斑蝥酸钠抑制人结肠癌细胞增殖及与STAT3信号通路的关系
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摘要
目的探讨斑蝥酸钠对人结肠癌细胞增殖、凋亡的影响及其与信号转导子与转录激活子3(STAT3)信号通路的关系。方法人结肠癌SW480、HCT116细胞分为实验组和对照组,实验组细胞分别以0. 5,1. 0,2. 0,4. 0μg·m L~(-1)斑蝥酸钠维生素B6注射液处理,对照组则以等量生理盐水处理。24 h后于光学倒置显微镜下观察细胞形态,以噻唑蓝(MTT)法检测细胞增殖能力,以流式细胞术检测细胞凋亡率,以蛋白免疫印迹(WB)法检测细胞STAT3、磷酸化-STAT3(p-STAT3)、B淋巴细胞瘤-2基因(bcl-2)、C-myc蛋白表达情况。结果干预48 h后,0. 5,1. 0,2. 0,4. 0μg·m L~(-1)实验组SW480细胞增殖抑制率分别为(23. 05±0. 14)%,(33. 71±0. 15)%,(49. 89±0. 09)%,(73. 05±0. 11)%;HCT116细胞增殖抑制率分别为(21. 54±0. 12)%,(31. 55±0. 09)%,(43. 24±0. 15)%,(71. 76±0. 18)%。对照组及低、中、高剂量实验组(1. 0,2. 0,4. 0μg·m L~(-1))人结肠癌SW480细胞凋亡率分别为(1. 46±0. 22)%,(17. 49±1. 21)%,(40. 01±2. 19)%,(68. 83±3. 82)%,HCT116细胞凋亡率分别为(1. 01±0. 13)%,(14. 13±1. 25)%,(36. 35±1. 67)%,(62. 16±3. 31)%,组间差异均有统计学意义(均P <0. 05)。WB检测显示,与对照组比较,低、中、高剂量实验组SW480、HCT116细胞STAT3、p-STAT3、bcl-2、C-myc蛋白相对表达量显著降低(均P <0. 05)。结论斑蝥酸钠可抑制人结肠癌细胞增殖,诱导细胞凋亡,并呈现一定的剂量依赖性,与其可降低STAT3信号通路蛋白表达有关。
Objective To investigate the effect of sodium cantharidate on proliferation and apoptosis of human colon cancer cells and its relationship with signal transducer and activator of transcription 3( STAT3)signaling pathway. Methods Human colon cancer SW480 and HCT116 cells were divided into experimental groups and control group. Cells in experimental groups were treated with 0. 5,1. 0,2. 0 and 4. 0 μg·m L~(-1) sodium cantharidate and vitamin B6 injection respectively,while those in the control group were treated with the same amount of normal saline. After 48 h,cell morphology was observed under optical inversion microscope. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect apoptotic rate. Western blot( WB) were used to detect the expression of STAT3,phosphorylated STAT3( p-STAT3),B lymphoma-2 gene( bcl-2) and C-myc protein. Results After 48 h of drug intervention,the inhibition rates of proliferation of SW480 cells in the experimental groups( 0. 5,1. 0,2. 0,4. 0 μg·mL~(-1)) were( 23. 05 ± 0. 14) %,( 33. 71 ± 0. 15) %,( 49. 89 ± 0. 09) %,( 73. 05 ± 0. 11) %, and which were( 21. 54 ± 0. 12) %,( 31. 55 ± 0. 09) %,( 43. 24 ± 0. 15) %,( 71. 76 ± 0. 18) % in the HCT116 cells respectively; the apoptotic rates of human colon cancer SW480 cells in control group and low,medium and high dose experimental groups( 1. 0,2. 0,4. 0μg·mL~(-1)) were( 1. 46 ± 0. 22) %,( 17. 49 ± 1. 21) %,( 40. 01 ± 2. 19) %,( 68. 83 ± 3. 82) %,which were( 1. 01 ± 0. 13) %,( 14. 13 ± 1. 25) %,( 36. 35 ± 1. 67) %,( 62. 16 ± 3. 31) % in the HCT116 cells,there was significant difference( all P < 0. 05). WB assay showed that the relative expression of STAT3,p-STAT3,Bcl-2 and C-myc protein in SW480 and HCT116 cells in low,medium and high dose experimental group were significantly lower than those in control group( all P < 0. 05). Conclusion Sodium cantharidin can inhibit the proliferation of human colon cancer cells and induce apoptosis in a dose-dependent manner,which is related to the reduction of STAT3 signaling pathway protein expression.
