人大肠癌lovo细胞株PARG基因沉默对蛋白激酶P38表达及其磷酸化的影响

J4 ›› 2010, Vol. 36 ›› Issue (6): 1021-1025.

人大肠癌lovo细胞株PARG基因沉默对蛋白激酶P38表达及其磷酸化的影响

潘娟, 王娅兰, 李巧转

（重庆医科大学分子医学与肿瘤研究中心病理学教研室|重庆400016）

收稿日期:2010-05-11 出版日期:2010-11-28 发布日期:2010-11-28
通讯作者: 王娅兰 E-mail:（Tel: 023-68485789，E-mail: wangyalan074@126.com）
作者简介:潘娟（1984-）|女| 湖北省恩施市人|在读医学硕士|主要从事大肠癌侵袭转移的研究。
基金资助:
国家自然科学基金资助课题（30870946）

Influence of silenced PARG on expression and phosphorylation of |protein kinase P38 in human colorectal carcinoma lovo cells

Pan Juan, WANG Ya-Lan, LI Qiao-Zhuai

（Department of Pathology|Molecular Medicine and Cancer Research Center|Chongqing Medical University|Chongqing 400016|China）

Received:2010-05-11 Online:2010-11-28 Published:2010-11-28

摘要/Abstract

摘要：

目的：初步探讨人大肠癌lovo细胞株聚（腺苷二磷酸核糖）水解酶［poly-（ADP-ribose）glycohydrolase，PARG］基因沉默对细胞信号通路MAPK中蛋白激酶P38表达及其磷酸化的影响，并阐明其可能机制。方法：采用PARG-shRNA慢病毒载体转染lovo细胞株并筛选出稳定沉默PARG基因的lovo细胞株，以未作处理lovo细胞为未转染组作为阳性对照，以转染未沉默PARG基因载体lovo细胞为空载体转染组作为阴性对照，以转染沉默PARG基因载体lovo细胞为目的基因转染组作为实验对照。RT-PCR检测细胞PARG mRNA表达变化，Western blotting检测PARG、聚（腺苷二磷酸核糖）聚合酶［poly-（ADP-ribose）polymerases，PARP］、P38和磷酸化P38（p-P38）表达变化。结果：RT-PCR和Western blotting结果均显示，目的基因转染组PARG表达显著降低，与未转染组比较差异有显著性（P<0.05）；Western blotting 提示，目的基因转染组PARP、P38和p-P38表达均明显低于未转染组（P<0.05）。结论：人大肠癌lovo细胞PARG基因沉默可抑制P38的表达及其磷酸化，这可能与PARG下调 PARP有关。

关键词: 大肠肿瘤；聚(腺苷二磷酸核糖)水解酶；聚(腺苷二磷酸核糖)聚合酶；P38；慢病毒载体

Abstract:

To investigate the influence of silenced poly-（ADP-ribose）glycohydrolase（PARG）on the expression and phosphorylation of protein kinase P38 in colorectal carcinoma lovo cells and clarify its mechanism. Methods Lentivirus PARG-shRNA was transfected into colorectal carcinoma lovo cells，then the lovo cells with silenced PARG was perpetually selected. Untreated lovo cells were used as positive control(untransfected group)，the lovo cells treated with empty vector were used as negative control (empty-vector-transfected group)，the lovo cells treated with PARG-shRNA were used as experimental comparison(PARG- shRNA transfected group). The expression of PARG mRNA was detected by RT-PCR. The expressions of PARG，poly-（ADP-ribose）polymerases（PARP），P38 and p-P38 protein were detected by Western blotting. Results Both RT-PCR and Western blotting results showed that the expressions of PARG in PARG-shRNA transfected group was obviously lower than that in untransfected group（P<0.05）. The expressions of PARP，P38 and p-P38 in PARG-shRNA transfected group were obviously lower than that in untransfected group（P<0.05）．
Conclusion The silenced PARG can inhibit the expression and phosphorylation of P38 in human colorectal carcinoma lovo cells. It may be related to down-regulating PARP .

Key words: colonic neoplasms；poly-（ADP-ribose）
glycohydrolase；poly-(ADP-ribose）polymerases；P38；Lentivirus-shRNA

中图分类号:

R73-3

潘娟, 王娅兰, 李巧转. 人大肠癌lovo细胞株PARG基因沉默对蛋白激酶P38表达及其磷酸化的影响[J]. J4, 2010, 36(6): 1021-1025.

Pan Juan, WANG Ya-Lan, LI Qiao-Zhuai. Influence of silenced PARG on expression and phosphorylation of |protein kinase P38 in human colorectal carcinoma lovo cells[J]. J4, 2010, 36(6): 1021-1025.