阳和汤对人乳腺癌MDA-MB-231细胞凋亡的影响
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摘要
目的:探讨阳和汤对人乳腺癌MDA-MB-231细胞凋亡的影响及其作用机制。方法:CCK-8检测阳和汤提取物不同浓度(0、10、20、40、80、120μg/ml)分别在24小时、48小时、72小时对MDA-MB-231细胞增殖的影响。设空白对照组(给等量培养基),阳和汤低、中、高剂量组(分别按10、20、40μg/ml浓度给药),药物干预24小时后,流式细胞术检测各组细胞凋亡的情况;免疫蛋白印迹法检测各组细胞Bcl-2、Bax、p-Akt、total Akt、pPI3K、total PI3K表达情况。结果:相对于空白对照组,阳和汤各剂量组细胞增殖的抑制率均增高,且对细胞增殖的抑制作用呈时间与剂量依赖,差异有统计学意义(P<0. 05)。药物干预24小时后,相对于空白对照组,阳和汤各剂量组MDA-MB-231细胞凋亡率均增加,差异具有统计学意义(P<0. 05); Bax表达增加、Bcl-2表达降低,p-PI3K相对PI3K、p-Akt相对Akt表达均降低,差异有统计学意义(P<0. 05)。结论:阳和汤可抑制人乳腺癌MDA-MB-231细胞的增殖、诱导其凋亡,其作用机制可能与其抑制PI3K/Akt信号通路的活化有关。
Objiective: To explore the effect and mechanism of Yanghe Decoction on apoptosis of MDA-MB-231 cells for human breast cancer. Methods: CCK-8 was used to detect the proliferation of MDA-MB-231 cells at 24 h,48 h and 72 h respectively at different concentrations( 0,10,20,40,80,120μg/ml). The controlled group( the same amount of culture medium),Yanghe Decoction low,medium and high dose groups( 10,20,40μg/ml) were established. The groups were intervened by the drug 24 h,and the flow cytometry was used to detect the apoptosis rate. The expression of Bcl-2,Bax,p-Akt,total Akt,p-PI3 K and total PI3 K were detected by Western blotting. Results: Compared with the controlled group,the inhibitory rate of cell proliferation was increased in different dose groups of Yanghe Decoction,and the inhibitory effect on cell proliferation was time and dose dependent( P < 0. 05). After 24 h of drug intervention,the apoptotic rate of MDA-MB-231 cells in each dose group was increased compared with that of the controlled group,the difference was statistically significant( P < 0. 05). Compared with the normal controlled group,the expression of Bax and Bcl-2 were decreased,and the expression of p-PI3 K was decreased relative to PI3 K,p-Aktwas decreasedrelative to Akt,and the difference was statistically significant( P < 0. 05). Conclusions: Yanghe Decoction can inhibit the proliferation and induce apoptosis of human breast cancer MDA-MB-231 cells,and its mechanism may be related to its inhibition of activation of PI3 K/Akt signaling pathway.
Objiective: To explore the effect and mechanism of Yanghe Decoction on apoptosis of MDA-MB-231 cells for human breast cancer. Methods: CCK-8 was used to detect the proliferation of MDA-MB-231 cells at 24 h,48 h and 72 h respectively at different concentrations( 0,10,20,40,80,120μg/ml). The controlled group( the same amount of culture medium),Yanghe Decoction low,medium and high dose groups( 10,20,40μg/ml) were established. The groups were intervened by the drug 24 h,and the flow cytometry was used to detect the apoptosis rate. The expression of Bcl-2,Bax,p-Akt,total Akt,p-PI3 K and total PI3 K were detected by Western blotting. Results: Compared with the controlled group,the inhibitory rate of cell proliferation was increased in different dose groups of Yanghe Decoction,and the inhibitory effect on cell proliferation was time and dose dependent( P < 0. 05). After 24 h of drug intervention,the apoptotic rate of MDA-MB-231 cells in each dose group was increased compared with that of the controlled group,the difference was statistically significant( P < 0. 05). Compared with the normal controlled group,the expression of Bax and Bcl-2 were decreased,and the expression of p-PI3 K was decreased relative to PI3 K,p-Aktwas decreasedrelative to Akt,and the difference was statistically significant( P < 0. 05). Conclusions: Yanghe Decoction can inhibit the proliferation and induce apoptosis of human breast cancer MDA-MB-231 cells,and its mechanism may be related to its inhibition of activation of PI3 K/Akt signaling pathway.
引文
[1]郑莹,吴春晓,张敏璐.乳腺癌在中国的流行状况和疾病特征[J].中国癌症杂志,2013,23(8):561~569
[2]窦建卫,郝云,杨亦奇,等.阳和汤治疗乳腺癌理论发微[J].世界中医药,2015,(2):196~198
[3]刘亚娴,田君.阳和汤含药血清对MCF-7细胞和MDA-MB-231细胞增殖抑制作用的实验研究[J].中华中医药学刊,2009,27(2):243~246
[4]窦建卫,任翠翠,郝云,等.阳和汤对裸鼠荷人乳腺癌组织中CD90表达的影响及其抑瘤作用[J].世界中医药,2015,(3):391~393
[5]Carey LA,Perou CM,Livasy CA,et al.Race,breast cancer subtypes,and survival in the Carolina Breast Cancer Study[J].Jama the Journal of the American Medical Association,2006,295(21):2492~502
[6]窦建卫,黄芊,赵天一,等.阳和汤对三阴性乳腺癌裸鼠移植瘤生长及IL-6、MMP-9表达的影响[J].中国现代医药杂志,2016,18(12):1~4
[7]杨连君,曹雪涛,于益芝.bcl-2,bax与肿瘤细胞凋亡[J].中国肿瘤生物治疗杂志,2003,10(3):232~234
[8]Fresno Vara J A,Casado E,De C J,et al.PI3K/Aktsignalling pathway and cancer[J].Cancer Treatment Reviews,2004,30(2):193
[2]窦建卫,郝云,杨亦奇,等.阳和汤治疗乳腺癌理论发微[J].世界中医药,2015,(2):196~198
[3]刘亚娴,田君.阳和汤含药血清对MCF-7细胞和MDA-MB-231细胞增殖抑制作用的实验研究[J].中华中医药学刊,2009,27(2):243~246
[4]窦建卫,任翠翠,郝云,等.阳和汤对裸鼠荷人乳腺癌组织中CD90表达的影响及其抑瘤作用[J].世界中医药,2015,(3):391~393
[5]Carey LA,Perou CM,Livasy CA,et al.Race,breast cancer subtypes,and survival in the Carolina Breast Cancer Study[J].Jama the Journal of the American Medical Association,2006,295(21):2492~502
[6]窦建卫,黄芊,赵天一,等.阳和汤对三阴性乳腺癌裸鼠移植瘤生长及IL-6、MMP-9表达的影响[J].中国现代医药杂志,2016,18(12):1~4
[7]杨连君,曹雪涛,于益芝.bcl-2,bax与肿瘤细胞凋亡[J].中国肿瘤生物治疗杂志,2003,10(3):232~234
[8]Fresno Vara J A,Casado E,De C J,et al.PI3K/Aktsignalling pathway and cancer[J].Cancer Treatment Reviews,2004,30(2):193