The Effect Of Urothelial Carcinoma Associated 1 On The Akt Signal Pathway Of 3T3-L1 Adipocyte

Objective To observe the effect of Long noncoding RNA(lncRNA) urothelial carcinoma associated 1 (UCAl) on the Akt signal pathway of 3T3-L1 adipocyte and glucose uptake activity.Methods UCAl overexpression plasmid was constructed, while the specific siRNAs of UCAl were selected, then UCAl was overexpressed or silenced by plasmids or specific siRNAs transfection in 3T3-L10.Then the effect of UCAl on the Akt signal pathway of 3T3-L1 adipocyte was measured by Western blot. The transcription activity and mRNA level of peroxisome proliferator activated receptor y was detected by dual luciferase reporter assay and real-time quantity PCR. And 2-deoxy-3H-D-glucose incorporation assay was performed to measure the glucose uptake activity of 3T3-L1 adipocyte.SPSS12.0 statistical software was used for statistical analysis. Experimental data were showed in average ± standard deviation, and t test was performed in comparison between the two groups, while single factor variance analysis for multiple groups comparison.Results The mRNA level of UCAl was gradually elevated in the differentiation process of 3T3-L1 from pre-adipocyte to mature adipocyte, up to 3.48 ± 0.09 times, t=12.093, P<0.01.Overexpression of UCAl in 3T3-L1 adipocyte could enhance the phosphorylation of Akt and FOXO1:p-AKT(S473):Control vs UCAl,1103±42 vs 233859, t=16.390, P<0.01; p-AKT(T308):Control vs UCA1,1524±34 vs 3565±45, t=13.025, P<0.01; p-FOXO1:Control vs UCA1,912±21 vs 2190±31, t=10.544, P<0.01;and down-regulation of UCA1 could reverse the phosphorylation of Akt and FOXO1:p-AKT(S473):si-scramble vs si-UCA1, 1090±22 vs 540±34, t=11.045, P<0.01; p-AKT(T308):si-scramble vs si-UCA1,1409±26 vs 805±18, t=9.527, P<0.01; p-FOXO1:si-scramble vs si-UCA1,2444±29 vs 459±31, t=16.899, P<0.01. Overexpression of UCA1 could also enhance the glucose uptake activity, up to 2.21±0.09 times, t=5.922, P<0.05, and down-regulation of UCA1 could reduce the glucose uptake activity by 66%,t=5.557, P<0.05. Overexpression of UCA1 could enhance the glucose uptake activity corporately with the stimulation of insulin, up to 3.25±0.12 times, t=5.709, P<0.05, and down-regulation of UCAl could reduce the glucose uptake activity by77%, t=6.567, P<0.05.Conclusion UCA1 could activate the Akt signal pathway of 3T3-L1 adipocyte, improve the transcription activity and protein expression of PPARy, and enhance the glucose uptake activity.