Palbociclib induces cell cycle arrest and senescence of human renal tubular epithelial cells / 南方医科大学学报

OBJECTIVE@#To investigate the effect of palbociclib on cell cycle progression and proliferation of human renal tubular epithelial cells.@*METHODS@#Human renal tubular epithelial cell line HK-2 was treated with 1, 5, 10, and 20 μmol/L of palbociclib, and the changes in cell proliferation and viability were examined by cell counting and CCK8 assay. EDU staining was used to assess the proliferation of HK-2 cells following palbiciclib treatment at different concentrations for 5 days. The effect of palbociclib on cell cycle distribution of HK-2 cells was evaluated using flow cytometry. SA-β-Gal staining and C12FDG senescence staining were used to detect senescence phenotypes of HK-2 cells after palbociclib treatment at different concentrations for 5 days. The relative mRNA expression levels of P16, P21, and P53 and the genes associated with senescence-related secretion phenotypes were detected by RT-PCR, and the protein expressions of P16, P21 and P53 were detected by Western blotting.@*RESULTS@#Palbociclib inhibited HK-2 cell proliferation and induced cell cycle arrest in G1 phase. Compared with the control cells, HK-2 cells treated with high-dose (10 μmol/L) palbociclib exhibited significantly suppressed cell proliferation activity, and the inhibitory effect was the most obvious on day 5 (@*CONCLUSIONS@#Palbociclib induces HK-2 cell senescence by causing cell growth arrest and delaying cell cycle progression.

Role of TLR2 and TLR4 in Mycobacterium bovis Bacillus Calmette-Guérin-induced injury in renal tubule epithelial cells / 中国病理生理杂志

AIM:To explore the effect of Toll-like receptor ( TLR) 2 and TLR4 in Mycobacterium bovis Bacil-lus Calmette-Guérin (BCG)-induced human proximal renal tubule epithelial cell (HK-2) injury.METHODS:HK-2 cells were stimulated by BCG, and the expression of TLR2, TLR4, chemokine (C-X3-C motif) ligand 1 (CX3CL1) and trans-forming growth factor beta 1 ( TGF-β1 ) was detected by quantitative real-time PCR and Western blotting .TLR2 monoclonal antibody and TLR4 inhibitor were used to treat the HK-2 cells 1 h before BCG stimulation.The expression of CX3CL1 and TGF-β1 was evaluated by quantitative real-time PCR and Western blotting .RESULTS: BCG increased the expression of TLR2, TLR4, CX3CL1 and TGF-β1 in the HK-2 cells.Additionally, the expression of CX3CL1 and TGF-β1 was inhibited partly by TLR2 monoclonal antibody or TLR4 inhibitor.CONCLUSION:BCG is able to increase the production of TLR 2, TLR4, CX3CL1 and TGF-β1 in the HK-2 cells.TLR2 and TLR4 signaling pathways play important roles in tubule epitheli-al cell injury induced by BCG .