620细胞凋亡(P<0.05);Western blot结果显示,低表达REV7可下调HCT116、SW620细胞抗凋亡蛋白BCL-XL、细胞周期蛋白P21蛋白表达水平(P<0.01),上调促凋亡蛋白BAK、肿瘤抑制蛋白P53、细胞周期蛋白依赖性激酶CDK4蛋白表达水平(P<0.01)。结论 下调REV7可以阻滞结肠癌细胞HCT116、SW620细胞周期并促进细胞凋亡,其机制可能与下调REV7后,引起抑癌蛋白P53、蛋白细胞周期蛋白CDK4、P21及细胞凋亡蛋白的BAK、BCL-XL的表达变化有关。
Abstract:
Objective To investigate the effect of down-regulating REV7 gene on the cell cycle and apoptosis of human colon cancer cell lines HCT116 and SW620. Methods Specific siRNA fragment of REV7 gene was transfected into human colon cancer cell lines HCT116 and SW620 respectively. Real-time PCR,Western blot were used to detect the interference efficiency of REV7 in the transfected cell lines. Cells with low expression of REV7 was selected as the experimental group. Only transfection reagent added cells were used as blank control group. Transfected with negative RNA oligo interference fragments cells as negative control group. Flow cytometry(FCM) was used to determine the cell apoptosis in each group. The expression of tumor suppressor protein P53,cyclin protein P21 and CDK4,apoptosis protein BAK and BCL-XL were detected by Western blot. Results Real-time PCR results showed that the siREV7 interference efficiency in HCT116 and SW620 cells was higher than 60%(P<0.01). Annexin V-FITC/PI double staining showed that down-regulating REV7 promoted the early apoptosis of HCT116 and SW620 cells compared with the negative control in the experimental group(P<0.05). Western blot results showed that down-regulating REV7 decreased the expression of HCT116,BCL-XL and P21 in SW620 cells(P<0.01),increased the expression of pro-apoptotic protein BAK,tumor suppressor protein P53 and cyclin dependent kinase CDK4(P<0.01). Conclusion Down-regulation of REV7 can block the cell cycle of colon cancer cells HCT116 and SW620 and promote apoptosis. The mechanism may be related to the regulation of P53 protein by REV7 and the expression of cyclin CDK4,P21 and BAK and BCL-XL.

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备注/Memo

备注/Memo:
收稿日期:2017-12-30
基金项目:国家自然科学基金(31360251);国际交流合作项目(宁科技字[2016]25号);宁夏医科大学生殖与遗传优势学科群开放课题(XY201607)
作者简介:孙昊彤,女,在读硕士研究生,主要从事DNA损伤修复与肿瘤的发生发展的研究。
通信作者:徐方,女,博士研究生导师,主要从事DNA损伤修复与肿瘤的发生发展的研究。E-mail:xufang@nxmu.edu.cn
更新日期/Last Update: 2018-04-30