Objective To investigate the effect of sodium cantharidate on proliferation and apoptosis of human colon cancer cells and its relationship with signal transducer and activator of transcription 3( STAT3)signaling pathway. Methods Human colon cancer SW480 and HCT116 cells were divided into experimental groups and control group. Cells in experimental groups were treated with 0. 5,1. 0,2. 0 and 4. 0 μg·m L~(-1) sodium cantharidate and vitamin B6 injection respectively,while those in the control group were treated with the same amount of normal saline. After 48 h,cell morphology was observed under optical inversion microscope. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect apoptotic rate. Western blot( WB) were used to detect the expression of STAT3,phosphorylated STAT3( p-STAT3),B lymphoma-2 gene( bcl-2) and C-myc protein. Results After 48 h of drug intervention,the inhibition rates of proliferation of SW480 cells in the experimental groups( 0. 5,1. 0,2. 0,4. 0 μg·mL~(-1)) were( 23. 05 ± 0. 14) %,( 33. 71 ± 0. 15) %,( 49. 89 ± 0. 09) %,( 73. 05 ± 0. 11) %, and which were( 21. 54 ± 0. 12) %,( 31. 55 ± 0. 09) %,( 43. 24 ± 0. 15) %,( 71. 76 ± 0. 18) % in the HCT116 cells respectively; the apoptotic rates of human colon cancer SW480 cells in control group and low,medium and high dose experimental groups( 1. 0,2. 0,4. 0μg·mL~(-1)) were( 1. 46 ± 0. 22) %,( 17. 49 ± 1. 21) %,( 40. 01 ± 2. 19) %,( 68. 83 ± 3. 82) %,which were( 1. 01 ± 0. 13) %,( 14. 13 ± 1. 25) %,( 36. 35 ± 1. 67) %,( 62. 16 ± 3. 31) % in the HCT116 cells,there was significant difference( all P < 0. 05). WB assay showed that the relative expression of STAT3,p-STAT3,Bcl-2 and C-myc protein in SW480 and HCT116 cells in low,medium and high dose experimental group were significantly lower than those in control group( all P < 0. 05). Conclusion Sodium cantharidin can inhibit the proliferation of human colon cancer cells and induce apoptosis in a dose-dependent manner,which is related to the reduction of STAT3 signaling pathway protein expression.
引文
[1]卢志坤,吴永强,何德谋,等.直肠癌Miles手术行经腹直肌乙状结肠造口对减少术后结肠造口旁疝发生率的作用[J].中华疝和腹壁外科杂志(电子版),2016,10(2):125-127.
[2]孟慧,孙旭,念家云,等.斑蝥酸钠联合化疗治疗进展期胃癌的Meta分析[J].世界中医药,2017,12(10):2496-2500.
[3]黄秋兰,雷荣娥,覃沁怡,等.白细胞介素9通过激活信号转导子和转录激活子3(STAT3)通路促进胰腺癌细胞的增殖和迁移[J].细胞与分子免疫学杂志,2017,21(9):1228-1233.
[4]郭伟健,何敏毅,孙学刚,等.雷公藤甲素对大肠癌细胞细胞周期的调控作用[J].贵州医科大学学报,2012,37(2):141-144.
[5]张丽,朱蕾.葛根素抑制人子宫颈癌细胞株He La细胞增殖研究[J].贵州医药,2017,41(3):242-244.
[6]唐瑶,包晓霞,鲁周南,等.“连花汤”正丁醇提物对子宫内膜癌细胞增殖和凋亡的影响[J].环球中医药,2017,10(6):592-596.
[7]张添昊,任艳玲,艾思羽,等.左归丸和右归丸对去卵巢大鼠股骨骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达的影响[J].中医杂志,2017,58(6):511-515.
[8]施庆彤,刁亚利,包阳.斑蝥酸钠维生素B6注射液对非小细胞肺癌手术患者血清VEGF水平的影响[J].中国现代医学杂志,2017,27(7):61-63.
[9] FRIAS,MIGUEL A,JAMES,et al. Native and reconstituted HDL activate Stat3 in ventricular cardiomyocytes via ERK1/2:role of sphingosine-1-phosphate[J]. Cardiol Res,2017,82(2):313-321.
[10] AXELROD J H,LANTON T,SHRIKI A,et al. P0245:The IL-6/SIL6R and STAT3 axis is crucial for hepatoprotection and suppression of liver cancer in female mice with chronic liver injury[J]. J Hepatol,2015,62(23):398.
[2]孟慧,孙旭,念家云,等.斑蝥酸钠联合化疗治疗进展期胃癌的Meta分析[J].世界中医药,2017,12(10):2496-2500.
[3]黄秋兰,雷荣娥,覃沁怡,等.白细胞介素9通过激活信号转导子和转录激活子3(STAT3)通路促进胰腺癌细胞的增殖和迁移[J].细胞与分子免疫学杂志,2017,21(9):1228-1233.
[4]郭伟健,何敏毅,孙学刚,等.雷公藤甲素对大肠癌细胞细胞周期的调控作用[J].贵州医科大学学报,2012,37(2):141-144.
[5]张丽,朱蕾.葛根素抑制人子宫颈癌细胞株He La细胞增殖研究[J].贵州医药,2017,41(3):242-244.
[6]唐瑶,包晓霞,鲁周南,等.“连花汤”正丁醇提物对子宫内膜癌细胞增殖和凋亡的影响[J].环球中医药,2017,10(6):592-596.
[7]张添昊,任艳玲,艾思羽,等.左归丸和右归丸对去卵巢大鼠股骨骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达的影响[J].中医杂志,2017,58(6):511-515.
[8]施庆彤,刁亚利,包阳.斑蝥酸钠维生素B6注射液对非小细胞肺癌手术患者血清VEGF水平的影响[J].中国现代医学杂志,2017,27(7):61-63.
[9] FRIAS,MIGUEL A,JAMES,et al. Native and reconstituted HDL activate Stat3 in ventricular cardiomyocytes via ERK1/2:role of sphingosine-1-phosphate[J]. Cardiol Res,2017,82(2):313-321.
[10] AXELROD J H,LANTON T,SHRIKI A,et al. P0245:The IL-6/SIL6R and STAT3 axis is crucial for hepatoprotection and suppression of liver cancer in female mice with chronic liver injury[J]. J Hepatol,2015,62(23):398.