前列腺癌是一種男性常罹患的癌症，在台灣前列腺癌病人的死亡率日漸增加。越來越多研究證實前列腺癌幹細胞參與前列腺癌的形成、發展、抗藥性、轉移與復發，因此了解前列腺癌幹細胞導致前列腺癌發展及惡化的機轉，與開發針對前列腺癌幹細胞的治療方式乃成為當務之急。本實驗以人類前列腺癌PC-3細胞為對象，分選出細胞間聚集生長的holoclone細胞，與細胞分散生長的paraclone細胞。當以群落形成分析，發現holoclone細胞會形成較多聚集生長的細胞群落。在細胞球體形成實驗，也發現holoclone細胞會形成較大且數目較多的細胞球體，顯示holoclone細胞可能具有較強的自我更新能力。分析holoclone細胞的癌幹細胞標誌基因表現，發現KLF-4、Oct-4、Sox-2與FAM65B等基因的表現較高，表示holoclone細胞可能具有癌幹細胞特徵。利用分選出具有癌幹細胞特徵的holoclone細胞分析，細胞爬行與侵入能力，結果發現holoclone細胞的爬行能力較差，侵入能力無明顯變化。相較之下paraclone細胞呈現較強的細胞爬行與侵入能力。綜合以上實驗結果，本研究成功分選出細胞形態聚集生長的holoclone PC-3細胞，其具有癌幹細胞特徵並表現癌幹細胞標記基因。未來此細胞可應用於癌幹細胞特徵之研究與對抗癌幹細胞之藥物開發。 Prostate cancer is one of the most common cancer affecting males and the cancer mortality is increasing in Taiwan. Prostate cancer stem cells has been shown to be involved in the prostate cancer development, chemoresistance, and metastasis. Thus, investigating the mechanisms of prostate cancer stem cells on cancer progression and developing prostate cancer stem cells-targeted therapeutic approaches are important. In the present study, we isolated holoclone cells with close cell-cell contact morphology and paraclone cells with loose cell contact morphology from human prostate cancer PC-3 cells. PC-3 holoclone cells posses prostate cancer stem cells properties, such as self-renewal spheroid formation, colony formation, and expressing cancer stem cells marker genes KLF-4、Oct-4、Sox-2 and FAM65B. Furthermore, cell migratory and invasive activities of PC-3 holoclone cells were analyzed. Results demonstrated that PC-3 holoclone cells exhibited less migratory activity, while paraclone cells revealed higher migratory and invasive activities. Taken together, we isolated PC-3 holoclone cells in vitro with cancer stem cells charactristics and expressing cancer stem cell marker genes. The cells can be used to study the regulation of prostate cancer stem cells and develop prostate cancer stem cells-tareted therapeutic approaches.
