CAJ | 학술논문

目的：观察长链非编码RNA（lncRNA）尿路上皮癌相关基因1（UCA?1）对3T3?L1脂肪细胞丝氨酸/苏氨酸蛋白激酶（Akt）信号通路及糖代谢的影响。方法通过转染UCA?1过表达质粒或UCA?1特异性siRNA干预3T3?L1脂肪细胞中UCA?1的水平，western blotting检测Akt信号通路相关蛋白磷酸化水平的变化，通过2?脱氧?3H?D?葡萄糖掺入法检测细胞的葡萄糖摄取能力。两组之间比较用t检验，多组之间比较采用单因素方差分析。结果随着3T3?L1前脂肪细胞诱导分化为成熟的脂肪细胞，UCA?1的mRNA水平逐渐升高，第6天时为诱导前的（3.48±0.09）倍（t=12.093，P<0.01）；过表达UCA?1可增加3T3?L1脂肪细胞中Akt及FOXO1的磷酸化水平[分别为p?AKT（S473）：2338±59比1103±42；p?AKT（T308）：3565±45比1524±34，p?FOXO1：2190±31比912±21，t=16.390、13.025、10.544，均P<0.01]；而利用特异性siRNA沉默UCA?1，可降低细胞中Akt及FOXO1的磷酸化水平[分别为p?AKT（S473）：540±34比1090±22；p?AKT（T308）：805±18比1409±26；p?FOXO1：459±31比2444±29，t=11.045、9.527、16.899，均P<0.01]；未给予胰岛素刺激时，过表达UCA?1可增加细胞的葡萄糖摄取能力（2.21±0.09）倍（t=5.922，P<0.05），沉默UCA?1可使细胞的葡萄糖摄取能力降低66%以上（t=5.557，P<0.05）；而给予胰岛素刺激时，过表达UCA?1可增加细胞的葡萄糖摄取能力（3.25±0.12）倍（t=5.709，P<0.05），沉默UCA?1可使细胞的葡萄糖摄取能力降低77%以上（t=6.567，P<0.05）。结论 UCA?1可激活3T3?L1脂肪细胞Akt信号通路，并促进其葡萄糖摄取能力。
목적：관찰장련비편마RNA（lncRNA）뇨로상피암상관기인1（UCA?1）대3T3?L1지방세포사안산/소안산단백격매（Akt）신호통로급당대사적영향。방법통과전염UCA?1과표체질립혹UCA?1특이성siRNA간예3T3?L1지방세포중UCA?1적수평，western blotting검측Akt신호통로상관단백린산화수평적변화，통과2?탈양?3H?D?포도당참입법검측세포적포도당섭취능력。량조지간비교용t검험，다조지간비교채용단인소방차분석。결과수착3T3?L1전지방세포유도분화위성숙적지방세포，UCA?1적mRNA수평축점승고，제6천시위유도전적（3.48±0.09）배（t=12.093，P<0.01）；과표체UCA?1가증가3T3?L1지방세포중Akt급FOXO1적린산화수평[분별위p?AKT（S473）：2338±59비1103±42；p?AKT（T308）：3565±45비1524±34，p?FOXO1：2190±31비912±21，t=16.390、13.025、10.544，균P<0.01]；이이용특이성siRNA침묵UCA?1，가강저세포중Akt급FOXO1적린산화수평[분별위p?AKT（S473）：540±34비1090±22；p?AKT（T308）：805±18비1409±26；p?FOXO1：459±31비2444±29，t=11.045、9.527、16.899，균P<0.01]；미급여이도소자격시，과표체UCA?1가증가세포적포도당섭취능력（2.21±0.09）배（t=5.922，P<0.05），침묵UCA?1가사세포적포도당섭취능력강저66%이상（t=5.557，P<0.05）；이급여이도소자격시，과표체UCA?1가증가세포적포도당섭취능력（3.25±0.12）배（t=5.709，P<0.05），침묵UCA?1가사세포적포도당섭취능력강저77%이상（t=6.567，P<0.05）。결론 UCA?1가격활3T3?L1지방세포Akt신호통로，병촉진기포도당섭취능력。
Objective To observe the effect of long noncoding RNA(lncRNA)-urothelial carcinoma associated 1 (UCA-1) on the Ser/Thr protein kinase (Akt) signal pathway in 3T3?L1 adipocyte and glucose uptake activity. Methods UCA-1 was overexpressed or silenced by plasmids or specific siRNAs transfection into 3T3?L1.Then the effect of UCA-1 on the Akt signal pathway in 3T3?L1 adipocyte was measured by western blotting. And 2?deoxy?3H?D?glucose incorporation assay was performed to measure the glucose uptake activity in 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 UCA-1 was gradually elevated in the differentiation process of 3T3?L1 from pre?adipocyte to mature adipocyte, up to (3.48 ± 0.09) folds, t=12.093, P<0.01 on Day 6, compared to pre-adipocyte. Overexpression of UCA-1 in 3T3?L1 adipocyte could enhance the phosphorylation of Akt and FOXO 1(p?AKT(S473):1 103±42 vs 2 338±59, t=16.390, P<0.01;p?AKT(T308):1 524±34 vs 3 565±45, t=13.025, P<0.01; p?FOXO1: 912 ± 21 vs 2 190 ± 31, t=10.544, P<0.01). Specific siRNA can down?regulate UCA-1, reverse the phosphorylation of Akt and FOXO1 (p?AKT(S473):1 090±22 vs 540±34, t=11.045, P<0.01;p?AKT(T308):1 409 ± 26 vs 805 ± 18, t=9.527, P<0.01;p?FOXO1:2 444 ± 29 vs 459 ± 31, t=16.899, P<0.01).Overexpression of UCA-1 could also enhance the glucose uptake activity, up to (2.21±0.09) folds, t=5.922, P<0.05, and down?regulation of UCA-1 could reduce the glucose uptake activity by 66%,t=5.557, P<0.05. Overexpression of UCA-1 could enhance the glucose uptake activity corporately with the stimulation of insulin, up to (3.25 ± 0.12) folds, t=5.709, P<0.05, and down?regulation of UCA-1 could reduce the glucose uptake activity by 77%, t=6.567, P<0.05. Conclusion UCA-1 could activate the Akt signal pathway in 3T3?L1 adipocyte and enhance the glucose uptake activity.