聂佳丽,殷海棠,文志鹏,郑志昌,李明,李琴,樊兴华,赵青青,张彦燕,杨继红
NIE Jiali,YIN Haitang,WEN Zhipeng,ZHENG Zhichang,LI Ming,LI Qin,FAN Xinghua,ZHAO Qingqing,ZHANG Yanyan,YANG Jihong

• 1:贵州医科大学药学院
• 2:贵州医科大学附属医院药学部

摘要(Abstract)：

目的 探究NPS-2143对胶质瘤U87、U251和Hs683细胞增殖与迁移能力的影响及机制。方法 将胶质瘤细胞分为3组，不加药物的对照组(Control组)、6μmol/L和8μmol/L NPS-2143处理组，通过细胞增殖实验CCK-8和细胞平板克隆形成实验检测不同浓度NPS-2143对U87和U251细胞活力与增殖的影响，通过流式细胞术实验检测不同浓度NPS-2143对细胞周期和凋亡的影响，Western blot检测细胞凋亡相关蛋白(Bax、Caspase 3)和细胞周期相关蛋白(P21、Cyclin D1)的表达水平；通过细胞划痕实验和细胞迁移实验检测NPS-2143对U87和Hs683细胞迁移的影响，Western blot法检测上皮间质转化(EMT)相关蛋白(N-cadherin、MMP2、MMP9)的表达水平；通过透射电镜观察NPS-2143对U87细胞中自噬囊泡数量的影响，Western blot检测细胞自噬相关蛋白(LC3、P62)和AKT-mTOR信号通路相关蛋白(p-AKT、p-mTOR)的表达水平。结果 与Control组比较，NPS-2143处理组胶质瘤细胞活力与增殖能力降低，且呈时间、浓度依赖性(P<0.01);与Control组比较，NPS-2143处理组胶质瘤细胞G1期细胞比例和凋亡细胞比例升高(P<0.05);与Control组比较，NPS-2143处理组胶质瘤细胞中P21蛋白表达水平增加，Cyclin D1蛋白表达水平下降，细胞凋亡蛋白Bax和Caspase 3表达水平增加(P<0.01);与Control组比较，NPS-2143抑制胶质瘤细胞的迁移能力(P<0.01),胶质瘤细胞中N-cadherin、MMP2、MMP9的表达水平降低(P<0.01),LC3、P62、p-AKT、p-mTOR蛋白表达水平升高(P<0.01);与Control组比较，NPS-2143使胶质瘤细胞内自噬囊泡增加(P<0.01)。结论 NPS-2143能抑制胶质瘤的增殖与迁移，其机制可能与激活AKT-mTOR通路有关。
Objective To explore the effect and mechanism of NPS-2143 on proliferation and migration of glioma U87 and U251, and Hs683 cells. Methods The glioma cells were divided into 3 groups: the Control group(without medical treatment), the 6 μmol/L and the 8 μmol/L NPS-2143 treatment group. The effects of NPS-2143 on the viability and proliferation of U87 and U251 cells were investigated by using cell counting kit-8(CCK-8) and colony formation experiments after the treatment with different concentrations of NPS-2143. Flow cytometer(FCM) was used to detect the effects of different concentrations of NPS-2143 on cycle and apoptosis of U87 and U251 cells. The expression levels of apoptosis-related proteins(Bax, Caspase 3) and cell cycle related proteins(P21, Cyclin D1) in U87 and U251 were detected by Western blot; The migration capability of U87 and Hs683 cells treated with different concentrations of NPS-2143 was detected by wound healing and Transwell experiments. The expression levels of epithelial-mesenchymal transition(EMT) related proteins(N-cadherin, MMP2, MMP9) in U87 and Hs683 cells were detected by Western blot; the expression levels of autophagy proteins(LC3, P62) and AKT-mTOR pathway related proteins(p-AKT, p-mTOR) in U87 and U251 cells were detected by Western blot. The changes of the number of autophagy vesicles in glioma U87 cells treated with NTP-2143 were observed by transmission electron microscopy. Results The viability and proliferation ability of glioma cells were reduced, with dependence of time and concentration(P<0.01), the proportaion of G1 phase and apoptotic glioma cells increased(P<0.05) and the expression level of P21, Bax, and Caspase 3 increased(P<0.01) while the level of Cyclin D1 decreased(P<0.01) in the NPS-2143 treatment group, compared with the Control group. NPS-2143 inhibited the migration ability of glioma cells(P<0.01) and decreased the expression levels of N-cadherin, MMP2, and MMP9(P<0.01), and also increased the expression levels of autophagy proteins LC3, P62, p-AKT, and p-mTOR(P<0.01), compared with the Control group. The number of autophagy vesicles obviously increased in glioma U87 cells treated with NPS-2143, compared with the Control group(P<0.01). Conclusion NPS-2143 may inhibit the proliferation and migration of glioma cells by blocking autophagy through mediating the AKT-mTOR pathway.

关键词(KeyWords)： 胶质瘤;NPS-2143;自噬;化疗;上皮间质转化
glioma;NPS-2143;autophagy;chemotherapy;epithelial-mesenchymal transition

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(82060775);; 贵州省科技项目(黔科合基础-ZK[2021]一般563)

作者(Author): 聂佳丽,殷海棠,文志鹏,郑志昌,李明,李琴,樊兴华,赵青青,张彦燕,杨继红
NIE Jiali,YIN Haitang,WEN Zhipeng,ZHENG Zhichang,LI Ming,LI Qin,FAN Xinghua,ZHAO Qingqing,ZHANG Yanyan,YANG Jihong

DOI: 10.19367/j.cnki.2096-8388.2024.02.001

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