姜黄素对胰腺癌细胞增殖与凋亡的影响及其机制研究
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摘要
目的:观察姜黄素对人胰腺癌细胞株PANC-1细胞增殖及凋亡的影响,探讨姜黄素抑制胰腺癌细胞增殖及促进凋亡的分子机制。方法:常规培养人胰腺癌PANC-1细胞,采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测PANC-1细胞的增殖率,分析姜黄素浓度和处理时间对胰腺癌细胞增殖的影响;采用流式细胞仪检测PANC-1细胞凋亡率,倒置相差显微镜下观察细胞凋亡形态变化;同时检测含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)和B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)相关X蛋白(Bcl-2 associated X protein,Bax)表达变化。结果:CCK-8检测结果显示,姜黄素具有抑制胰腺癌PANC-1细胞增殖的作用,且随着姜黄素浓度的升高及处理时间的延长其抑制作用增强(P<0.05);流式细胞仪检测结果显示,姜黄素对胰腺癌PANC-1细胞具有促进凋亡作用,且随着姜黄素作用浓度的升高细胞凋亡率逐渐升高(P<0.05);实时聚合酶链反应与蛋白质印迹法结果显示,姜黄素显著增强胰腺癌PANC-1细胞Caspase-3、Caspase-9基因的表达,且呈剂量依赖性(P<0.05)。结论:姜黄素通过抑制胰腺癌细胞增殖和促进其凋亡而起到抗肿瘤效应,其潜在机制主要通过Caspases与Bcl-2通路所介导。
OBJECTIVE: To observe the effects of curcumin on the proliferation and apoptosis of human pancreatic cancer cell strain of PANC-1, and probe into the molecular mechanisms of curcumin in inhibiting the proliferation and apoptosis of PANC-1. METHODS: Human pancreatic cancer PANC-1 cells were routinely cultured, and its proliferation rate was detected by cell counting kit-8(CCK-8), so as to analyze effects of curcumin concentration and processing time on the proliferation of pancreatic cancer cells; flow cytometry was conducted to detect the apoptosis rate of PANC-1, and the morphologic change of cell apoptosis was observed under inverted phase contrast microscope; changes of expression of cysteinyl aspartate specific proteinase(Caspase) and B-cell lymphoma-2(Bcl-2) associated X protein(Bax) were detected. RESULTS: The detected results of CCK-8 indicated that curcumin can inhibiting proliferation of pancreatic cancer PANC-1, the inhibiting effect was enhanced with the increasing of curcumin concentration and the prolongation of processing time(P<0.05); results of flow cytometry showed that curcumin can promote the apoptosis of pancreatic cancer PANC-1, and the apoptosis rate was rising with the incresing of curcumin concentration(P<0.05); results of real-time polymerase chain reaction and Western Blotting indicated that curcumin can strengthen the expressions of Caspase-3 and Caspase-9 gene of PANC-1 cells, which presented a dose-dependent(P<0.05). CONCLUSIONS: Curcumin plays an anti-tumor effect by inhibiting the proliferation of pancreatic cancer cells and promoting its apoptosis, the underlying mechanism is mainly mediated by Caspases and Bcl-2 pathways.
OBJECTIVE: To observe the effects of curcumin on the proliferation and apoptosis of human pancreatic cancer cell strain of PANC-1, and probe into the molecular mechanisms of curcumin in inhibiting the proliferation and apoptosis of PANC-1. METHODS: Human pancreatic cancer PANC-1 cells were routinely cultured, and its proliferation rate was detected by cell counting kit-8(CCK-8), so as to analyze effects of curcumin concentration and processing time on the proliferation of pancreatic cancer cells; flow cytometry was conducted to detect the apoptosis rate of PANC-1, and the morphologic change of cell apoptosis was observed under inverted phase contrast microscope; changes of expression of cysteinyl aspartate specific proteinase(Caspase) and B-cell lymphoma-2(Bcl-2) associated X protein(Bax) were detected. RESULTS: The detected results of CCK-8 indicated that curcumin can inhibiting proliferation of pancreatic cancer PANC-1, the inhibiting effect was enhanced with the increasing of curcumin concentration and the prolongation of processing time(P<0.05); results of flow cytometry showed that curcumin can promote the apoptosis of pancreatic cancer PANC-1, and the apoptosis rate was rising with the incresing of curcumin concentration(P<0.05); results of real-time polymerase chain reaction and Western Blotting indicated that curcumin can strengthen the expressions of Caspase-3 and Caspase-9 gene of PANC-1 cells, which presented a dose-dependent(P<0.05). CONCLUSIONS: Curcumin plays an anti-tumor effect by inhibiting the proliferation of pancreatic cancer cells and promoting its apoptosis, the underlying mechanism is mainly mediated by Caspases and Bcl-2 pathways.
引文
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[2] 俞泽元,王科深,任彦先,等.姜黄素抗胰腺癌的作用机制.临床肝胆病杂志,2018,34(4):900-904.
[3] Jacob JN,Badyal DK,Bala S,et al.Evaluation of the in vivo anti-inflammatory and analgesic and in vitro anti-cancer activities of curcumin and its derivatives[J].Nat Prod Commun,2013,8(3):359-362.
[4] 庞慧芳,覃华,赵秋,等.姜黄素对胰腺癌细胞上皮间质转化的影响[J].世界华人消化杂志,2014,22(18):2565-2571.
[5] Guo LD,Chen XJ,Hu YH,et al.Curcumin inhibits proliferation and induces apoptosis of human colorectal cancer cells by activating the mitochondria apoptotic pathway[J].Phytother Res,2013,27(3):422-430.
[6] 赵轶峰,魏玉磊,王萍,等.姜黄素抑制 Wnt /β-catenin信号通路诱导乳腺癌细胞凋亡的机制研究[J].现代中西医结合杂志,2016,25(29):3202-3204,3306.
[7] 金宇斌,李立德,韩莹,等.姜黄素调控食管癌Eca-109细胞分化、增殖、凋亡的机制[J].中国老年学杂志,2017,37(4):806-809.
[8] 王巧琳,何娜娜,陆明,等.姜黄素对乳腺癌细胞的增殖与迁移及侵袭的影响及其作用机制[J].中国临床药理学杂志,2017,33(18):1793-1796.
[9] 陈永宏,高月.姜黄素对非小细胞肺癌A549细胞增殖凋亡和侵袭能力的影响[J].中国药业,2018,27(23):9-12.
[10] 李伟锋,蒋建兰.姜黄素药理作用的研究现状[J].中国临床药理学杂志,2017,33(10):957-960.
[11] 周晶晶,郑昱辰,李明月,等.姜黄素的药理作用研究进展[J].吉林医药学院学报,2016,37(4):304-307.
[12] Jutooru I,Chadalapaka G,Lei P,et al.Inhibition of NF kappa B and pancreatic cancer cell and tumor growth by curcumin is dependent on specificity protein down-regulation[J].J Biol Chem,2010,285(33):25332-25344.
[13] Sahu RP,Batra S,Srivastava SK.Activation of ATM/Chk1 by curcumin causes cell cycle arrest and apoptosis in human pancreatic cancer cells[J].Br J Cancer,2009,100(9):1425-1433.
[14] 罗长江,俞泽元,曹宏泰,等.姜黄素与吉西他滨联用对胰腺癌细胞株增殖及凋亡的诱导作用[J].中华实验外科杂志,2018,35(8):1413-1415.