Effect of long non-coding RNA TapSAKI on the renal tubular epithelial cells induced by hypoxia

Abstract

Objective To study the effect of long non-coding(LncRNA) TapSAKI on HK-2 cells of renal tubular epithelial cells induced by hypoxia. Methods The cultured HK-2 cells were divided into the normal oxygen group (group A), hypoxia injury group (group B), hypoxia injury+ control siRNA group (group C), and hypoxia injury+ TapSAKI siRNA group (group D). The real-time quantitative polymerase chain reaction (qPCR) was used to detect the change in TapSAKI expression; proliferation was measured by CCK-8 assay; cell cycle and cell apoptosis were measured by flow cytometry; Western blot was used to detect cycle related CyclinD1 protein, CDK2 and apoptosis related protein Bcl-2 and Bax. Results From the result of qPCR, compared with group A, the expression of TapSAKI gene in group B increased significantly (48.92±0.31 vs.1.00±0.01, P<0.05). Compared with group C, the expression of TapSAKI gene in group D decreased significantly(4.70±0.60 vs.48.31±0.29, P<0.05). The result of CCK-8 showed that the proliferation in group B significantly decreased compared with group A (P<0.05). Compared with group C, the proliferation in group D significantly increased (P<0.05). The flow cytometer test results showed that the apoptosis rate in group B was higher than that in group A [(26.38±1.21)% vs.(6.45±0.46)%, P<0.05]. Comparing with group C, the apoptosis rate in group D decreased significantly [(10.98±0.88)% vs.(21.59±1.30)%, P<0.05]. Compared with group A, Bax expression in group B increased, and Bcl-2 expression decreased (1.304±0.082 vs.0.411±0.002, 0.390±0.007 vs.1.027±0.022, all P<0.05). In group D, the expression of Bax was lower than that in group C, and the expression of Bcl-2 increased, both of which were statistically significant(0.655±0.819 vs.1.419±0.087, 0.819±0.034 vs.0.437±0.014, all P<0.05). Compared with group A, the proportion of G0/G1 phase cells in group B significantly increased (69.82±1.14 vs.34.46±0.82, P<0.05), and the proportion of S phase cells decreased significantly (11.6±0.60 vs.42.23±1.46, P<0.05). Compared with group A, CyclinD1 and CDK2 protein expressions in group B decreased (0.659±0.062 vs.1.723±0.084, 0.414±0.015 vs.0.87±0.031, all P<0.05). Compared with group C, group D G0/G1 phase cells significantly decreased (30.77±0.33 vs.61.81±1.50, P<0.05), the proportion of S phase cells significantly increased(40.32±0.72 vs.17.92±0.71, P<0.05), CyclinD1 and CDK2 protein expressions increased (2.049±0.027 vs.0.626±0.024, 0.89±0.104 vs.0.424±0.012, all P<0.05). Conclusions Under hypoxic conditions, LncRNA TapSAKI in renal tubular epithelial cells was abundant, which may inhibit renal tubular epithelial cell proliferation and accelerate apoptosis.It is suggested that LncRNA TapSAKI may play a role in the deterioration of cell proliferation and apoptosis. Key words: Long non-coding RNA; TapSAKI; Renal tubular epithelial cell; Hypoxia