中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (7): 499-502.

• 论著 • 上一篇    下一篇

Rab23对鳞状细胞癌Sa3细胞增殖的抑制作用及相关机制

刘曦霖1,简强1,苗叶2,黄敏1,李承新1   

  1. 1. 第四军医大学西京皮肤医院
    2. 陕西省西安市第四军医大学西京皮肤医院
  • 收稿日期:2014-01-16 修回日期:2014-04-22 发布日期:2014-07-01
  • 通讯作者: 李承新 E-mail:chengxindrem@163.com

Inhibitory effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3 and its mechanisms

  • Received:2014-01-16 Revised:2014-04-22 Published:2014-07-01

摘要: 目的 观察Rab23分子对鳞状细胞癌(简称鳞癌)细胞Sa3增殖的影响及机制。 方法 将鳞状细胞癌Sa3细胞分为4组:正常对照组[转染绿色荧光蛋白(eGFP)]、Rab23过表达组(转染eGFP标记的Rab23过表达质粒)、Rab23沉默组(转染Rab23 shRNA质粒)、Rab23空载体组(转染空载体)。平板克隆形成实验、流式细胞仪检测Rab23过表达和沉默对Sa3细胞增殖能力的影响。Western印迹检测Rab23过表达和沉默对Sa3细胞Erk/p-Erk表达水平的变化。两组间均数比较采用t检验,多组间均数比较采用单因素方差分析,多组资料两两比较使用Bonferroni′s多重比较分析。 结果 通过构建质粒、慢病毒感染,获得Rab23过表达和Rab23沉默的稳定Sa3细胞株。平板克隆形成实验结果显示,Rab23过表达组克隆形成率为2.3% ± 0.2%,与正常对照组(3.6% ± 0.3%)相比,Sa3细胞增殖能力降低(P < 0.05),而Rab23沉默组克隆形成率(4.1% ± 0.2%)较空载体组(1.8% ± 0.0%)增殖能力明显提高(P < 0.01)。细胞周期检测显示,Rab23过表达可引起Sa3细胞G1期阻滞,Rab23过表达组增殖指数(0.581 ± 0.035)较正常对照组(0.698 ± 0.018)降低(P < 0.05),而Rab23沉默组增殖指数(0.567 ± 0.015)较空载体组(0.444 ± 0.014)明显提高(P < 0.01)。Western 印迹结果显示,与正常对照组相比,Rab23过表达组和Rab23沉默组Sa3细胞Erk表达水平无变化,但Rab23过表达组磷酸化Erk表达水平较正常对照组降低,Rab23沉默组磷酸化Erk表达水平较空载体组升高。 结论 Rab23对鳞癌Sa3细胞增殖起抑制作用,这种抑制作用可能与Erk通路有关。

关键词: Rab23, 慢病毒感染, 癌,鳞状细胞, 细胞系,肿瘤, 细胞增殖

Abstract: Liu Xilin, Jian Qiang, Miao Ye, Huang Min, Li Chengxin. Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi′an 710032, China Corresponding author: Li Chengxin, Email: chengxinderm@163.com 【Abstract】 Objective To evaluate the effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3, and to investigate its mechanisms. Methods Cultured Sa3 cells were classified into four groups: normal control group transfected with green fluorescent protein (GFP), Rab23-overexpressing group transfected with a GFP-labelled Rab23-overexpressing plasmid, Rab23-silencing group transfected with a plasmid carrying a Rab23-targeting shRNA, empty vector group transfected with an empty vector. After additional culture for different durations, plate colony formation assay and flow cytometry were performed to evaluate the proliferative activity of Sa3 cells, and Western blot was conducted to detect the expression of Erk/phosphorylated-Erk in Sa3 cells. Statistical analysis was carried out by t test, one-way analysis of variance and Bonferroni′s multiple comparison test. Results Stable Sa3 cell lines with overexpression or silencing of Rab23 were established by plasmid construction and lentivirus-mediated transfection. The plate colony formation assay showed that the colony formation rate was significantly lower in the Rab23-overexpressing group than in the normal control group (2.3% ± 0.2% vs. 3.6% ± 0.3%, P < 0.05), but higher in the Rab23-silencing group than in the empty vector group (4.1% ± 0.2% vs. 1.8% ± 0.03%, P < 0.01). Rab23 overexpression induced G1 phase arrest in Sa3 cells. The proliferation index was significantly decreased in the Rab23-overexpressing group compared with the normal control group(0.581 ± 0.035 vs. 0.698 ± 0.018, P < 0.05), but increased in the Rab23-silencing group compared with the empty vector group (0.567 ± 0.015 vs. 0.444 ± 0.014, P < 0.01). As Western blot showed, there were no significant changes in the expression of Erk in the Rab23-silencing or -overexpressing group compared with the normal control group, whereas the expression of p-Erk was attenuated in the Rab23-overexpressing group compared with the normal control group, but enhanced in the Rab23-silencing group compared with the empty vector group. Conclusions Rab23 could inhibit the proliferation of Sa3 cells, which may be associated with the Erk pathway.

Key words: Rab23, Lentivirus infections, Carcinoma, squamous cell, Cell line, tumor, Cell proliferation

中图分类号: 

  • R75

引用本文

刘曦霖 简强 苗叶 黄敏 李承新. Rab23对鳞状细胞癌Sa3细胞增殖的抑制作用及相关机制[J]. 中华皮肤科杂志, 2014,47(7):499-502. doi: