摘要
目的探讨磷脂酶Cε1(PLCE1)表达与肺腺癌A549细胞凋亡的关系及作用机制。方法将肺腺癌细胞株A549分为空白组(仅给予正常培养液)、实验A组(正常培养液+5mmol/L PLCE1抑制剂U73122共培养)、实验B组(正常培养液+10mmol/L PLCE1抑制剂U73122共培养),观察各组,采用流式细胞仪技术检测培养12h、24h及48h时刻A549细胞的凋亡率,采用RT-PCR及Western-blot技术检测各组共培养48h后的PLCE1、p53、Bcl-2、Bax mRNA及蛋白表达水平。结果在培养12h、24h、48h后,实验A组、实验B组的肺腺癌A549细胞凋亡率显著高于空白组(P<0.05);实验B组的肺腺癌A549细胞凋亡率显著高于实验A组(P<0.05);在培养48h后,实验A组、实验B组的p53、Bax mRNA及蛋白水平显著高于空白组(P<0.05),实验A组、实验B组的Bcl-2、PLCE1 mRNA及蛋白水平显著低于空白组(P<0.05);在培养48h后,实验B组的p53、Bax mRNA及蛋白水平显著高于实验A组(P<0.05),实验B组的Bcl-2、PLCE1 mRNA水平显著低于实验A组(P<0.05)。结论通过抑制肺腺癌A549细胞PLCE1的表达,进而抑制上调细胞中p53、Bax表达,下调Bcl-2表达,从而达到促进肺腺癌A549细胞凋亡的目的。
Objective To investigate the relationship between the expression of phospholipase C epsilon 1(PLCE1)and apoptosis of lung adenocarcinoma A549 cells and its mechanism.Methods The lung adenocarcinoma A549 cells were divided into blank group(only given normal medium),experimental group A(normal medium +5 mmol/L PLCE1 inhibitor U73122 co culture),experimental group B(normal medium +10 mmol/L PLCE1 inhibitor U73122 co culture).Apoptosis rate of cultured A549 cells was observed on 12 h,24 h and 48 h using flow cytometry;and on 48 h co cultured,PLCE1,p53,Bcl-2,Bax and mRNA protein expression were detected by RT-PCR and Western-blot technology.Results On 12 h,24 h and 48 h,apoptosis rate of lung adenocarcinoma A549 cells in culture was higher than the control group in the experimental group A and experimental group B(P<0.05);and it was significantly higher in the experimental group B than the experimental group A(P<0.05).After 48 h in culture,p53,Bax mRNA and protein level of the experimental group A and experimental group B was significantly higher than that of the control group(P<0.05),but their Bcl-2,PLCE1 mRNA and protein level was significantly lower than that of the control group(P<0.05);and p53,Bax mRNA and protein level of the experimental group B was also significantly lower than the experimental group A(P<0.05).Conclusion By inhibiting the expression of PLCE1 in lung adenocarcinoma A549 cells,the up regulation of p53 and Bax expression and down regulating the expression of Bcl-2 will be inhibited,so as to promote the apoptosis of lung adenocarcinoma A549 cells.
作者
彭盘俐
吕俊宏
林少欢
蔡长青
Peng Panli;Lv Junhong;Lin Shaohuan(The Second Department of Tumor,The Second Provincial General Hospital of Guandong ,Guangzhou510037,China)
出处
《中国煤炭工业医学杂志》
2018年第6期604-607,共4页
Chinese Journal of Coal Industry Medicine
基金
广东省医学科研基金项目(编号:B2013062)
