[1]柯东平,毛俊倩,刘 琪,等.TRIM21调控人结直肠癌细胞增殖分子机制研究*[J].陕西医学杂志,2019,(6):691-695.
　KE Dongping,MAO Junqian,LIU Qi,et al.Study on molecular mechanisms of TRIM21 in regulating proliferation of colorectal cancer[J].,2019,(6):691-695.
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TRIM21调控人结直肠癌细胞增殖分子机制研究*
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2019年6期
页码:
691-695
栏目:
基础研究
出版日期:
2019-06-05

文章信息/Info

Title:
Study on molecular mechanisms of TRIM21 in regulating proliferation of colorectal cancer
文章编号:
DOI:〖HT5K〗10.3969/j.issn.1000-7377.2019.06.003
作者:
柯东平1; 毛俊倩1; 刘 琪2; 郭甲民1; 马龙安
1.陕西省肿瘤医院普外科 （西安710061）；2.陕西省杨凌示范区医院普外科（杨凌712100）
Author(s):
KE Dongping;  MAO Junqian;  LIU Qi; et al.
Shaanxi Provincial Cancer Hospital Department of General Surgery(Xi’an 710061)
关键词:
结直肠癌; TRIM21; 转化生长因子-β1; 增殖; Smad
Keywords:
Key words Colorectal cancer;  TRIM21; Transforming growth factor-β1;  Proliferation; Smad
分类号:
R392.3
文献标志码:
A
摘要:
摘 要 〖HT5K〗目的：探讨TRIM21通过正调控转化生长因子-β1（TGF-β1）-Smad2/3信号通路影响人结直肠癌细胞增殖的分子机制。方法：在人结直肠癌细胞HCT116中转染TRIM21 siRNA沉默TRIM21的表达，采用荧光定量PCR和Western blot检测转染效果；采用MTT法检测转染后24 h、48 h、72 h和96 h时的细胞增殖情况，Western blot检测增殖相关蛋白CyclinD1的表达；采用Annexin V/PI染色结合流式细胞术检测细胞凋亡情况；荧光定量PCR检测TGF-β1、Smad2和Smad3在mRNA水平上的表达，Western blot检测TGF-β1、Smad2/3在蛋白水平上的表达及Smad2/3的磷酸化，验证TRIM21 siRNA转染后对TGF-β1—Smad2/3通路的影响。结果：TRIM21 siRNA转染组HCT116细胞中的TRIM21表达量明显下降（P<0.01），转染后24 h、48 h、72 h和96 h的细胞活力显著高于阴性对照组（P<0.05），Cyclin D1表达量上调（P<0.01）。TRIM21 siRNA转染后的凋亡细胞比例显著下降（P<0.01）。TRIM21 siRNA转染组TGF-β1的表达降低（P<0.01），Smad2和Smad3的表达量基本不变（P>0.05），但Smad2/3的磷酸化水平显著下降（P<0.01）。结论：干扰TRIM21的表达促进人结直肠癌细胞HCT116细胞增殖、抑制凋亡，其作用机制可能与影响TGF-β1—Smad2/3信号通路的激活和CyclinD1表达有关。
Abstract:
Abstract Objective: To investigate the molecular mechanism of TRIM21 affecting the proliferation of human colorectal cancer cells by positively regulating transforming growth factor-β1(TGF-β1)-Smad2/3 signaling pathway. Methods:TRIM21 siRNA was transfected into human colorectal cancer cells HCT116 to down-regulate the expression of TRIM21, and the effect of transfection was detected by real-time PCR and western blot. MTT assay was used to detect cell proliferation at 24 h, 48 h, 72 h and 96 h after transfection and the expression of proliferation-related protein CyclinD1 was detected by western blot. The expression of TGF-β1, Smad2 and Smad3 mRNA was detected by real-time PCR.Western blot was used to detect the expression of TGF-β1 and Smad2/3 at the protein level and phosphorylation of Smad2/3. All of these were to demonstrate the effect of TRIM21 siRNA transfection on the TGF-β1-Smad2/3 pathway. Results: The expression of TRIM21 in HCT116 cells was significantly decreased in TRIM21 siRNA transfection group (P<0.01), and the cell viability at 24 h, 48 h, 72 h and 96 h after transfection was significantly higher than that in the negative control group (P<0.05). The expression of Cyclin D1 was also up-regulated after transfection (P<0.01). The results of flow cytometry showed that the proportion of apoptotic cells after TRIM21 siRNA transfection was significantly decreased (P<0.01). The results of real-time PCR and western blot showed that the expression of TGF-β1 in TRIM21 siRNA transfection group was significantly lower than that in the negative control group (P<0.01). The expression levels of Smad2 and Smad3 were not different among the groups (P>0.05), but the phosphorylation level of Smad2/3 was significantly decreased after TRIM21 siRNA transfection (P<0.01). Conclusion: Down-regulation of TRIM21 can promote proliferation and inhibit apoptosis in human colorectal cancer cells HCT116, and its mechanism may be related to affecting the activation of TGF-β1-Smad2/3 signaling pathway and the expression of CyclinD1.

参考文献/References:

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备注/Memo

备注/Memo:
*陕西省重点科技创新项目(2014KCT-24)
更新日期/Last Update: 2019-06-20