胡桃醌通过线粒体途径抑制宫颈癌Hela细胞生长的研究
|
摘要
胡桃醌的化学成分为5-羟基-1,4-萘二酮,是从核桃楸中分离出的重要化合物,对多种恶性肿瘤的生长具有抑制活性。研究中,子宫颈癌Hela细胞接受不同剂量的胡桃醌处理,利用MTT实验检测胡桃醌对Hela细胞的增殖抑制作用、流式细胞术检测胡桃醌对Hela细胞周期和凋亡的影响以及Western blot检测CytC、Bcl-2、Bax和caspases蛋白水平的表达。结果表明,胡桃醌可以呈剂量依赖性的方式显著抑制Hela细胞的活性。此外,膜联蛋白Annexin V-FITC在50μM和100μM浓度的早期凋亡率分别为16. 79%和30. 03%,而对照组为10. 73%。不同剂量胡桃醌处理Hela细胞后,Cytc水平和Bax/Bcl-2比值较对照组明显增高,同时caspase-3、-9的活性也明显变化。结果表明,胡桃醌可能通过线粒体途径诱导细胞凋亡有效抑制Hela细胞的生长。
The chemical component of juglone is 5-hydroxy-1,4-naphthalenedione,which is major chemical constituent of Juglans mandshurica Maxim and confirmed to possesses anti-tumor activity on the growth of a variety of malignant tumors. In this study,cervical cancer Hela were cultured and treated with different dosages of juglone. MTT assay was performed to examine the proliferation inhibition of juglone on Hela cells. Flow cytometry was used to study juglone on the cell cycle and apoptosis of juglone-treated Hela cells. The protein expression levels of CytC,Bcl-2,Bax and caspases were measured by Western blot. The results showed that juglone significantly inhibited Hela cells survival in a dose-dependent manner. Moreover,the percentages of early apoptosis of Annexin V-FITC were 16. 79% and 30. 03% in 50 μM and 100 μM groups,compared with 10. 73% in the control group,respectively. After cells were treated at different doses,juglone increased Cytc level and Bax/Bcl-2 ratio significantly compared with those of the control group. These events paralleled with activation of caspase-3 and caspase-9. The results suggest that juglone may be effective for the treatment of Hela cells and probably induce apoptosis through mitochondrial pathway.
The chemical component of juglone is 5-hydroxy-1,4-naphthalenedione,which is major chemical constituent of Juglans mandshurica Maxim and confirmed to possesses anti-tumor activity on the growth of a variety of malignant tumors. In this study,cervical cancer Hela were cultured and treated with different dosages of juglone. MTT assay was performed to examine the proliferation inhibition of juglone on Hela cells. Flow cytometry was used to study juglone on the cell cycle and apoptosis of juglone-treated Hela cells. The protein expression levels of CytC,Bcl-2,Bax and caspases were measured by Western blot. The results showed that juglone significantly inhibited Hela cells survival in a dose-dependent manner. Moreover,the percentages of early apoptosis of Annexin V-FITC were 16. 79% and 30. 03% in 50 μM and 100 μM groups,compared with 10. 73% in the control group,respectively. After cells were treated at different doses,juglone increased Cytc level and Bax/Bcl-2 ratio significantly compared with those of the control group. These events paralleled with activation of caspase-3 and caspase-9. The results suggest that juglone may be effective for the treatment of Hela cells and probably induce apoptosis through mitochondrial pathway.
引文
[1]SAEI GHARE NAZ M,KARIMAN N,EBADI A,et al.Educational interventions for cervical cancer screening behavior of women:A systematic review[J].Asian Pac JCancer Prev.2018,19(4):875-884.
[2]GONG J M,SHEN Y,SHAN W W,et al.The association between MTHFR polymorphism and cervical cancer[J].Sci Rep,2018,8(1):7244.
[3]MA J Y,KE L C,LIU Q.The pretreatment platelet-to-lymphocyte ratio predicts clinical outcomes in patients with cervical cancer:A meta-analysis[J].Medicine(Baltimore),2018,97(43):e12897.
