[靶向polo样激酶1的小RNA体外及体内对胃癌细胞生长抑制作用的研究]

[Study of growth inhibition of gastric cancer cells by sRNA targeting polo like kinase 1 in vitro and vivo].

作者信息

Lan Bin, Liu Bing-ya, Cheng Xue-hua, Qu Ying, Zhang Xiao-qing, Cai Qu, Dai Qi-bao, Zhu Zheng-Gang

机构信息

Department of Surgery, Shanghai Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai 200025, China.

出版信息

Zhonghua Wai Ke Za Zhi. 2006 Jan 1;44(1):40-4.

PMID:16620646

Abstract

OBJECTIVE

To observe the effect of polo like kinase 1 (plk1) gene depletion on the growth of gastric cancer cell line-MKN45 cells in vitro and vivo and discuss the feasibility and effectiveness of arranging plk1 as gene therapeutic target for gastric cancer.

METHODS

The plk1 expression of MKN45 cells was inhibited by RNA interference (RNAi). The plk1 mRNA and protein level were measured by real-time quantitative PCR and western blotting, and the change of cell cycle distribution and apoptosis rate were detected by flow-cytometry, and the MKN45 cells proliferation was measured by MTT method. MKN45 cells treated with plk1 siRNA were transplanted subcutaneously in nude mice and their tumorgenesis ability were observed, the plk1 protein levels of the samples from nude mice in different groups were compared.

RESULTS

After treatment with plk1 siRNA, plk1 mRNA and protein level decreased obviously in certain time, more MKN45 cells accumulated at G(2)/M (P < 0.05). Apoptosis rate of MKN45 cells treated with plk1 siRNA was higher than that of control cells at 48 h and 72 h (P < 0.05), and MKN45 cells proliferated slowly than control groups (P < 0.05), while the tumorgenesis ability obviously decreased, but the plk1 protein levels of the samples from nude mice in different groups were not different.

CONCLUSIONS

siRNA targeting plk1 can inhibit the proliferation of MKN45 cells in vitro and vivo. Plk1 may be a novel therapeutic target for gastric cancer.

摘要

目的

观察 polo 样激酶 1（plk1）基因缺失对胃癌细胞系-MKN45 细胞体外及体内生长的影响，并探讨将 plk1 作为胃癌基因治疗靶点的可行性和有效性。

方法

采用 RNA 干扰（RNAi）抑制 MKN45 细胞中 plk1 的表达。通过实时定量 PCR 和蛋白质免疫印迹法检测 plk1 mRNA 和蛋白水平，采用流式细胞术检测细胞周期分布及凋亡率的变化，用 MTT 法检测 MKN45 细胞的增殖情况。将经 plk1 siRNA 处理的 MKN45 细胞皮下接种于裸鼠，观察其成瘤能力，比较不同组裸鼠样本中 plk1 蛋白水平。

结果

用 plk1 siRNA 处理后，在一定时间内 plk1 mRNA 和蛋白水平明显下降，更多 MKN45 细胞积聚于 G(2)/M 期（P < ０.０５）。在 48 h 和 72 h 时，经 plk1 siRNA 处理的 MKN45 细胞凋亡率高于对照细胞（P < ０.０５），且 MKN45 细胞增殖比对照组缓慢（P < ０.０５），而成瘤能力明显降低，但不同组裸鼠样本中 plk1 蛋白水平无差异。