Gossypol inhibits invasion potential of human cervical carcinoma cells in vitro and in vivo

• Main Authors: MENG-HSUAN LIN, 林孟璇
• Other Authors: 謝易修
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/50338325352199908120

碩士 === 中山醫學大學 === 生化暨生物科技研究所 === 102 === The metastasis of cervical cancer is the most prevalent cause of patient death, and various treatment strategies have targeted the prevention of the occurrence of metastasis. Gossypol is a natural phenol derived from the cotton plant and is a phenolic aldehyde that permeates cells and acts as an inhibitor for several dehydrogenase enzymes. Gossypol is reported to exhibit antifertility, antioxidant, anticancer, antivirus, and antimicrobial properties. However, the effects of gossypol on cancer invasion and tumor growth of the human cervical carcinoma remain unclear. In this study, anti-metastasis, anti-angiogenesis, anti-tumor effects and reversion of EMT were investigated using gossypol on two cervical cancer call lines (SiHa and HeLa). SiHa and HeLa cells were treated with gossypol to determine the effect on cell proliferation by MTT assay. To examine the inhibitory effect on the cell invasion and migration, Boyden chamber invasion assay and wound healing assay were performed. We examined the effect of gossypol on factors of cancer metastasis and epithelial to mesenchymal transition (EMT) by Western Blot. Condition media of cells were subjected to gelatin zymography and casein zymography to investigate the expression of matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (u-PA). The molecular mechanisms of gossypol mediated cancer metastasis and EMT were further investigated by Western blotting analysis including MAPK, FAK/Src and PI3K/Akt signaling pathways. Gossypol inhibits the viability of human cervical carcinoma cells in a dose- and time-dependent manner. By cell invasion assay, gossypol reduces the invasion of SiHa and HeLa cells in a concentration-dependent manner. Zymography assay has shown gossypol reduces the activities of u-PA and MMP-2. Gossypol inhibits cell migration of SiHa and HeLa cells. Gossypol was sufficient to inhibit TGF-β1-induced p-ERK1/2 and p-Smad3 expression and increase TGF-β1-reduced E-cadherin (MET marker) expression in SiHa cells. Gossypol reverses the TGF-β1 induced conformation change (mesenchymal type) using immunofluorescence assay. Finally, gossypol was evidenced by its inhibition on the tumor growth and metastasis of SiHa cells via cancer cell xenografted nude mice and tail vein injection scid mice mode, respectively. Taken together, these results suggested that gossypol could reduce the invasion and reverse EMT in human cervical cancer cells.