1.Role of androgen receptor in CK5 +CK8 + cells isolated from prostate cancer LNCaP cells and its regulating mechanism
Zhifang MA ; Sheng ZENG ; Haiyang KUAI ; Tiankun HANG ; Chun LIU
Cancer Research and Clinic 2021;33(1):42-47
Objective:To investigate the role of androgen receptor (AR) in CK5 +CK8 + cells isolated from prostate cancer LNCaP cells and its regulating mechanism. Methods:CK5 +CK8 + cells were isolated from LNCaP cells by using flow cytometry. Lentivirus vector carrying AR gene was transferred in CK5 +CK8 + cells. The experiments were divided into AR CK5 +CK8 + group transfering AR and V CK5 +CK8 + group transfering blank load. The expressions of AR, p-AKT and bcl-2 were tested by using Western blot assay under different concentrations of androgen (1 nmol/L and 10 nmol/L dihydrotestosterone). Methyl thiazolyl tetrazolium (MTT) assay, cell migration assay and soft agarose gel clone formation assay was used to detect the effect of AR on the biological property of CK5 +CK8 + cells. The effect of activated inhibitors such as LY 294002 (LY), γ-tocotrienol (γ-TT) and/or 5-fluorocytosine inducing AR expression (5-AZA) through AKT signal pathways on CK5 +CK8 + cells proliferation was detected by using MTT assay. Results:After AR gene was transferred into CK5 +CK8 + cells, the expression of AR was increased, while the expression of p-AKT and bcl-2 was decreased. After the treatment of 1 nmol /L dihydrotestosterone and 10 nmol/L dihydrotestosterone for 2, 4 and 6 d, the cell proliferation inhibited degree of AR CK5 +CK8 + cells was higher compared with that of V CK5 +CK8 + cells, and the difference was statistically significant (all P < 0.05). After the treatment of 1 nmol/L dihydrotestosterone and 10 nmol /L dihydrotestosterone for 3 d, the migration ability of AR CK5 +CK8 + cells was decreased compared with that of V CK5 +CK8 + cells (the number of cell migration: 54±9 vs. 113±21, 13±3 vs. 34±6), and the differences were statistically significant ( t=4.450, P<0.01; t=5.157, P<0.01).After the treatment of 1 nmol /L dihydrotestosterone and 10 nmol /L dihydrotestosterone for 3 weeks, the tumorigenic ability of AR CK5 +CK8 + cells was reduced compared with that of V CK5 +CK8 + cells (the number of clone: 39±7 vs. 105±16, 41±6 vs. 86±6), and the differences were statistically significant ( t=6.631, P<0.01; t=8.662, P<0.01). And 5 nmol /L LY + 10 nmol/L 5-AZA, 5 nmol /L LY + 5 nmol/L γ-TT, 10 nmol/L 5-AZA + 5 nmol/L γ-TT, 2.5 nmol/L LY + 5 nmol/L 5-AZA + 2.5 nmol/L γ-TT combined with 1 nmol/L dihydrotestosterone or 10 nmol/L dihydrotestosterone after the treatment of 2, 4, 6 d inhibited the proliferation of CK5 +CK8 + cells (all P < 0.05). Conclusion:AR plays an inhibitory role in CK5 +CK8 + cells isolated from prostate cancer cell line LNCaP and reduces the cell migration and tumorigenic ability through inhibiting activation of AKT-bcl-2 signal pathway.
2.Emphysematous cystitis: a case report
Hongjie LI ; Lichong YU ; Haiyang KUAI ; Tongyi LI ; Zhifang MA
Chinese Journal of Urology 2021;42(5):388-389
Emphysematous cystitis is a rare inflammation of the bladder, which can be life-threatening in severe cases. A senior gentleman with emphysematous cystitis was recently admitted to our hospital. He got bilateral renal hydronephrosis and bilateral terminal pneumatosis of the ureter. The comorbidity was prostatic hyperplasia and diabetes. After indwelling catheter , glucose levels control and anti-infection treatment for 3 days, the repeated CT showed the gas inside the bladder the end of ureter decreased significantly. The blood routine, urine routine, renal function was better than before. The patient was instructed to continue anti-infection treatment. The above laboratory examination indicators returned to normal after a week's re-examination.