Effects of tripartite motif containing 25 gene silencing on cisplatin resistance and apoptosis in A549/DDP cells

Su Lin; Qin Xia; Jing Huaizhi; Qu Can; Qiu Feng

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Effects of tripartite motif containing 25 gene silencing on cisplatin resistance and apoptosis in A549/DDP cells

更新时间：2023-12-18

- Effects of tripartite motif containing 25 gene silencing on cisplatin resistance and apoptosis in A549/DDP cells
- Journal of Chongqing Medical University Vol. 38, Issue 11, Pages: 1607(2014)
- 作者机构：
  
  重庆医科大学第一附属医院药学部
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Published：2014，
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Su Lin, Qin Xia, Jing Huaizhi, et al. Effects of tripartite motif containing 25 gene silencing on cisplatin resistance and apoptosis in A549/DDP cells. [J]. Journal of Chongqing Medical University 38(11):1607(2014)
DOI：

Su Lin, Qin Xia, Jing Huaizhi, et al. Effects of tripartite motif containing 25 gene silencing on cisplatin resistance and apoptosis in A549/DDP cells. [J]. Journal of Chongqing Medical University 38(11):1607(2014) DOI：

摘要

目的：三联基序蛋白２５（tripartite motif containing 25，TRIM25）为泛素化E3连接酶，通过抑制TRIM25表达，研究其对肺癌顺铂（cis-dichlorodiamine platinum，DDP）耐药细胞株A549/DDP耐药性及凋亡的影响。方法：针对TRIM25基因设计4条不同序列shRNA，分别构建重组质粒载体pt1、pt2、pt3、pt4。利用脂质体将质粒转染至A549/DDP细胞，采用real-time PCR、Western blot检测抑制效果，MTT实验和流式细胞仪（flow cytometry，FCM）实验分别检测在DDP作用下细胞耐药性改变和凋亡情况。结果：干扰质粒pt1在mRNA和蛋白质水平上均能特异性地抑制TRIM25的表达；MTT法结果显示，抑制TRIM25表达后，干扰组pt1细胞IC50值低于pNC组（P=0.001）及对照组（P=0.000）；FCM结果显示，干扰组pt1的细胞凋亡率明显低于pNC组（P=0.005）、对照组（P=0.004）。结论：抑制 TRIM25表达后，能有效地降低 A549/DDP细胞株的DDP耐药性并促进其凋亡。

Abstract

Objective：To research the effects of silencing tripartite motif containing 25（TRIM25） gene on cisplatin resistance and apop－tosis in cisplatin resistant lung cancer cell lines A549/DDP. Methods：Four types of short hairpin RNA were designed and prepared. The recombinant plasmids（pt1，pt2，pt3，pt4） were constructed respectively，which were transfected into A549/DDP cells using lipofectamine. Real-time PCR and Western blot were used to detect the inhibitory effects. The resistance of cisplatin to A549/DDP cells was detected by MTT assay. Cell apoptosis was tested by flow eytometry（FCM）. Results：The recombinant plasmid pt1 can inhibit mRNA and protein expression of TRIM25 specifically. MTT data showed that the cisplatin IC50 was decreased significantly in pt1 group than in pNC group（P=0.000） and control group（P=0.000） after the TRIM25 gene inhibition. According to the results of FCM，apoptotic cells was significantly decreased in pt1 group than in pNC group（P=0.005） and control group（P=0.004）. Conclusion：TRIM25 gene silencing in A549/DDP cells can effectively reduce cisplatin resistance and promote cells apoptosis.

关键词

siRNATRIM25A549/DDP细胞肿瘤耐药

Keywords

siRNAtripartite motif containing 25A549/DDPcellstumor resistance

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