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].Com-paring 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 Ln-cRNA TapSAKI may play a role in the deterioration of cell proliferation and apoptosis.