Liu Ran-lu, Zhang Zhi-hong, Zhao Wei-ming, Wang Meng, Qi Shi-yong, Li Jing, Zhang Yue, Li Sheng-zhi, Xu Yong
Department of Urology, Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin 300211, China.
Chin Med J (Engl). 2008 Feb 20;121(4):299-304.
Nucleostemin is essential for the proliferation and survival of stem and cancer cells, but it is unknown whether this newly identified molecule is involved in prostate cancer pathogenesis.
Total RNA and protein were extracted from prostate cancer tissues and PC-3, LNCap and DU145 cell lines. The nucleostemin mRNA and protein expression were measured by RT-PCR and Western blot. Immunohistochemistry was also used to detect the nucleostemin protein expression in prostate cancer tissues and PC-3 cells. A nucleostemin specific, short hairpin RNA, expression plasmid was used to transfect PC-3 cells. The changes of nucleostemin gene were detected and the proliferative capacity of the cells was determined.
Nucleostemin was highly expressed in prostate cancer tissues and cell lines. Nucleostemin expression level in the silencer group PC-3 cells remarkably reduced. The proliferation rate of silencer group PC-3 cells decreased and the percentage of G1 stage cells increased. The neoplasm forming capacity in nude mice of the silencer group PC-3 cells decreased significantly.
Nucleostemin is highly expressed in prostate cancer tissues and cell lines. The proliferative capacity of PC-3 cells is remarkably reduced after silencing nucleostemin gene expression.
核仁素对干细胞和癌细胞的增殖及存活至关重要,但这种新发现的分子是否参与前列腺癌的发病机制尚不清楚。
从前列腺癌组织以及PC-3、LNCap和DU145细胞系中提取总RNA和蛋白质。通过逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测核仁素mRNA和蛋白质表达。免疫组织化学也用于检测前列腺癌组织和PC-3细胞中的核仁素蛋白表达。使用核仁素特异性短发夹RNA表达质粒转染PC-3细胞。检测核仁素基因的变化并测定细胞的增殖能力。
核仁素在前列腺癌组织和细胞系中高表达。沉默组PC-3细胞中的核仁素表达水平显著降低。沉默组PC-3细胞的增殖率下降,G1期细胞百分比增加。沉默组PC-3细胞在裸鼠中的成瘤能力显著降低。
核仁素在前列腺癌组织和细胞系中高表达。沉默核仁素基因表达后,PC-3细胞的增殖能力显著降低。