乌骨藤总苷对肺腺癌NCI-H1299细胞糖酵解、凋亡和自噬作用的影响  

Effect of total glycosides in Glaucescent Fissistigma root on glycolysis,apoptosis and autophagy of lung adenocarcinoma NCI-H1299 cells

作　　者：王晨亮 张菁 刘浩茹 李观华 WANG Chenliang;ZHANG Jing;LIU Haoru;LI Guanhua(Departmen of Pathology,Jiujiang No.1 People′s Hospital,Jiangxi Province,Jiujiang 332000,China)

机构地区：[1]不详

出　　处：《中国当代医药》2022年第27期15-19,24,共6页China Modern Medicine

基　　金：江西省卫生健康委科技计划项目(SKJP220201685)。

摘　　要：目的探讨乌骨藤总苷通过Kirsten大鼠肉瘤病毒癌基因同源物(KRAS)对肺腺癌细胞糖酵解、凋亡和自噬的影响。方法体外培养NCI-H1299细胞,分为对照组、乌骨藤总苷低剂量组、乌骨藤总苷高剂量组、乌骨藤总苷+Vector组和乌骨藤总苷+KRAS组。对照组用正常培养基作用于细胞,乌骨藤低剂量组用终浓度为5 mg/ml的乌骨藤总苷作用于细胞,乌骨藤高剂量组用终浓度为20 mg/ml的乌骨藤总苷作用于细胞,乌骨藤总苷+Vector组先在细胞内转染入阴性对照载体,再用20 mg/ml的乌骨藤总苷作用于细胞,乌骨藤总苷+KRAS组转染入KRAS过表达载体后,用20 mg/ml的乌骨藤总苷作用于细胞。用qRT-PCR法检测各组细胞中KRAS mRNA的表达情况,用Western blot法检测各组细胞中KRAS蛋白的表达情况,用MTT法检测细胞增殖抑制率,用试剂盒检测各组细胞葡萄糖消耗量和乳酸含量,用流式细胞术检测细胞凋亡率,用Western blot法检测各组细胞中凋亡和自噬相关蛋白的表达水平。结果乌骨藤总苷低剂量组和乌骨藤总苷高剂量组KRAS表达水平、葡萄糖消耗量、乳酸含量、Bcl-2蛋白水平低于对照组,细胞增殖抑制率和凋亡率、LC3-Ⅱ/Ⅰ、beclin-1蛋白和Bax蛋白水平高于对照组,差异均有统计学意义(P<0.05)。乌骨藤总苷+KRAS组细胞中KRAS水平、葡萄糖消耗量和乳酸含量、Bcl-2蛋白表达水平均高于乌骨藤总苷+Vector组,差异有统计学意义(P<0.05)。乌骨藤总苷+KRAS组细胞增殖抑制、细胞凋亡率、LC3-Ⅱ/Ⅰ、Bax蛋白和beclin-1蛋白水平低于乌骨藤总苷+Vector组,差异有统计学意义(P<0.05)。乌骨藤总苷高剂量组和乌骨藤总苷+Vector的各项指标比较,差异均无统计学意义(P>0.05)。结论乌骨藤总苷可以通过抑制KRAS的表达,抑制肺腺癌细胞的糖酵解过程,促进肺腺癌细胞凋亡和自噬,从而对肺腺癌细胞起到抑制作用。Objective To explore the effect of total glycosides in Glaucescent Fissistigma root on glycolysis,apoptosis and autophagy of lung adenocarcinoma cells through Kirsten rats arcoma viral oncogene homolog(KRAS).Methods The NCI-H1299 cells were cultured in vitro and divided into the control group,the low-dose group of total glycosides in Glaucescent Fissistigma root,the high-dose group of total glycosides in Glaucescent Fissistigma root,the group of total glycosides in Glaucescent Fissistigma root+Vector,and the group of total glycosides in Glaucescent Fissistigma root+KRAS.The control group was treated with normal cultue medium,while the low-dose group received final concentration of 5 mg/ml of total glycosides in Glaucescent Fissistigma root,the high-dose group received final concentration of 20 mg/ml of total glycosides in Glaucescent Fissistigma root.The cells in the total glycosides in Glaucescent Fissistigma root+Vector group were transfected with a negative control vector first,and then the cells were treated with 20 mg/ml of total glycosides in Glaucescent Fissistigma root.After the cells were transfected with a KRAS overexpression vector,the total glycosides in Glaucescent Fissistigma root+KRAS group were treated with 20 mg/ml of total glycosides in Glaucescent Fissistigma root.The expression of KRAS mRNA in cells of each group was detected by qRT-PCR,and the expression of KRAS protein in cells of each group was detected by Western blot.And the cell proliferation inhibition rate was detected by MTT method.The glucose consumption and lactic acid content of cells in each group were detected by kit.The apoptosis rate was detected by flow cytometry,and the expression levels of apoptotic and autophagy-related proteins in cells of each group were determined by Western blot. Results The expression levels of KRAS, glucose consumption, lactic acid content, and Bcl-2 protein in the low-dose group and the high-dose group were lower than those in the control group, the cell proliferation inhibition rate and apoptosis ra

关 键 词：乌骨藤总苷 Kirsten大鼠肉瘤病毒癌基因同源物 糖酵解 凋亡和自噬 肺腺癌细胞 

分 类 号：R285.5[医药卫生—中药学]