康莱特注射液对人肺腺癌细胞A549凋亡的影响及机制

Effect of Kanglaite injection on cell apoptosis of human pneumonic adenocarcinoma cell line A549 and its mechanism

目的观察薏苡仁注射液(康莱特注射液)对人肺腺癌细胞A594凋亡的影响,并探讨康莱特抗肺腺癌细胞的作用及机制。方法体外培养人肺腺癌细胞株A549,分别加入5,10,15,20,25 mL/L康莱特注射液,对照组加等量生理盐水,孵育48 h后,采用流式细胞术(FCM)测定细胞周期的变化;20 mL/L康莱特注射液孵育48 h后,收集细胞,FCM测定细胞凋亡率,逆转录-聚合酶链反应(RT-PCR)法、免疫印迹(Western blot)法分别测定Caspase-3、Bcl-2、Bax、Fas、FasL mRNA和蛋白的表达情况。结果康莱特注射液处理后,G1期的细胞均显著增加,S期细胞明显减少;20 mL/L康莱特注射液处理A549细胞48 h后,细胞凋亡率、Caspase-3、Bax、Fas、FasL mRNA和蛋白表达显著升高,Bcl-2 mRNA和蛋白表达水平下降,Bcl-2/Bax比值显著降低。结论康莱特注射液可通过上调A549细胞Caspase-3、Fas和FasL的表达,降低Bcl-2/Bax比值,诱导其凋亡,产生抗肿瘤作用。

ObjectiveIt is to observe the effects of Kanglaite injection on cell apoptosis of human pneumonic adenocarcinoma cell line A549,and further to explore its anti-adenocarcinoma function and the mechanisms.Methods A549 cells were cultured in vitro,and were cultured with Kanglaite（5,10,15,20,25 mL/L） for 48 h,the cells in the control group were cultured with the same dosage of normal saline.Then the change of cell cycles was detected by flow cytometry（FCM）.Cells treated with 20 mL/L Kanglaite for 48 h were collected to determine the expression of Caspase-3,Bcl-2,Bax,Fas,FasL mRNA by RT-PCR and Western blot assays,And the cell apoptosis rate was obtained by FCM.Results After A549 cells were treated with Kanglaite,the cell number of G1 phase was increased,while the cell number of S phase was decreased.After the cells cultured with 20 mL/L kanglaite injection for 48 h,the cell apoptosis rate and the expression of Caspase-3,Bax,Fas,FasL mRNA obviously increased,and the expression of Bcl-2 and Bcl-2/Bax decreased significantly.ConclusionKanglaite injection can induce cell apoptosis by up-regulating the caspase-3,Fas and FasL expression and decreasing the rate of Bcl-2/Bax,so to pay a anti-tumor function.