CD147通过Akt/mTOR信号通路调控脂肪酸合成对子宫颈癌细胞增殖、侵袭和迁移的影响

CD147 regulates the effects of fatty acid synthesis on the proliferation,invasion,and migration of cervical cancer cells via the Akt/mTOR signaling pathway

目的探讨CD147的表达对子宫颈癌细胞增殖、侵袭、迁移的影响和潜在的分子机制。方法分析UCSC数据库BSG基因(编码CD147蛋白)在子宫颈癌中的表达,采用Log-rank test法评估各组表达的预后差异。应用Western blot法检测CD147在Siha、Hela和H8细胞中的表达,通过慢病毒转染Hela细胞下调CD147表达,并验证其转染效率。运用Western blot法检测各组细胞中p-Akt、p-mTOR、ACC1、FASN、E-cadherin和N-cadherin的表达;使用BODIPY染色和脂肪酸试剂盒检测细胞中脂肪酸含量;利用CCK-8、平板克隆和Transwell实验检测细胞增殖、侵袭和迁移能力。sh-CD147组经Akt激动剂(SC79)处理后,Western blot法检测细胞中p-Akt、p-mTOR、ACC1、FASN、E-cadherin和N-cadherin的表达;选用BODIPY染色和脂肪酸试剂盒检测细胞中脂肪酸含量;利用平板克隆实验和Transwell实验分别检测细胞增殖、侵袭和迁移能力。结果UCSC数据库显示CD147在子宫颈癌中的表达高于正常子宫颈组织(P<0.01);CD147过表达的患者预后不良。Western blot检测结果显示,与H8细胞相比,Siha和Hela细胞中CD147蛋白表达增高(P=0.011)。下调CD147表达后,与Hela组相比,sh-CD147组细胞中CD147、ACC1和FASN蛋白表达降低(P<0.001);BODIPY荧光染色减弱,脂肪酸含量下降(P<0.001);细胞集落形成能力、侵袭和迁移能力降低;sh-CD147组细胞中E-cadherin蛋白表达升高,N-cadherin、p-Akt和p-mTOR表达降低。与sh-CD147相比,经Akt激动剂SC-79剂处理后(sh-CD147-SC79)细胞内p-Akt、p-mTOR、ACC1、FASN和N-cadherin表达升高,E-cadherin表达降低,脂质染色和脂肪酸含量检测结果与关键酶表达一致(P<0.01),细胞增殖、侵袭迁移能力显著增强。结论CD147通过Akt/mTOR信号通路调控脂肪酸合成,促进子宫颈癌细胞增殖、侵袭和迁移。

Purpose To investigate the effect of CD147 on the proliferation,invasion and migration of cervical cancer cells and its potential molecular mechanism.Methods The expression data of BSG gene(encoding CD147 protein)in cervical cancer samples were downloaded from UCSC database,and the prognosis of different groups of samples was evaluated by Log-rank test.Western blot was used to detect CD147 expression in the Siha and Hela and H8 cells.The expression of CD147 was downregulated by the lentivirus transfection into Hela cells and its transfection efficiency was verified.Western blot was used to detect the expression of p-Akt,p-mTOR,ACC1,FASN,E-cadherin and N-cadherin in each group.The content of fatty acids in the cells was detected by BODIPY staining and fatty acid kit.Cell proliferation,invasion and migration were detected by CCK-8,plate cloning and Transwell assay.The cell proliferation,invasion and migration ability were detected by plate cloning experiment and Transwell test.Results The expression of CD147 in cervical cancer tissues was higher than that in normal cervical tissues(P<0.01).Patients with overexpression of CD147 had poor prognosis.Western blot results showed that compared with H8 cells,the expression of CD147 protein in Siha and Hela cells was increased(P=0.011).After down-regulation of CD147,the protein expression of CD147,ACC1 and FASN in the sh-CD147 group was decreased compared with those in the Hela group(P<0.001).BODIPY fluorescence staining was weakened and fatty acid content was decreased(P<0.001).The ability of cell colony formation,invasion and migration was decreased.The expression of E-cadherin protein in sh-CD147 group was increased,and the expression of N-cadherin,p-Akt and p-mTOR was decreased.Compared to sh-CD147,after treatment with Akt agonist SC-79(sh-CD147-SC79),the expression of p-Akt,p-mTOR,ACC1,FASN,N-cadherin in cells was increased,and the expression of E-cadherin was decreased,and the results of lipid staining and fatty acid content were consistent with the expression of key enzymes(P<0.01),and the cell proliferation,invasion and migration ability were significantly enhanced.Conclusion CD147 through Akt/mTOR signaling pathways regulating the fatty acid synthesis promotes cervical cancer cell proliferation,invasion and migration.