| 摘要: |
| ［摘要］目的 探讨脆性基因WWOX调控胆囊癌细胞的增殖效应及其机制．方法 将前期成功构建的pcDNA3.0-WWOX重组质粒按脂质体介导转染GBC-SD细胞，同时转染空载体、脂质体为阴性对照及GBC-SD为空白对照．转染48 h后采用倒置显微镜观察各组胆囊癌细胞形态学变化，MTT、BrdU检测胆囊癌细胞增殖水平，流式细胞仪检测细胞增值周期的变化．结果 倒置显微镜观察pcDNA3.0-WWOX组GBC-SD细胞数量明显减少，悬浮细胞及细胞碎片增加，而Vector control、NC、Mock组细胞呈正常增殖状态．MTT检测显示pcDNA3.0-WWOX组GBC-SD细胞增殖于24 h、48 h、72 h、96 h、120 h低于对照组，差异有统计学意义（P＜0.05），并随时间推移，pcDNA3.0-WWOX组细胞增殖活性抑制明显．BrdU检测显示pcDNA3.0-WWOX组细胞增殖率为（0.44±0.03），对照组分别为（0.78±0.02），（0.81±0.01），（0.85±0.01），表明pcDNA3.0-WWOX组细胞增殖活性较对照组降低，差异有统计学意义（P＜0.05）．细胞周期检测发现pcDNA3.0-WWOX组G0 /G1期细胞增多，G2 /M 期和S 期细胞减少，细胞凋亡率较对照组（Vector control，NC，Mock）明显增高、增殖指数下降（P＜0.05），而对照组间差异无统计学意义（P＞0.05）．结论 过表达WWOX基因体外可有效抑制胆囊癌细胞增殖活性， WWOX可能参与胆囊癌细胞恶性生物学行为的发展过程，有望成为胆囊癌基因治疗新的潜在靶点． |
| 关键词: ［关键词］脆性基因WWOX GBC-SD 重组质粒 增殖 |
| DOI： |
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| 基金项目:［基金项目］云南省应用基础研究基金资助项目（2011FZ124） |
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| Effect of Fragile Site WWOX Gene on Regulating Proliferation of Human Gallbladder Cancer Cells in Vitro |
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WEI Dong1,2
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1.（1）Dept. of Hepatopancreatobiliary Surgery;2）Dept. of Gastrointestinal Surgery，The Second Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650101，China）
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| Abstract: |
| ［Abstract］Objective To explore the effect and mechanism of fragile site WWOX gene on regulating proliferation of gallbladder cancer cells in vitro. Methods The pcDNA3.0 - WWOX recombinant plasmid which was previous successfully built was transfected to GBC-SD cells and empty carrier by liposome medium. Liposome and GBC-SD were served as the negative control and the blank control， respectively. After 48 hours transfection， inverted microscope was used to observe the changes of gallbladder cancer cells' morphology， MTT and BrdU were used to detect the proliferation level of gallbladder cancer cells，and flow cytometry instrument was used to detect the change of the cell proliferation cycle. Results The results of inverted microscope shown: the number of GBC-SD cells in pcDNA3.0-WWOX group decreased significantly，the suspension cells and cell debris increased，while cells in the vector control，NC and Mock groups were in normal proliferation state. MTT test showed the proliferation levels of GBC-SD cells in pcDNA3.0-WWOX group was lower than those in the control group in 24 h，48 h，72 h， 96 h and 120 h， and the differences were statistically significant（P < 0.05）. The cell proliferation activity in the pcDNA3.0-WWOX group was obviously inhibited over time. BrdU detection results showed the cell proliferation rate of pcDNA3.0 - WWOX group was（0.44±0.03）， while that in the three control groups was（0.78±0.02）， （0.81±0.01）and（0.85±0.01）， respectively. It showed that cell proliferation activity in pcDNA3.0-WWOX group was lower than the control groups， and the difference was statistically significant （P < 0.05）. Cell cycle detection showed the cells increased in G0/G1 phase and decreased in G2/M and S phases of pcDNA3.0-WWOX group. The cell apoptosis rate was significantly higher and the proliferation index was significantly lower than those of the control groups （P < 0.05）. However， there were no significant differences among the three control groups（P > 0.05）. Conclusion The overexpression of WWOX gene in vitro could effectively inhibit the proliferation activity of gallbladder cancer cells. WWOX might participate in the development of the malignant biological behavior of gallbladder cancer cells. It is expected to become a new potential target for the gene therapy to gallbladder cancer. |
| Key words: ［Key words］Fragile gene WWOX GBC-SD Recombinant plasmid Proliferation |