[4]LU Z,CHEN H,ZHENG X M,et al.Experimental study on the apoptosis of cervical cancer Hela cells induced by juglone through c-Jun N-terminal kinase/cJun pathway[J].Asian Pac J Trop Med,2017,10(6):572-575.
[5]WANG J,LIU K,WANG X F,et al.Juglone reduces growth and migration of U251 glioblastoma cells and disrupts angiogenesis[J].Oncol Rep,2017,38(4):1959-1966.
[6]ZHANG X B,ZOU C L,DUAN Y X,et al.Activity guided isolation and modification of juglone from Juglans regia as potent cytotoxicagent against lung cancer cell lines[J].BMC Complement Altern Med,2015,15:396.
[7]FANG F,QIN Y,QI L,et al.Juglone exerts antitumor effect in ovarian cancer cells[J].Iran J Basic Med Sci,2015,18(6):544-548.
[8]LIN L,PIAO J,MA Y,et al.Mechanisms underlying cancer growth and apoptosis by DEK overexpression in colorectal cancer[J].PLo S One,2014,9(10):e111260.
[9]OURIQUE F,KVIECINSKI M R,ZIRBEL G,et al.In vivo inhibition of tumor progression by 5 hydroxy-1,4-naphthoquinone(juglone)and 2-(4-hydroxyanilino)-1,4-naphthoquinone(Q7)in combination with ascorbate[J].Biochem Biophys Res Commun,2016,477(4):640-646.
[10]WU J,ZHANG H,XU Y,et al.Juglone induces apoptosis of tumor stem-like cells through ROS-p38 pathway in glioblastoma[J].BMC Neurol,2017,17(1):70.
[11]JIN X,ZHANG Y,ZHANG Z,et al.Juglone loaded poloxamer 188/phospholipid mixed micelles evaluated in vitro and in vivo in breast cancer[J].Int J Pharm,2016,515(1-2):359-366.
[12]ABYAR F,TABIZI L.Experimental and theoretical investigations of novel oxidovanadium(IV)juglone complex:DNA/HSA interaction and cytotoxic activity[J].JBiomol Struct Dyn,2019,4:1-14.
[13]KIM J A,LEE E K,PARK S J,et al.Novel anticancer role of naphthazarin in human gastric cancer cells[J].Int J Oncol,2012,40(1):157-162.
[14]MANICKAM P,KAUSHIK A,KARUNAKARAN C,et al.Recent advances in cytochrome c biosensing technologies[J].Biosens Bioelectron,2017,87:654-668.
[15]RADOGNA F,CRISTOFANON S,PATENOSTER L,et al.Melatonin antagonizes the intrinsic pathway of apoptosis via mitochondrial targeting of Bcl-2[J].J Pineal Res,2008,44(3):316-325.
[16]GUPTA R,GHOSH S.Bax and Bif-1 proteins interact on bilayer lipid membrane and form pore[J].Biochem Biophys Res Commun,2015,463(4):751-755.
[17]LOVSCHALL H,KASSEM M,MOSEKILDE L.Apoptosis:molecular aspect[J].Nord Med,1997,112(8):271-275.
[18]PASTUSZKO P,SCHEARS G J,KUBIN J,et al.Granulocyte colony-stimulating factor significantly decreases density of hippocampal caspase 3-positive nuclei,thus ameliorating apoptosis-mediated damage,in a model of ischaemic neonatal brain injury[J].Interact Cardiovasc Thorac Surg,2017,25(4):600-605.
[19]KIM J S,HA J Y,YANG S J,et al.A novel non-apoptotic role of procaspase-3 in the regulation of mitochondrial biogenesis activators[J].J Cell Biochem,2018,119(1):347-357.
[20]ZHANG S S,LIAO Z X,HUANG R Z,et al.A new aromatic glycoside and its anti-proliferative activities from the leaves of Bergenia purpurascens[J].Nat Prod Res,2018,32(6):668-675.
[2]GONG J M,SHEN Y,SHAN W W,et al.The association between MTHFR polymorphism and cervical cancer[J].Sci Rep,2018,8(1):7244.
