CAJ | 학술논문

目的:探讨芹菜素(Apigenin)对乳腺癌MDA-MB-231细胞系凋亡的影响.方法:常规培养MDA-MB-231细胞,应用四甲基偶氮唑盐(MTT)法评价芹菜素对乳腺癌MDA-MB-231细胞增殖的影响.荧光染色法观察细胞形态学变化.流式细胞术(FCM)Annexin V/PI 双染法检测细胞凋亡率.Western blot检测不同浓度芹菜素对Caspase-3,PARP,突变型P53,P73,PIG3,Bax, Bcl-2等凋亡相关蛋白表达的影响.RNA干扰技术检测P73对凋亡及下游靶基因PIG3表达的影响.结果:MTT结果显示,芹菜素10、20及40μM分别处理细胞24 h,发现芹菜素可有效抑制MDA-MB-231细胞增殖,且具有浓度依赖性和时间依赖性,实验组与对照组比较差异具有统计学意义(P<0.05),两组组间比较差异具有统计学意义(P<0.05);荧光染色结果显示,对照组未观察到凋亡细胞,芹菜素10μM、20μM及40μM组细胞中均可见凋亡小体,其数目随芹菜素浓度的增加而逐渐增加,具有浓度依赖性;流式细胞术结果显示,与对照组比较,芹菜素10μM,20μM及40μM组早期凋亡率逐渐增加,呈明显的量效关系,实验组与对照组比较差异具有统计学意义(P<0.05);Western blot结果显示,与对照组相比,芹菜素10μM、20μM及40μM组Caspase-3、PARP剪切带含量明显增加,P73、PIG3及Bax表达增加,而突变型P53与Bcl-2表达减少,且具有浓度依赖性.RNA干扰结果显示,与阴性对照组相比,p73-siRNA 可有效抑制芹菜素诱导的PIG3表达,Caspase-3和PARP剪切含量也相应降低,实验组与对照组比较差异具有统计学意义(P<0.05).结论:芹菜素可通过非P53依赖性凋亡有效抑制乳腺癌MDA-MB-231细胞增殖,其机制可能与下调突变型p53表达,上调P73介导的PIG3表达,以及Bax/Bcl-2比值增加有关.
목적:탐토근채소(Apigenin)대유선암MDA-MB-231세포계조망적영향.방법:상규배양MDA-MB-231세포,응용사갑기우담서염(MTT)법평개근채소대유선암MDA-MB-231세포증식적영향.형광염색법관찰세포형태학변화.류식세포술(FCM)Annexin V/PI 쌍염법검측세포조망솔.Western blot검측불동농도근채소대Caspase-3,PARP,돌변형P53,P73,PIG3,Bax, Bcl-2등조망상관단백표체적영향.RNA간우기술검측P73대조망급하유파기인PIG3표체적영향.결과:MTT결과현시,근채소10、20급40μM분별처리세포24 h,발현근채소가유효억제MDA-MB-231세포증식,차구유농도의뢰성화시간의뢰성,실험조여대조조비교차이구유통계학의의(P<0.05),량조조간비교차이구유통계학의의(P<0.05);형광염색결과현시,대조조미관찰도조망세포,근채소10μM、20μM급40μM조세포중균가견조망소체,기수목수근채소농도적증가이축점증가,구유농도의뢰성;류식세포술결과현시,여대조조비교,근채소10μM,20μM급40μM조조기조망솔축점증가,정명현적량효관계,실험조여대조조비교차이구유통계학의의(P<0.05);Western blot결과현시,여대조조상비,근채소10μM、20μM급40μM조Caspase-3、PARP전절대함량명현증가,P73、PIG3급Bax표체증가,이돌변형P53여Bcl-2표체감소,차구유농도의뢰성.RNA간우결과현시,여음성대조조상비,p73-siRNA 가유효억제근채소유도적PIG3표체,Caspase-3화PARP전절함량야상응강저,실험조여대조조비교차이구유통계학의의(P<0.05).결론:근채소가통과비P53의뢰성조망유효억제유선암MDA-MB-231세포증식,기궤제가능여하조돌변형p53표체,상조P73개도적PIG3표체,이급Bax/Bcl-2비치증가유관.
Objective: This work aims to investigate the anticancer effect of apigenin in the MDA-MB-231 breast cancer cell line. Methods: The MDA-MB-231 cells were cultured in vitro. Cell proliferation was measured using the methyl thiazolyl tetrazolium (MTT) assay. Morphological changes in the apoptotic cells were observed by fluorescent microscopy. Flow cytometry was used to detect the rate of apoptotic cells with longer treatments of apigenin. Apoptosis-related proteins were detected by Western blot. The ribonucleic acid in-terference (RNAi) experiment was applied using chemically synthesized siRNA. Results: The MTT assay revealed that cell proliferation significantly decreased in a dose- and time-dependent manner. Fluorescent staining and flow cytometry showed that the number of apoptotic cells increased with a longer treatment of apigenin. Caspase 3 activation and the poly ADP ribose polymerase (PARP) cleavage were observed after treatment with apigenin, indicating the death of apoptotic cells. The upregulation of p73, PIG3, and Bax expressions, as well as the downregulation of mutant p53 and Bcl-2 expressions, was also observed in the apigenin-treated cells. The RNAi assay showed that the silencing of p73 significantly inhibited PIG3 expression, as well as the cleavage of Caspase 3 and PARP induced by the apigenin treatment. Conclusion: Apigenin exhibits an inhibitory effect against cell proliferation by inducing the p53-independent apoptosis of the MDA-MB-231 cells. The underlying mechanism may be associated with the downregulation of mutant p53, the upregulation of p73-mediated PIG3, and the increase of the Bax/Bcl-2 ratio.