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中华诊断学电子杂志 ›› 2016, Vol. 04 ›› Issue (03) : 160 -163. doi: 10.3877/cma.j.issn.2095-655X.2016.03.005

所属专题: 文献

消化系统疾病与心身医学观

siRNA沉默细胞周期相关激酶基因对鼠结肠癌生长及其细胞凋亡的影响
孔令斌1, 贺军2, 张秋生2, 何恒2, 马翠梅2, 杜宗梅2, 庞道华3,()   
  1. 1. 272067 济宁医学院临床学院
    2. 272069 济宁医学院附属医院消化内科
    3. 济宁医学院公共卫生学院
  • 收稿日期:2016-02-29 出版日期:2016-08-26
  • 通信作者: 庞道华
  • 基金资助:
    山东省医药卫生科技发展计划(2013WS0335)

Effects of small interfering RNA silencing CCRK gene on the growth and apoptosis of colorectal carcinoma in nude mice

Lingbin Kong1, Jun He2, Qiusheng Zhang2, Heng He2, Cuimei Ma2, Zongmei Du2, Daohua Pang3,()   

  1. 1. Clinical College, 3Public Health College, Jining Medical University, Jining 272067, China
    2. Department of Gastroenterology, Affiliated Hospital of Jining Medical University, Jining 272069, China
  • Received:2016-02-29 Published:2016-08-26
  • Corresponding author: Daohua Pang
  • About author:
    Corresponding author: Pang Daohua, Email:
引用本文:

孔令斌, 贺军, 张秋生, 何恒, 马翠梅, 杜宗梅, 庞道华. siRNA沉默细胞周期相关激酶基因对鼠结肠癌生长及其细胞凋亡的影响[J/OL]. 中华诊断学电子杂志, 2016, 04(03): 160-163.

Lingbin Kong, Jun He, Qiusheng Zhang, Heng He, Cuimei Ma, Zongmei Du, Daohua Pang. Effects of small interfering RNA silencing CCRK gene on the growth and apoptosis of colorectal carcinoma in nude mice[J/OL]. Chinese Journal of Diagnostics(Electronic Edition), 2016, 04(03): 160-163.

目的

探讨siRNA沉默细胞周期相关激酶(CCRK)基因对鼠结肠癌生长及其凋亡的影响。

方法

将50只BALB/C(nu/nu)裸鼠皮下接种人类结肠癌SW480细胞,将其中建立移植瘤裸鼠模型成功的30只裸小鼠按照单纯随机抽样方法随机分为3组,分别接种siRNACCRK转染的人类结肠癌SW480细胞株(实验组)、正常培养的SW480细胞(空白对照组)和以转染携带无关序列片段sh RNA慢病毒的SW480细胞(阴性对照组)。第42天处死裸鼠后取下肿瘤组织,比较各组肿瘤重量、CCRKmRNA表达量、CCRK蛋白表达和细胞凋亡的变化。

结果

转染siRNA组肿瘤重量、CCRK mRNA和CCRK蛋白表达[(0.54±0.15)g,1.14±0.24和0.18±0.05]表达显著低于空白对照组[(1.05 ± 0.14)g,2.41±0.42和0.49±0.07]和空siRNA载体组[(1.07 ±0 .12)g,2.39±0.47,0.47±0.09;F=21.18,23.47,90.03;P<0.01];空白对照组与空siRNA载体组比较差异无统计学意义(q=0.98,1.07,1.13;P>0.05)。实验组沉默CCRK基因后组织中细胞凋亡率为(21.23±1.12)%,明显高于空白对照组(6.78±0.37)%和阴性对照组(7.25±0.32)%,差异有统计学意义(F=56.936,P<0.01);但空白对照组和空siRNA载体组之间差异无统计学意义(q=1.78,P>0.05)。

结论

CCRK靶向siRNA表达载体接种结肠癌裸鼠后能显著降低结肠癌瘤体的增长,抑制CCRK基因和蛋白表达;siRNA沉默CCRK基因能促进其凋亡,CCRK基因有望成为结直肠癌治疗的新靶点。

Objective

To investigate the effect of siRNA silencing cell cycle-related kinase (CCRK) gene on the growth and apoptosis of colon cancer in mice, and to provide a new idea and method for the treatment of colon cancer.

Methods

Fifty BALB / C (nu / nu) nude mice were inoculated with human colorectal carcinoma SW480 cells, in which 30 nude mice were successfully established for transplantation tumor model and were randomly divided into three groups, siRNA CCRK transfection of human SW480 colon cancer cell line (experimental group) and normal cultured SW480 cells (blank control group), and carrying irrelevant sequence fragment shRNA slow transfection of virus SW480 cells (negative control group) respectively.The nude mice were sacrificed and the tumor tissues were removed after 42 days.The tumor weight, CCRKmRNA expression, CCRK protein expression and apoptosis rate were compared between the groups.

Results

siRNA transfection group, tumor weight, CCRK mRNA and CCRK protein expression [(0.54±0.15)g, 1.14±0.24, 0.18±0.05] were significantly lower than those of the control group [(1.05±0.14)g, 2.41±0.42, 0.49±0.07] and empty vector of siRNA group [(1.07±0.12)g, 2.39±0.47, 0.47±0.09, F=21.18, 23.47, 90.03; P<0.01]. There were no significant differences between the blank control group and the empty siRNA vector group (q=0.98, 1.07, 1.13; P>0.05). Cells apoptosis rate in siRNA transfection group (21.23±1.12)% was significantly higher than that in normal control group (6.78±0.37)% and transfected with empty vector of siRNA group (7.25±0.32)% (P<0.01); but there was no significant difference between the blank control group and the empty siRNA vector group (q=1.78, P>0.05).

Conclusions

CCRK targeting siRNA expression vector inoculation with colon cancer can significantly reduce the growth of colon cancer and inhibit the expression of CCRK gene and protein in nude mice; CCRK gene silencing by siRNA can promote the colon cancer cells apoptosis; thus , CCRK gene can serve as a new potential molecular target for rectal cancer in gene therapy by siRNA.

图1 免疫组化法检测不同组别瘤组织CCRK蛋白表达(HE ×400)。示转染siRNA组(a)CCRK mRNA和CCRK蛋白表达明显低于空白对照组(b)和阴性对照组(c)(箭头所示);CCRK为细胞周期相关激酶
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