Xiong Zhoufang, Hu Sha, Wang Zehua
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Jun;30(3):365-9. doi: 10.1007/s11596-010-0358-z. Epub 2010 Jun 17.
The growth-inhibiting and apoptosis-inducing effects of WW domain-containing oxidoreductase (WWOX) gene on ovarian cancer cell line A2780 were investigated. The full length cDNA of human WWOX gene was amplified from normal human ovary tissues. The correct cDNA of full length WWOX was subcloned into eukaryocytic expression vector pCMV. After introduction of WWOX gene into cancer cells with liposome, the WWOX mRNA and protein level in the cancer cells were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunoblotting. The growth activities of cancer cells were detected by Trypan blue staining. The clone formation assay in soft agar was employed to observe the proliferation of the cancer cells. Apoptosis was examined by DNA ladder and acridine orange-ethidium bromide fluorescent staining. The results showed that 72 h after WWOX gene transfection, the WWOX expression was increased significantly (P<0.01). The growth of ovarian cancer cells was decreased by 16.41% to 38.49% (P<0.01). The clone formation abilities were reduced (P<0.01). Some cancer cells presented the characteristic morphological changes of apoptosis with obvious ladder bands on electrophoresis. The apoptosis rate was (20.7+/-6.0)% (P<0.01). It was concluded that over-expression of WWOX gene could induce apoptosis and inhibit the growth of ovarian cancer cells, which might be potentially useful in the gene therapy of ovarian cancers.
研究了含WW结构域氧化还原酶（WWOX）基因对卵巢癌细胞系A2780的生长抑制和凋亡诱导作用。从正常人卵巢组织中扩增出人WWOX基因的全长cDNA。将正确的全长WWOX cDNA亚克隆到真核表达载体pCMV中。用脂质体将WWOX基因导入癌细胞后，通过逆转录聚合酶链反应（RT-PCR）和免疫印迹检测癌细胞中WWOX mRNA和蛋白水平。通过台盼蓝染色检测癌细胞的生长活性。采用软琼脂克隆形成试验观察癌细胞的增殖情况。通过DNA梯带和吖啶橙-溴化乙锭荧光染色检测凋亡情况。结果显示，WWOX基因转染72小时后，WWOX表达显著增加（P<0.01）。卵巢癌细胞的生长下降了16.41%至38.49%（P<0.01）。克隆形成能力降低（P<0.01）。一些癌细胞呈现出凋亡的特征性形态变化，电泳上有明显的梯带。凋亡率为（20.7±6.0）%（P<0.01）。结论是，WWOX基因的过表达可诱导卵巢癌细胞凋亡并抑制其生长，这可能在卵巢癌的基因治疗中具有潜在应用价值。
