[1]梁冰),王峰),刘景超),等.沉默CTHRC1表达对乳癌MDAMB157细胞体外侵袭转移能力的影响[J].郑州大学学报(医学版),2013,(04):493.
 LIANG Bing,WANG Feng,LIU Jingchao,et al.Effect of silencing CTHRC1 on invasion and metastasis ability of breast cancer cell line MDAMB157 in vitro[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2013,(04):493.
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沉默CTHRC1表达对乳癌MDAMB157细胞体外侵袭转移能力的影响
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2013年04期
页码:
493
栏目:
论著
出版日期:
2013-07-20

文章信息/Info

Title:
Effect of silencing CTHRC1 on invasion and metastasis ability of breast cancer cell line MDAMB157 in vitro
作者:
梁冰1)王峰2)刘景超1)陈建中1)#
1)郑州大学第二附属医院乳腺外科 郑州 4500142)郑州大学第一附属医院肿瘤科 郑州 450052
Author(s):
LIANG Bing1) WANG Feng2) LIU Jingchao1) CHEN Jianzhong1)
1)Department of Breast Surgery, the Second Affiliated Hospital,Zhengzhou University, Zhengzhou 450014 2)Department of Oncology, the First Affiliated Hospital,Zhengzhou University, Zhengzhou 450052
关键词:
胶原三股螺旋重复蛋白1乳癌RNA干扰侵袭转移
Keywords:
collagen triple helic repeat containing 1breast cancerRNA interferenceinvasionmetastasis
分类号:
R737.9
摘要:
目的:探讨沉默CTHRC1基因表达对乳癌MDAMB157细胞体外侵袭转移能力的影响。 方法:化学合成靶向CTHRC1基因的siRNA,分别以不同浓度(10、20和30 nmol/L)转染MDAMB157细胞,同时以转染阴性对照siRNA和未转染细胞为对照。转染48 h后,应用RTPCR和Western blot检测细胞CTHRC1 mRNA和蛋白的表达情况。以干扰效率最高的CTHRC1 siRNA浓度再次转染MDAMB157细胞,同时以转染阴性对照siRNA、转染Lipofectamine 2000和未转染细胞为对照,分别在转染前、转染24、48和72 h后收集细胞,Boyden Chamber检测细胞侵袭能力的变化;转染48 h后做细胞划痕实验,分别于划痕后24、48和72 h检测划痕宽度。结果:各组细胞CTHRC1 mRNA和蛋白表达的差异有统计学意义(F=28.185和17.776,P<0.001)。随着CTHRC1 siRNA转染浓度的增加,CTHRC1 mRNA和蛋白的表达下降,30 nmol/L CTHRC1 siRNA干扰效率最高。MDAMB157细胞转染30 nmol/L CTHRC1 siRNA后,穿膜细胞数和划痕愈合能力明显下降(P<0.05)。 结论:抑制CTHRC1基因的表达能有效抑制乳癌MDAMB157细胞体外的侵袭和转移能力。
Abstract:
Aim: To explore the effect of silencing CTHRC1 by RNAi technique on the invasion and metastasis ability of breast cancer cell line MDAMB157.Methods:Chemically synthesized siRNA targeting CTHRC1 gene was transfected into MDAMB157 cells at different concentrations (10, 20, 30 nmol/L) respectively, and MDAMB157 cells transfected with negative control siRNA and untransfected MDAMB157 cells were used as control. 48 h after transfection, RTPCR and Western blot were used to detect the expressions of CTHRC1 mRNA and protein. The CTHRC1 siRNA with the highest interference efficiency was transfected into MDAMB157 cells again, and MDAMB157 cells transfected with negative control siRNA, or Lipofectamine 2000 and untransfected cells were used as control. Boyden chamber was used to detect the invasion ability of MDAMB157 cells before transfection and 24, 48, 72 h after transfection. 48 h after transfection, wound healing assay was used to detect the migration ability of MDAMB157 cells.Results: The expressions of CTHRC1 mRNA and protein among the groups were obviously different (F=28.185 and 17.776,P<0.001);with the increase of concentration of CTHRC1 siRNA, the expressions of CTHRC1 mRNA and protein decreased; with the 30 nmol/L CTHRC1 siRNA transfection, the interference efficiency reached the highest. After transfected with 30 nmol/L CTHRC1 siRNA, the numbers of MDAMB157 cells which traversed the membrane and the ability of wound healing decreased greatly(P<0.05). Conclusion: The invasion and metastasis ability of MDAMB157 cells could be decreased efficiently after inhibiting the expression of CTHRC1.

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备注/Memo

备注/Memo:
#通讯作者,男,1959年7月生,本科,主任医师,研究方向:乳癌病因,Email:jzchen59@yahoo.com.cn
更新日期/Last Update: 2013-07-25