摘要
目的:观察中药提取物姜黄素联合肿瘤坏死因子相关凋亡诱导配体对肺癌细胞株A549生长抑制率和对细胞凋亡相关因子表达的影响,探索姜黄素改善肺癌A549细胞对肿瘤坏死因子相关凋亡诱导配体抵抗的机制。方法:实验于2003-10/2004-10在上海东方医院中心实验室完成。用购于中科院上海细胞所肺腺癌细胞株A549,将培养的A549细胞暴露于姜黄素、肿瘤坏死因子相关凋亡诱导配体及两者联合中,分4组,每组6孔,姜黄素组(40μmol/L姜黄素)、肿瘤坏死因子相关凋亡诱导配体组(25μg/L肿瘤坏死因子相关凋亡诱导配体)、联合组(40μmol/L姜黄素与25μg/L肿瘤坏死因子相关凋亡诱导配体联合)、对照组(RPMI1640细胞培养液)。①用四氮唑蓝法测定姜黄素组、肿瘤坏死因子相关凋亡诱导配体组以及联合组的吸光值,根据吸光度值计算肺癌细胞株A549细胞生长抑制率。②应用ApoAlert半胱氨酸天冬氨酸蛋白酶系列比色法凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3,半胱氨酸天冬氨酸蛋白酶8表达。③反转录聚合酶链反应测定凋亡调控蛋白bcl-2/bax的表达。结果:①抑制率:肿瘤坏死因子相关凋亡诱导配体对肺腺癌A549细胞的抑制明显低于姜黄素,25μg/L肿瘤坏死因子相关凋亡诱导配体作用24h细胞抑制率仅有(2.65±1.416)%;两药联合对肺腺癌A549细胞的抑制作用明显增加[高达(30.97±1.318)%,P<0.01]。②凋亡相关因子:联合组半胱氨酸天冬氨酸蛋白酶3、半胱氨酸天冬氨酸蛋白酶8表达明显高于姜黄素组、肿瘤坏死因子相关凋亡诱导配体组、对照组[联合组:(50.77±0.812),(73.31±0.730)μmol/L;姜黄素组:(23.61±0.398),(26.84±1.511)μmol/L;肿瘤坏死因子相关凋亡诱导配体组:(9.63±1.024),(14.21±0.138)μmol/L;对照组:(3.69±0.486),(2.87±0.633)μmol/L,P<0.01];联合组、姜黄素组的bcl-2/bax明显高于肿瘤坏死因子相关凋亡诱导配体组(2.55±0.138,2.24±0.197,0.58±0.046,P<0.01)。结论:肺癌A549细胞对肿瘤坏死因子相关凋亡诱导配体细胞耐受,姜黄素可能通过调整bcl-2/bax的表达,进而激活半胱氨酸天冬氨酸蛋白酶家族,增加肿瘤坏死因子相关凋亡诱导配体的敏感性。
AIM： To observe the effect of the Chinese herb extract of curcumin combined with the tumor necrosis factor related apoptosis-inducing ligand （TRAIL） on the inhibitory rate of lung adenocarcinoma A549 cells and the expression of apoptosis-related factors, so as to explore the mechanism.of curcumin for enhancing the sensibility of lung cancer cells to TRAIL.
METHODS： Between October 2003 and October 2004, the experiment was carried out at the Central Laboratory of Shanghai East Hospital. The A549 cell bought from Shanghai Cell Institute of Chinese Academy of Sciences were cultured with curcumin, TRAIL and combination of curcumin and TRAIL, respectively and divided into 4 groups with 6 well in each group： curcumin group （40 μmol/L cureumin）, TRAIL group （25 μg/L TRAIL）, combination group （40 μmol/L eureumin and 25 μg/L TRAIL） and control group （RPMI1640 euhure fluid）. ①The absorbanee values of the eureumin group, TRAIL group and combination group were determined by MTT method to calculate the inhibitory rate of A549 cells. ②The expressions of easpase 8 and easpase 3 in each group were determined by ApoAlert Caspase Colorimetry set. ③The bel-2/bax expression of each group was measured by reverse transcription polymerase chain reaction. RESULTS： ①Inhibitory rate of A549 cells： The rate in the TRAIL group, which was （2.65±1.416）% euhured for 24 hours, was obviously lower than the eureumln group; whiIe it was remarkably increased by eombinatlon of TRAIL and eureumin [（30.97±1.318）%, P 〈 0.01]. ②The apoptosis-related factors： The expressions of easpase 3 and easpase 8 in the combination group were significantly higher than the other three groups [the combination group： （50.77 ±0.812）, （73.31 ±0.730） μmol/L; the curcumin group： （23.61 ±0.398）, （26.84 ± 1.511） μmol/L; the TRAIL group： （9.63 ± 1.024）, （14.21±0.138） μmol/L; the control group （3.69±0.486）, （2.87±0.633） μmol/L; P 〈 0.01]; The bcl-2/bax values of the combination and curcumin groups were remarkably higher than the TRAIL group （2.55±0.138, 2.24±0.197, 0.58±0.046, P 〈 0.01）.
CONCLUSION： Lung adenocarcinoma A549 cells are resistance to TRAIL; the curcumin can activate the caspase family to improve the sensibility of TRAIL by modulating the expression of bcl-2/bax.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第31期105-107,共3页
Chinese Journal of Clinical Rehabilitation
