CAJ | 학술논문

目的：探讨醉茄素A（ WFA）对非小细胞肺癌（ NSCLC） A549细胞增殖、凋亡及PI3K/Akt信号通路的影响。方法采用、2.5、5.0、10.0、20.0μmol/L WFA处理A549细胞，采用四甲基偶氮唑盐（ MTT）比色法检测上述浓度处理、48、72和h的细胞增殖抑制率，Hoechst染色和磷酯酰丝氨酸结合蛋白-异硫氢酸荧光素/碘化丙啶双染法（ Annexin V-FITC/PI）检测各浓度组h的细胞凋亡情况，流式细胞仪检测各浓度组h的细胞周期分布情况，免疫印迹检测各浓度组h凋亡相关基因（ Bcl-2、Bax和Cleaved caspase-3）和PI3K/Akt信号通路重要蛋白Akt及其磷酸化形式p-Akt的蛋白水平。结果 WFA可抑制细胞增殖，并呈剂量和时间依赖性（ P＜0.05）；0、2.5、5.0、10.0、20.0μmol/L WFA作用h后A549细胞的凋亡指数分别为.75±0.64、4.61±1.36、9.75±2.78、12.92±3.42和.68±4.31，组间差异有统计学意义（ P＜0.05）。除.5μmol/L外，其余浓度组的早、晚期凋亡率、凋亡促进基因（ Bax和Cleaved caspase-3）水平及G0/G1期细胞比例均高于μmol/L，凋亡抑制基因Bcl-2水平及S期和G2/M期细胞比例均低于μmol/L（ P＜0.05）；2.5、5.0、10.0、20.0μmol/L的组间差异有统计学意义（ P＜0.05）。随着浓度升高，p-Akt/Akt值呈降低趋势，差异有统计学意义（ P＜0.05）。结论 WFA能够抑制A549细胞的增殖及凋亡，可能通过抑制PI3K/Akt通路激活实现。
목적：탐토취가소A（ WFA）대비소세포폐암（ NSCLC） A549세포증식、조망급PI3K/Akt신호통로적영향。방법채용、2.5、5.0、10.0、20.0μmol/L WFA처리A549세포，채용사갑기우담서염（ MTT）비색법검측상술농도처리、48、72화h적세포증식억제솔，Hoechst염색화린지선사안산결합단백-이류경산형광소/전화병정쌍염법（ Annexin V-FITC/PI）검측각농도조h적세포조망정황，류식세포의검측각농도조h적세포주기분포정황，면역인적검측각농도조h조망상관기인（ Bcl-2、Bax화Cleaved caspase-3）화PI3K/Akt신호통로중요단백Akt급기린산화형식p-Akt적단백수평。결과 WFA가억제세포증식，병정제량화시간의뢰성（ P＜0.05）；0、2.5、5.0、10.0、20.0μmol/L WFA작용h후A549세포적조망지수분별위.75±0.64、4.61±1.36、9.75±2.78、12.92±3.42화.68±4.31，조간차이유통계학의의（ P＜0.05）。제.5μmol/L외，기여농도조적조、만기조망솔、조망촉진기인（ Bax화Cleaved caspase-3）수평급G0/G1기세포비례균고우μmol/L，조망억제기인Bcl-2수평급S기화G2/M기세포비례균저우μmol/L（ P＜0.05）；2.5、5.0、10.0、20.0μmol/L적조간차이유통계학의의（ P＜0.05）。수착농도승고，p-Akt/Akt치정강저추세，차이유통계학의의（ P＜0.05）。결론 WFA능구억제A549세포적증식급조망，가능통과억제PI3K/Akt통로격활실현。
Objective To explore the influences of withaferin A ( WFA) on proliferation, apoptosis and PI3K/Akt signaling pathway of non-small cell lung cancer (NSCLC) A549 cell. Methods The A549 cells were treated with different concentrations of WFA ( 0, 2.5, 5.0, 10.0, 20.0 μmol/L) . The MTT assay was used to measure the proliferation inhibition rates at 24, 48, 72 and 96h treated with different concentrations of WFA. The Hoechst staining and Annexin V-FITC/PI double staining were employed to de-tect cell apoptosis after treatment with different concentrations of WFA. The cycle distribution at 48h after treatment with WFA was de-tected by flow cytometry. The Western blotting was used to measure the protein levels of apoptosis-related genes ( Bcl-2, Bax and Cleaved caspase-3) , Akt and its phosphorylated form ( P-Akt) . Results WFA could increase the proliferation inhibition rates in a dose-and time-dependent manner( P<0.05) . The apoptotic indices of A549 cells after treatment with different concentrations of WFA (0, 2.5, 5.0, 10.0, 20.0 μmol/L) were 2.75±0.64, 4.6±1.36, 9.75±2.78, 12.92±3.42 and 18.68±4.31 with significant differences ( P<0.05) . In addition to 2.5μmol/L group, the early and late apoptosis rates, protein levels of apoptosis-promoting genes ( Bax and Cleaved caspase-3) and the cell proportion in G0/G1 phase of the remaining concentration groups were higher than those of 0μmo/L group(P<0.05).The p-Akt/Akt value decreased with increasing concentration and the differences were statistically significant among concentrations ( P<0.05) . Conclusion WFA can inhibit the proliferation and apoptosis of A549 cell possibly by inhibiting the activation of PI3K/Akt pathway activation.