从STAT3通路探讨桂枝茯苓丸对MCF-7细胞增殖的抑制作用机制
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摘要
目的探讨桂枝茯苓丸抑制乳腺癌MCF-7细胞增殖作用机制。方法采用MTT法检测高低剂量桂枝茯苓丸(1800g/m L、2700g/m L)及其与STAT3通路抑制剂AG490联合作用48h后对MCF-7细胞增殖抑制作用;流式细胞术检测细胞周期及细胞中STAT3、P-STAT3、Cyclin D1、P53表达的变化。结果与空白组比较,用药组抑制细胞增殖作用呈剂量依赖性,加阻断剂AG490联合用药组明显高于相对应的单独用药组。与空白对照组比较,AG490组G0/G1期细胞比例升高明显(P <0. 05),桂枝茯苓丸高、低剂量组及其联合用药组G0/G1期和S期细胞比例降低(P <0. 05)。与空白组比较,各组P-STAT3的表达都显著下调(P <0. 05),但各剂量组STAT3表达均无显著变化(P> 0. 05);除低剂量桂枝茯苓丸组无差异外,其它各组P53表达显著上调(P <0. 05);只有高剂量桂枝茯苓丸组及其与AG490联合组Cyclin D1表达显著下调(P <0. 05)。与相应剂量桂枝茯苓丸相比,加AG490阻断剂后,P-STAT3、Cyclin D1表达均下调明显,P53表达均上调明显。结论桂枝茯苓丸及STAT3通路AG490阻断剂能抑制MCF-7细胞生长,且呈剂量依赖性,其机理与药物调控P-STAT3、Cyclin D1的表达下调及蛋白P53表达上调有关。
Objective To study the effect of Guizhi Fuling pill on the growth of MCF-7 cells from human breast cancer and the protein expression of STAT3,p-STAT3,cyclin D1,and p53,and to explore the mechanism of Guizhi Fuling pill on inhibiting the proliferation of MCF-7 breast cancer cells,and which provide a theoretical foundation in the clinical application.Methods The inhibitory effect of the high and low dose of Guizhi Fuling pill( 1800 g/m L、2700 g/m L) and its combination with the inhibitor AG490 of STAT3 pathway after 48 h on the proliferation of MCF-7 cell was detected by MTT. The cell of cycle and the changes of STAT3,p-STAT3,cyclin D1 and p53 expression in cells were study by flow cytometry. Results Compared with the blank group,the inhibitory effect of the drug group on cell was dose-dependent,and the group of inhibitor AG490 was significantly higher than the corresponding drug group. Compared with the blank group,the cells were blocked in G0/G1 phase mixed with AG490( P < 0. 05); and the ratio of G0/G1 and S phase could be decreased in the high,low concentration group of Guizhi Fuling pill and the its combined AG490 group( P < 0. 05). Compared with the blank group,the expression of p-STAT3 was significantly reduced( P < 0. 05),but no significant changes in the expression of STAT3 in each dose group; Except for the low dose of Guizhi Fuling pill group,the expression of p53 was significantly increased in other groups( P < 0. 05); Only the expression of cyclin D1 in the high dose of Guizhi Fuling pill group and its combined group with AG490 were significantly reduced( P <0. 05); After combining AG490,the expression of p-STAT3 and cyclin D1 were reduced significantly,and the expression of p53 was significantly increased compared with the same dose group of Guizhi Fuling pill( P < 0. 05). Conclusion Guizhi Fuling pill and the inhibitor AG490 of STAT3 pathway can inhibit the growth of MCF-7 cells,and is dose – dependent,which mechanism is related to the down-regulation of the expression of p-STAT3,cyclin D1 and the up-regulation of protein p53 expression.
