CAJ | 학술논문

目的：研究三氧化二砷( arsenic trioxide, As2 O3)对腺样囊性癌( adenoid cystic carcinoma,ACC)中ACC-2细胞增殖、凋亡的影响,并从基因和蛋白水平分析其对ACC-2细胞中MDM2基因表达的影响,从而探讨As2 O3诱导ACC-2细胞凋亡的具体机制。 　　方法：体外培养ACC-2细胞,分为实验组和对照组,向培养至指数生长期的 ACC-2细胞加入不同浓度(2、4、6、8μmol/L)的含As2 O3的细胞培养液作为实验组,对照组给予等量的细胞培养液,加入 As2 O3后分别培养不同时间。倒置显微镜下密切观察各时间点不同浓度As2 O3对ACC-2细胞的生长抑制作用和细胞凋亡的形态变化,应用荧光定量RT-PCR和免疫细胞化学技术检测MDM2基因(24、48h)和蛋白(24、48、72h)的表达变化。 　　结果：经As2 O3处理后ACC-2细胞体积缩小变圆,核质固缩,悬浮凋亡细胞增多,存活细胞数量明显减少。 RT-PCR及免疫细胞化学结果一致显示, ACC-2细胞随着药物浓度的增加及作用时间的延长,MDM2表达明显减少,与对照组比较,差异有统计学意义(P<0.05),不同浓度不同时间各组两两比较差异有统计学意义(P<0.05),MDM2蛋白表达量与浓度及时间呈显著负相关性( r<-0.7,P<0.05),即呈浓度和时间依赖性。 　　结论：As2 O3对ACC-2细胞具有生长抑制和诱导凋亡的作用,且As2 O3能够下调腺样囊性癌ACC-2细胞中癌基因MDM2 mRNA及蛋白的表达,可能是其诱导腺样囊性癌ACC-2细胞凋亡的机制。
목적：연구삼양화이신( arsenic trioxide, As2 O3)대선양낭성암( adenoid cystic carcinoma,ACC)중ACC-2세포증식、조망적영향,병종기인화단백수평분석기대ACC-2세포중MDM2기인표체적영향,종이탐토As2 O3유도ACC-2세포조망적구체궤제。 　　방법：체외배양ACC-2세포,분위실험조화대조조,향배양지지수생장기적 ACC-2세포가입불동농도(2、4、6、8μmol/L)적함As2 O3적세포배양액작위실험조,대조조급여등량적세포배양액,가입 As2 O3후분별배양불동시간。도치현미경하밀절관찰각시간점불동농도As2 O3대ACC-2세포적생장억제작용화세포조망적형태변화,응용형광정량RT-PCR화면역세포화학기술검측MDM2기인(24、48h)화단백(24、48、72h)적표체변화。 　　결과：경As2 O3처리후ACC-2세포체적축소변원,핵질고축,현부조망세포증다,존활세포수량명현감소。 RT-PCR급면역세포화학결과일치현시, ACC-2세포수착약물농도적증가급작용시간적연장,MDM2표체명현감소,여대조조비교,차이유통계학의의(P<0.05),불동농도불동시간각조량량비교차이유통계학의의(P<0.05),MDM2단백표체량여농도급시간정현저부상관성( r<-0.7,P<0.05),즉정농도화시간의뢰성。 　　결론：As2 O3대ACC-2세포구유생장억제화유도조망적작용,차As2 O3능구하조선양낭성암ACC-2세포중암기인MDM2 mRNA급단백적표체,가능시기유도선양낭성암ACC-2세포조망적궤제。
AIM:To investigate effects of arsenic trioxide ( As2 O3 ) on the proliferation and apoptosis of on adenoid cystic carcinoma-2 ( ACC-2 ) cells and detect the expression of MDM2 gene from gene and protein level and to explore detailed mechanism of As2 O3 inducing ACC - 2 cells apoptosis. METHODS: ACC-2 cells were cultured in vitro and divided into the experiment group and control group. Different concentrations of As2 O3(2, 4, 6, 8μmol/L) were applied to cells in logarithmic growth phase at different time as experiment group, the control group was given the same amount of cell culture fluid, after added As2 O3 , the cells were cultured at different times, respectively. The effect of different As2 O3 concentrations at each point time on inhibition and metamorphoses of ACC-2 cells was observed under inverted phase contrast microscope. Expression changes of MDM2 mRNA ( 24, 48h ) and protein ( 24, 48, 72h ) were determined by reverse transcription polymerase chain reaction ( RT-PCR ) and immunohistochemistry test ( IHCT) respectively. RESULTS: Cells shrinkage, nuclear chromatin condensation, apoptotic cells increased and the number of viable cells significantly reduced after being cultured with different concentrations of As2 O3 . The results of RT-PCR and IHCT were showed consistent the expression of MDM2 in experiment group decreased gradually with the increase of As2 O3 concentrations and extension of action time, which was significantly different to that in the control group (P<0. 05). Campared with each other, it was statistically significant between the different concentration and time of two groups (P<0. 05). MDM2 expression was negatively correlated with concentration and time (r<-0. 7, P<0. 05), that was, it presented in dose- and time-dependent manner. CONCLUSION:As2 O3 has the inhibitory and apoptosis-inducing effect on ACC-2 cells, and it can downregulate the expression of MDM2 mRNA and protein in ACC-2 cell line. This may be the mechanism of As2 O3 induced ACC-2 cells apoptosis.