Cloning of WWOX gene and its growth-inhibiting effects on ovarian cancer cells.
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.
Xiong
Zhoufang
Z
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. xiaoxiongmaomi@yahoo.com.cn
Hu
Sha
S
Wang
Zehua
Z
D017209:Apoptosis / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D000074584:WW Domain-Containing Oxidoreductase
D012333:RNA, Messenger; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human