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Establishment of an Androgen-independent Prostate Cancer Cell Line Induced by the Combined of Flutamide and Androgen-free Environment
Gang Xu, Yi-Jie Mao, Bin-Hua Chen, Jing-Jing He, Zhi-Hua Tao*, Xiao-Mei Lin, Zhan-Guo Chen, Mo Shen
Center for Clinical Laboratory Diagnosis, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, China
Abstract: We established an androgen-independent prostate cancer cell line induced by the combination of flutamide and androgen-free environment after long-term culture in vivo. Cell morphological changes were traced with inverted microscope. Cell proliferation rate was measured by CCK-8 assa. Flow cytometry was used to determine the changes of cell cycle， and expression of Bcl-2 and Bax. Cell morphology of experimental group from 1 to 20 passages was leptosomic， however， it was changed into flatting and aggregation growth pattern after 30 passages， cell morphology of control group was always shuttle-like shape. Cell growth rate of experimental group LNCaP cell was shown a gradual uptrend in the whole process when cultured in androgen-free medium with the terminal concentration of flutamide of 5.0×10-6 mol/L. Flutamide and androgen-free environment was revealed to have no significant effect on the cell cycle of experimental group LNCaP cell， but the control group was found to be inhibited. Mean fluorescence intensity (MFI) of Bcl-2 in experimental group was significantly higher than control group. However， there was no significant difference in the MFI of Bax between two groups. LNCaP cell could be changed into androgen-independent prostate cancer cell after long-term induced by the combination of flutamide and androgen-free environment， and this would be a suitable cell line for the pathogenesis studies.
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