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Establishment Of Androgen-independent Human Prostate Cancer LNCaP Cell Model And Study On Molecular Alterations

Posted on:2006-10-16Degree:DoctorType:Dissertation
Country:ChinaCandidate:H M YeFull Text:PDF
GTID:1104360155450758Subject:Surgery
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Objective To establish and appraise androgen independent human prostate cancer LNCaPcell line model. Methods Culture LNCaP cell in the serous circumstance of long-term absence of hormone and gain LNCaP cell line which can accommodate the circumstance without hormone. Verify the reproductive activity, PSA expression and secreted PSA level of LNCaP-AI cell line in the no hormone circumstance using MTT, immun fluorescence and RT-PCR methods . Detect AR, NKX3.1 gene and Bcl-2, Bax, Akt, p-Akt protein expression in LNCaP-AI cell line using RT-PCR and western blot methods. Study the affect of hormonewithdrawal to LNCaP cell cycle by Flow Cytometry. Results LNCaP cells gradually suited for the no hormone circumstance and began to reproduce after three months .At this time the cells become androgen independent LNCaP-AI cell line.LNCaP-AI cells reproduced rapidly in the no hormone circumstance and could secreted PSA .However, PSA expression of LNCaP-AI cell was suppressed according to LNCaP cell. AR,NKX3.1 gene and Bax, Akt protein express level between LNCaP cell and LNCaP-AI cell was no significant. Bcl-2 and p-Akt increased inLNCaP-AI cell .Androgen withdrawal might drive LNCaP cell into G1 stage. Conclusions Androgen independent LNCaP-AI cell line was an ideal model to study pathogenesis of AIPC and could imitate the process from ADPC to AIPC. Androgen withdrawal may drive LNCaP cells into Gl stage. The increase in the expression of Bcl-2 and p-Akt and the increase in the ratio of Bcl-2/Bax may be the important reasons that androgen-stimulated PCa gained the ability of resistance to androgen ablation.
Keywords/Search Tags:prostate, carcinoma, androgen-independent, LNCaP model, mechanism, Bcl-2, Bax, p-Akt, Akt
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