[WWOX基因对卵巢癌细胞黏附与附着的影响]

[Effect of WWOX gene on the attachment and adhesion of ovarian cancer cells].

作者信息

Zhang Jie-qing, Li Li, Song Hong-lin, Paige Adam, Gabra Hani

机构信息

Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, China.

出版信息

Zhonghua Fu Chan Ke Za Zhi. 2009 Jul;44(7):529-32.

PMID:19957554

Abstract

OBJECTIVE

To explore the relationship between WWOX gene and attachment and adhesion of ovarian cancer.

METHODS

The expression of WWOX mRNA was detected by RT-PCR, the expression of the WWOX protein was evaluated by western blot in WWOX-transfected PEO1 cells (H6, H7, H8 cell) and vector-transfected control cells (vec-1, vec-2 cell). Attachment assay was used to assess the adhesion of the transfection in PEO1 cells via culturing the cells on the pre-coated fibronectin wells. RNA interference (RNAi) was used to knockdown the endogenous expression of WWOX in the A2780 ovarian cancer cell line by liposome. Attachment assay was detected the adhesion to fibronectin after gene silencing.

RESULTS

RT-PCR showed that expression of mRNA WWOX in exon9 was in all transfection cells (H6, H7, H8, vec-1, vec-2 cell). Western blot showed that expression of WWOX protein was in the WWOX-transfected cells (H6, H7, H8 cell), but not in the vector-transfected cells (vec-1, vec-2 cell). Attachment assay showed that H6, H7, H8 cell (0.098 +/-0. 003, 0.091 +/- 0.004, 0.099 +/- 0.003) adhered more slowly to fibronectin than vec-1, vec-2 cell (0.185 +/- 0.003, 0.175 +/- 0.006) and non-transfected PEO1 cell (0.211 +/- 0.007), and demonstrated significantly reduced adhesion after 2 hours (P < 0.01). A2780 adhesive cells that WWOX gene be knockdown was 0.059 +/- 0.005, adhered more significantly rapid than those untreated cells that was 0.029 +/- 0.003 after treated 30 minutes (P < 0.05).

CONCLUSIONS

WWOX gene can suppress adhesion to fibronectin in ovarian cancer cells. This suggests an important role for loss of WWOX gene in promoting attachment and adhesion of ovarian cancer cells on loco-regional peritoneum, and further resulting in enhancing loco-regional peritoneal tumor invasiveness and spread.

摘要

目的

探讨WWOX基因与卵巢癌黏附及附着的关系。

方法

采用逆转录聚合酶链反应（RT-PCR）检测WWOX转染的PEO1细胞（H6、H7、H8细胞）及载体转染的对照细胞（vec-1、vec-2细胞）中WWOX mRNA的表达，采用蛋白质免疫印迹法（western blot）评估WWOX蛋白的表达。通过将细胞接种于预包被纤连蛋白的孔板中，采用黏附实验评估PEO1细胞转染后的黏附情况。利用脂质体介导的RNA干扰（RNAi）技术敲低A2780卵巢癌细胞系中WWOX的内源性表达。基因沉默后，采用黏附实验检测细胞对纤连蛋白的黏附情况。

结果

RT-PCR结果显示，所有转染细胞（H6、H7、H8、vec-1、vec-2细胞）中外显子9的WWOX mRNA均有表达。western blot结果显示，WWOX转染细胞（H6、H7、H8细胞）中有WWOX蛋白表达，而载体转染细胞（vec-1、vec-2细胞）中无表达。黏附实验显示，H6、H7、H8细胞（0.098±0.003、0.091±0.004、0.099±0.003）对纤连蛋白的黏附速度慢于vec-1、vec-2细胞（0.185±0.003、0.175±0.006）及未转染的PEO1细胞（0.211±0.007），且在2小时后黏附明显降低（P<0.01）。WWOX基因敲低的A2780黏附细胞为0.059±0.005，在处理30分钟后，其黏附速度明显快于未处理细胞（0.029±0.003）（P<0.05）。