ALCAT1缺失在KRAS诱导肺腺癌中的作用及机制研究  

The role and mechanism of ALCAT1 deletion in KRAS induced lung adenocarcinoma

作　　者：张潇月 郑月[1] 蒋厅 史裕光 ZHANG Xiaoyue;ZHENG Yue;JIANG Ting;SHI Yuguang(Department of Molecular Biology and Biochemistry,Nanjing Medical University,Nanjing 211166,China)

机构地区：[1]南京医科大学生物化学与分子生物学系,江苏南京211166

出　　处：《南京医科大学学报（自然科学版）》2022年第7期932-940,972,共10页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(31771309)。

摘　　要：目的:探究心磷脂酰基转移酶1(cardiolipin acyltransferase 1,ALCAT1)在Kirsten大鼠肉瘤病毒癌基因同源物(Kirsten rat sarcoma viral oncogene homolog,KRAS)诱导的肺腺癌中的作用及机制。方法:动物实验:在CC10rtta-KRAS小鼠中敲除ALCAT1基因,并使用多西环素饮食诱导肺腺癌发生,通过微计算机断层扫描(micro-computed tomography,micro-CT)以及HE染色分析小鼠肺腺癌的发生。细胞实验:将A549细胞分为对照组、Jenu组、缺氧组、缺氧+Jenu组,通过氯化钴(CoCl2)处理模拟缺氧条件,并使用ALCAT1抑制剂Jenu抑制其酶活性;试剂盒检测活性氧(reactive oxygen species,ROS)水平和乳酸水平,RT-PCR和Western blot分析相关信号分子的mRNA转录以及蛋白表达。结果:动物实验结果显示,ALCAT1缺失减缓小鼠肺腺癌发展,肺部肿瘤体积减小。细胞实验结果显示,Jenu显著降低缺氧条件下ROS和乳酸的生成,并下调缺氧诱导因子1α(hypoxia inducible factor 1α,HIF1α)表达以及Akt磷酸化水平。结论:ALCAT1缺失可以抑制ROS生成并下调HIF1α表达以及Akt磷酸化水平,从而改善肿瘤糖酵解代谢,最终减缓肺腺癌的发展。Objective:To investigate the role and mechanism of cardiolipin acyltransferase 1(ALCAT1)in Kirsten rat sarcoma viral oncogene homolog(KRAS)-induced lung adenocarcinoma.Methods:In the animal experiments,the CC10rtta-KRAS mice were knocked out of the ALCAT1 gene,and induced lung adenocarcinoma with doxycycline diet.The occurrence of lung adenocarcinoma in mice was analyzed by micro-computed tomography(micro-CT)and HE staining.In the cell experiments,A549 cells were divided into the control group,Jenu group,hypoxia group and hypoxia+Jenu group,cobalt chloride(CoCl2)treatment was used to simulate hypoxic conditions,and ALCAT1 inhibitor Jenu was used to inhibit its enzymatic activity.The reactive oxygen species(ROS)and lactate levels were measured using kits in cells.Moreover,the mRNA transcription and protein expression of the related signal molecules were detected by RT-PCR and Western blot.Results:The results of animal experiments showed that ALCAT1 deficiency slowed the development of lung adenocarcinoma in mice and reduced lung tumor size.The results of cell experiments showed that Jenu significantly decreased the production of ROS and lactate under hypoxia,and down-regulated the expression of hypoxia inducible factor 1α(HIF1α)and the phosphorylation level of Akt.Conclusion:ALCAT1 deficiency could inhibit ROS generation and down-regulate HIF1αexpression and Akt phosphorylation level,thereby improving the glycolysis metabolism in the tumor,ultimately preventing the development of lung adenocarcinoma.

关 键 词：心磷脂酰基转移酶1 肺腺癌 活性氧 缺氧诱导因子1Α 

分 类 号：R734.2[医药卫生—肿瘤]