CAJ | 학술논문

目的：探讨康莱特注射液联合吉非替尼对人肺腺癌A549细胞株增殖和凋亡的影响。方法应用MTT法计算康莱特注射液作用于人肺腺癌A549细胞株48 h的半数抑制浓度（ IC50），检测康莱特注射液（5、10、20、40、80、100、160μl／ml）、吉非替尼（0?1、0?5、1?0、5、10、20、40μmol／L）以及康莱特注射液（80μl／ml）联合吉非替尼（0?1、0?5、1、5、10、20、40μmol／L）作用于人肺腺癌A549细胞株24、48、72 h的增殖抑制率；流式细胞仪Annexin V／PI 双标法检测康莱特注射液、吉非替尼及两药联合作用于A549细胞48 h后的凋亡率；免疫细胞化学法检测各药物处理组作用于A549细胞48 h后FAS、AKT2蛋白表达；RT?PCR技术检测各组FAS mRNA、AKT2 mRNA的表达水平。均设未加药的A549细胞为空白对照组。结果康莱特注射液作用于人肺腺癌A549细胞株48 h的IC50为80μl／ml。康莱特注射液联合吉非替尼作用于A549细胞的增殖抑制率高于两单药组和空白对照组，且呈时间和浓度依赖性（ P＜0?05）。两者的联合作用在一定浓度范围内呈现出协同的抗瘤效果。流式细胞术检测显示，两药联合组的细胞凋亡率显著高于两单药组和空白对照组（ P＜0?05）。免疫细胞化学和RT?PCR检测显示，与空白对照组比较，康莱特组和两药联合组FAS蛋白和mRNA的表达水平均升高（ P＜0?05），AKT2蛋白和mRNA表达水平均下调（ P＜0?05）。结论康莱特注射液联合吉非替尼对人肺腺癌A549细胞株有明显的促凋亡作用，康莱特注射液可能增加人肺腺癌A549细胞对于吉非替尼的敏感性。
목적：탐토강래특주사액연합길비체니대인폐선암A549세포주증식화조망적영향。방법응용MTT법계산강래특주사액작용우인폐선암A549세포주48 h적반수억제농도（ IC50），검측강래특주사액（5、10、20、40、80、100、160μl／ml）、길비체니（0?1、0?5、1?0、5、10、20、40μmol／L）이급강래특주사액（80μl／ml）연합길비체니（0?1、0?5、1、5、10、20、40μmol／L）작용우인폐선암A549세포주24、48、72 h적증식억제솔；류식세포의Annexin V／PI 쌍표법검측강래특주사액、길비체니급량약연합작용우A549세포48 h후적조망솔；면역세포화학법검측각약물처리조작용우A549세포48 h후FAS、AKT2단백표체；RT?PCR기술검측각조FAS mRNA、AKT2 mRNA적표체수평。균설미가약적A549세포위공백대조조。결과강래특주사액작용우인폐선암A549세포주48 h적IC50위80μl／ml。강래특주사액연합길비체니작용우A549세포적증식억제솔고우량단약조화공백대조조，차정시간화농도의뢰성（ P＜0?05）。량자적연합작용재일정농도범위내정현출협동적항류효과。류식세포술검측현시，량약연합조적세포조망솔현저고우량단약조화공백대조조（ P＜0?05）。면역세포화학화RT?PCR검측현시，여공백대조조비교，강래특조화량약연합조FAS단백화mRNA적표체수평균승고（ P＜0?05），AKT2단백화mRNA표체수평균하조（ P＜0?05）。결론강래특주사액연합길비체니대인폐선암A549세포주유명현적촉조망작용，강래특주사액가능증가인폐선암A549세포대우길비체니적민감성。
Objective To investigate the effects of Kanglaite injection( KLT) combined with gefitinib on the lung adenocarci?noma A549 cell proliferation and apoptosis in vitro. Methods MTT assay was applied to calculate the half inhibitory concerntration ( IC50 ) of KLT acting on human lung adenocarcinoma A549 cells for 48 h, and detect the inhibition rates by treating A549 cells with KLT(5,10,20,40,80,100,160 μl/ml), gefitinib(0?1,0?5,1?0,5,10,20,40μmol/L) and KLT(80μl/ml) combined with different doses of gefitinib for 24,48,72 h. Flow cytometry( FCM) was employed to detect the cell apoptosis for 48 h. Immunocytochemical and RT?PCR methods were used to detect the protien and mRNA express of AKT2 and FAS,respectively. Cells without drug was set as blank control group. Results MTT assay showed that IC50 of KLT acting on A549 cells for 48 h was 80 μl/ml. Compared with both single drug groups and control group, KLT combined with gefitinib could significantly inhibit the proliferation of A549 cells in a time?and dose?dependent manner. FCM showed that the cell apoptotic rates of combined group higher than those of both single drug groups and control group( P<0?05) . Immunocytochemical and RT?PCR detection showed that compared with the control group, the FAS pro?tein and mRNA levels of the KLT in combined group were increased, but AKT2 decreased( P<0?05) . Conclusion The combination of KLT and gefitinib can induce cell apoptosis significantly. KLT may increase the sensitivity of the human lung adenocarcinoma cell line A549 for gefitinib.