CKS2 shRNA慢病毒表达载体的构建及其抑制卵巢癌A2780细胞增殖

被引：5

作者：

董梦梦

孙倩玲

张灵芝

张佳乐

袁成福

机构：

[1] 三峡大学医学院

来源：

基础医学与临床 | 2016年 / 36卷 / 02期

关键词：

CKS2; 卵巢癌; 细胞增殖; 选择性剪接;

D O I：

10.16352/j.issn.1001-6325.2016.02.001

中图分类号：

R737.31 [卵巢肿瘤];

学科分类号：

100214 [肿瘤学];

摘要：

目的构建靶向CKS2的shRNA慢病毒表达载体,并探讨CKS2敲除对卵巢癌细胞A2780增殖的影响。方法根据siRNA原理设计两对靶向CKS2的shRNA序列(shRNA1、shRNA2)及阴性对照NC shRNA,并克隆到慢病毒表达载体PLKO.1/puro中;将上述质粒转染到293T细胞并包装出慢病毒,用慢病毒感染卵巢癌细胞A2780,实时定量PCR及Western blot检测CKS2表达;MTT法检测A2780细胞增殖;Western blot检测PARP-1的剪切;流式细胞术检测凋亡。结果成功构建CKS2 shRNA慢病毒表达载体能明显降低A2780细胞中CKS2 mRNA及蛋白表达(P<0.05);能显著抑制A2780靶向CKS2的shRNA慢病毒表达载体,并包装出慢病毒;感染A2780细胞后,细胞增殖受抑(P<0.05);能诱导PARP-1蛋白的剪切及凋亡(P<0.05)。结论成功构建靶向CKS2的shRNA慢病毒表达载体,能显著抑制A2780细胞增殖。

引用

页码：145 / 150

页数：6

共 5 条

[1]

Expression and clinical significance of cyclin kinase subunit 2 in colorectal cancer [J].

Yu, Minhao ;

Zhong, Ming ;

Qiao, Zhongdong .

ONCOLOGY LETTERS, 2013, 6 (03) :777-780

[2]

Clinicopathological and biological significance of CDC28 protein kinase regulatory subunit 2 overexpression in human gastric cancer [J].

Tanaka, Fumiaki ;

Matsuzaki, Shinji ;

Mimori, Koshi ;

Kita, Yoshiaki ;

Inoue, Hiroshi ;

Mori, Masaki .

INTERNATIONAL JOURNAL OF ONCOLOGY, 2011, 39 (02) :361-372

[3]

Clinical significance and expression of cyclin kinase subunits 1 and 2 in hepatocellular carcinoma.[J].Dong‐YanShen;Zan‐XiFang;PanYou;Ping‐GuoLiu;FenWang;Chun‐LingHuang;Xiao‐BinYao;Zi‐XuanChen;Zhong‐YingZhang.Liver International.2009, 1

[4]

Clinical; pathological; proliferative and molecular responses associated with neoadjuvant aromatase inhibitor treatment in breast cancer.[J].W.R. Miller.Journal of Steroid Biochemistry and Molecular Biology.2009, 4

[5]

Microarray gene expression profiling in meningiomas: Differential expression according to grade or histopathological subtype [J].

Fevre-Montange, Michelle ;

Champier, Jacques ;

Durand, Anne ;

Wierinckx, Anne ;

Honnorat, Jerome ;

Guyotat, Jacques ;

Jouvet, Anne .

INTERNATIONAL JOURNAL OF ONCOLOGY, 2009, 35 (06) :1395-1407

← 1 →