Objective To study the effect of Guizhi Fuling pill on the growth of MCF-7 cells from human breast cancer and the protein expression of STAT3,p-STAT3,cyclin D1,and p53,and to explore the mechanism of Guizhi Fuling pill on inhibiting the proliferation of MCF-7 breast cancer cells,and which provide a theoretical foundation in the clinical application.Methods The inhibitory effect of the high and low dose of Guizhi Fuling pill( 1800 g/m L、2700 g/m L) and its combination with the inhibitor AG490 of STAT3 pathway after 48 h on the proliferation of MCF-7 cell was detected by MTT. The cell of cycle and the changes of STAT3,p-STAT3,cyclin D1 and p53 expression in cells were study by flow cytometry. Results Compared with the blank group,the inhibitory effect of the drug group on cell was dose-dependent,and the group of inhibitor AG490 was significantly higher than the corresponding drug group. Compared with the blank group,the cells were blocked in G0/G1 phase mixed with AG490( P < 0. 05); and the ratio of G0/G1 and S phase could be decreased in the high,low concentration group of Guizhi Fuling pill and the its combined AG490 group( P < 0. 05). Compared with the blank group,the expression of p-STAT3 was significantly reduced( P < 0. 05),but no significant changes in the expression of STAT3 in each dose group; Except for the low dose of Guizhi Fuling pill group,the expression of p53 was significantly increased in other groups( P < 0. 05); Only the expression of cyclin D1 in the high dose of Guizhi Fuling pill group and its combined group with AG490 were significantly reduced( P <0. 05); After combining AG490,the expression of p-STAT3 and cyclin D1 were reduced significantly,and the expression of p53 was significantly increased compared with the same dose group of Guizhi Fuling pill( P < 0. 05). Conclusion Guizhi Fuling pill and the inhibitor AG490 of STAT3 pathway can inhibit the growth of MCF-7 cells,and is dose – dependent,which mechanism is related to the down-regulation of the expression of p-STAT3,cyclin D1 and the up-regulation of protein p53 expression.
引文
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[13] Jin MS,Park IA. New insight on the biological role of p53 protein as atumor suppressor:re-evaluation of its clinical significancein triple-negative breast cancer[J]. Tumor Biology,2016,37(8):11017.
[14]马丽萍,陈德喜.转染野生型p53对乳腺癌细胞所致细胞凋亡的研究[J].肿瘤,2000,20(2):100.
[15]李桂青. HSP60和p53在胃癌中的表达及意义[J].临床消化病杂志,2014,26(1):11.
[16]杨会杰,宋金凤,魏民. IMP3、p53在宫颈癌中的表达及临床意义[J].现代妇产科进展,2013,22(4):320..
[17] Sheng HY,Yao D,Zhu CH. Transglutaminase2 Inhibitor KCC009 In-duces p53-Independent Radiosensitization in lung Adenocarcinomacells[J]. Medical Science Monitor,2016,22:5041.
[2]郭晓娟,韩立.桂枝茯苓丸联用顺铂紫杉醇化疗提高卵巢癌多药耐药模型裸鼠生存率[J].科学技术与工程,2016,16(20):120.
[3]古妻.国外医学.中医中药分册[M].北京:中国中医研究院图书情报研究所,1987:159.
[4]李澜.桂枝茯苓丸抑制人乳腺癌细胞MCF-7增殖诱导细胞周期阻滞的作用机制研究[D].河南中医学院硕士学位论文,2015.
[5]夏雷达,蒋时红.桂枝茯苓丸治疗妇科肿瘤的研究概况[J].中医研究,2016,29(5):78.
[6]刘松江,韩淑丽.桂枝茯苓丸异病同治的临床应用[J].中医药信息,2014,31(3):34.
[7]余微波,谷俊朝.乳腺癌与其他激素关系的研究进展[J].国外医学·外科学2005,32(2):122.
[8]张文,王雅杰. STAT3与乳腺癌的研究进展[J].现代肿瘤医学,2007,15(3):439.
[9]张永华,孙治君.表皮生长因子受体与乳腺癌的研究进展[J].现代肿瘤医学,2011,19(7):1454.
[10]王前,邓晶,蒋永新,等. CyclinD1的研究进展[J].现代肿瘤医学,2009,17(2):350.
[11] Williams,GH,Stoeber,Kai. The cell cycle and cancer[J]. Journal ofPathology,2012,226(2):352.
[12]李会平. p27、PCNA、Cyclin D1等因子的表达和乳腺癌早期浸润的关系[D].河南科技大学硕士学位论文,2010.
[13] Jin MS,Park IA. New insight on the biological role of p53 protein as atumor suppressor:re-evaluation of its clinical significancein triple-negative breast cancer[J]. Tumor Biology,2016,37(8):11017.
[14]马丽萍,陈德喜.转染野生型p53对乳腺癌细胞所致细胞凋亡的研究[J].肿瘤,2000,20(2):100.
[15]李桂青. HSP60和p53在胃癌中的表达及意义[J].临床消化病杂志,2014,26(1):11.
[16]杨会杰,宋金凤,魏民. IMP3、p53在宫颈癌中的表达及临床意义[J].现代妇产科进展,2013,22(4):320..
[17] Sheng HY,Yao D,Zhu CH. Transglutaminase2 Inhibitor KCC009 In-duces p53-Independent Radiosensitization in lung Adenocarcinomacells[J]. Medical Science Monitor,2016,22:5041.