[3]MA J Y,KE L C,LIU Q.The pretreatment platelet-to-lymphocyte ratio predicts clinical outcomes in patients with cervical cancer:A meta-analysis[J].Medicine(Baltimore),2018,97(43):e12897.
[4]LU Z,CHEN H,ZHENG X M,et al.Experimental study on the apoptosis of cervical cancer Hela cells induced by juglone through c-Jun N-terminal kinase/cJun pathway[J].Asian Pac J Trop Med,2017,10(6):572-575.
[5]WANG J,LIU K,WANG X F,et al.Juglone reduces growth and migration of U251 glioblastoma cells and disrupts angiogenesis[J].Oncol Rep,2017,38(4):1959-1966.
[6]ZHANG X B,ZOU C L,DUAN Y X,et al.Activity guided isolation and modification of juglone from Juglans regia as potent cytotoxicagent against lung cancer cell lines[J].BMC Complement Altern Med,2015,15:396.
[7]FANG F,QIN Y,QI L,et al.Juglone exerts antitumor effect in ovarian cancer cells[J].Iran J Basic Med Sci,2015,18(6):544-548.
[8]LIN L,PIAO J,MA Y,et al.Mechanisms underlying cancer growth and apoptosis by DEK overexpression in colorectal cancer[J].PLo S One,2014,9(10):e111260.
[9]OURIQUE F,KVIECINSKI M R,ZIRBEL G,et al.In vivo inhibition of tumor progression by 5 hydroxy-1,4-naphthoquinone(juglone)and 2-(4-hydroxyanilino)-1,4-naphthoquinone(Q7)in combination with ascorbate[J].Biochem Biophys Res Commun,2016,477(4):640-646.
[10]WU J,ZHANG H,XU Y,et al.Juglone induces apoptosis of tumor stem-like cells through ROS-p38 pathway in glioblastoma[J].BMC Neurol,2017,17(1):70.
[11]JIN X,ZHANG Y,ZHANG Z,et al.Juglone loaded poloxamer 188/phospholipid mixed micelles evaluated in vitro and in vivo in breast cancer[J].Int J Pharm,2016,515(1-2):359-366.
[12]ABYAR F,TABIZI L.Experimental and theoretical investigations of novel oxidovanadium(IV)juglone complex:DNA/HSA interaction and cytotoxic activity[J].JBiomol Struct Dyn,2019,4:1-14.
[13]KIM J A,LEE E K,PARK S J,et al.Novel anticancer role of naphthazarin in human gastric cancer cells[J].Int J Oncol,2012,40(1):157-162.
[14]MANICKAM P,KAUSHIK A,KARUNAKARAN C,et al.Recent advances in cytochrome c biosensing technologies[J].Biosens Bioelectron,2017,87:654-668.
[15]RADOGNA F,CRISTOFANON S,PATENOSTER L,et al.Melatonin antagonizes the intrinsic pathway of apoptosis via mitochondrial targeting of Bcl-2[J].J Pineal Res,2008,44(3):316-325.
[16]GUPTA R,GHOSH S.Bax and Bif-1 proteins interact on bilayer lipid membrane and form pore[J].Biochem Biophys Res Commun,2015,463(4):751-755.
[17]LOVSCHALL H,KASSEM M,MOSEKILDE L.Apoptosis:molecular aspect[J].Nord Med,1997,112(8):271-275.
[18]PASTUSZKO P,SCHEARS G J,KUBIN J,et al.Granulocyte colony-stimulating factor significantly decreases density of hippocampal caspase 3-positive nuclei,thus ameliorating apoptosis-mediated damage,in a model of ischaemic neonatal brain injury[J].Interact Cardiovasc Thorac Surg,2017,25(4):600-605.
[19]KIM J S,HA J Y,YANG S J,et al.A novel non-apoptotic role of procaspase-3 in the regulation of mitochondrial biogenesis activators[J].J Cell Biochem,2018,119(1):347-357.
[20]ZHANG S S,LIAO Z X,HUANG R Z,et al.A new aromatic glycoside and its anti-proliferative activities from the leaves of Bergenia purpurascens[J].Nat Prod Res,2018,32(6):668-675.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |