白杨素对口腔鳞状细胞癌KB细胞凋亡的影响及其机制
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摘要
目的:探讨白杨素对口腔鳞状细胞癌KB细胞凋亡的影响及其机制,为临床上口腔鳞状细胞癌的治疗提供思路。方法:用不同浓度白杨素(1,2,4,8,16和32μmol/L)处理KB细胞24 h,采用MMT法检测细胞增殖,流式细胞术检测细胞凋亡,化学发光法检测caspase-3/7活性,JC-1法检测KB细胞线粒体膜电位的变化,蛋白质印迹检测蛋白激酶B(protein kinase B,AKT)和磷脂酰肌醇3-激酶(phosphoinositide-3-kinase,PI3K)的活化。结果:白杨素以浓度依赖方式抑制KB细胞增殖并诱导其凋亡,促进caspase-3/7的活化,降低KB细胞线粒体膜电位;同时抑制AKT和PI3K磷酸化。结论:白杨素诱导KB细胞凋亡作用可能与线粒体功能障碍和抑制PI3K/AKT通路相关。
Objective: To investigate the effect of chrysin on apoptosis of oral squamous carcinoma KB cell line and the possible mechanisms, and to provide new ideas for the treatment of oral cancer.Methods: Oral cancer KB cells were treated with different concentrations of chrysin(1, 2, 4, 8, 16, and 32 μmol/L) for 24 h. Cell proliferation was detected by MMT assay; apoptosis was detected by flow cytometry; the activity of caspase-3/7 was detected by chemiluminescent assay; mitochondrial membrane potential in KB cells was determined by JC-1 assay; and Western blotting was used to determine the activation of protein kinase B(AKT) and phosphoinositide-3-kinase(PI3 K).Results: Chrysin inhibited the proliferation of KB cells in a concentration-dependent manner,accompanied by increase in apoptosis of KB cells, activation of caspase-3/7, decrease in mitochondrial membrane potential, and suppression of the phosphorylation of AKT and PI3 K.Conclusion: The effect of chrysin on KB cell apoptosis may be related to mitochondrial dysfunction and inhibition of PI3 K/AKT pathway.
Objective: To investigate the effect of chrysin on apoptosis of oral squamous carcinoma KB cell line and the possible mechanisms, and to provide new ideas for the treatment of oral cancer.Methods: Oral cancer KB cells were treated with different concentrations of chrysin(1, 2, 4, 8, 16, and 32 μmol/L) for 24 h. Cell proliferation was detected by MMT assay; apoptosis was detected by flow cytometry; the activity of caspase-3/7 was detected by chemiluminescent assay; mitochondrial membrane potential in KB cells was determined by JC-1 assay; and Western blotting was used to determine the activation of protein kinase B(AKT) and phosphoinositide-3-kinase(PI3 K).Results: Chrysin inhibited the proliferation of KB cells in a concentration-dependent manner,accompanied by increase in apoptosis of KB cells, activation of caspase-3/7, decrease in mitochondrial membrane potential, and suppression of the phosphorylation of AKT and PI3 K.Conclusion: The effect of chrysin on KB cell apoptosis may be related to mitochondrial dysfunction and inhibition of PI3 K/AKT pathway.
引文
[1]Chen YJ,Lin SC,Kao T,et al.Genome-wide profiling of ora squamous cell carcinoma[J].J Pathol,2004,204(3):326-332.
[2]Lo WL,Kao SY,Chi LY,et al.Outcomes of oral squamous cel carcinoma in Taiwan after surgical therapy:factors affecting survival[J].J Oral Maxillofac Surg,2003,61(7):751-758.
[3]Kasala ER,Bodduluru LN,Madana RM,et al.Chemopreventive and therapeutic potential of chr ysin in cancer:mechanistic perspectives[J].Toxicol Lett,2015,233(2):214-225.
[4]Nile SH,Keum YS,Nile AS,et al.Antioxidant,anti-inflammatory and enzyme inhibitory activity of natural plant flavonoids and their synthesized derivatives[J].J Biochem Mol Toxicol,2018,32(1)e22002.
[5]Khoo BY,Chua SL,Balaram P.Apoptotic effects of chrysin in human cancer cell lines[J].Int J Mol Sci,2010,11(5):2188-2199.
[6]Wang J,Qiu J,Dong J,et al.Chr ysin protects mice from Staphylococcus aureus pneumonia[J].J Appl Microbiol,2011111(6):1551-1558.
[7]Du Q,Gu X,Cai J,et al.Chr ysin attenuates allergic air way inflammation by modulating the transcription factors T-bet and GATA-3 in mice[J].Mol Med Rep,2012,6(1):100-104.
[8]Chen YH,Yang ZS,Wen CC,et al.Evaluation of the structureactivity relationship of flavonoids as antioxidants and toxicants o zebrafish larvae[J].Food Chem,2012,134(2):717-724.
[9]Gresa-arribas N,Serratosa J,Saura J,et al.Inhibition of CCAAT/enhancer binding protein delta expression by chrysin in microglia cells results in anti-inflammatory and neuroprotective effects[J].JNeurochem,2010,115(2):526-536.
[10]Lopez J,Tait SW.Mitochondrial apoptosis:killing cancer using the enemy within[J].Br J Cancer,2015,112(6):957-962.
[11]Yoon TM,Kim SA,Lee DH,et al.EGR1 regulates radiation-induced apoptosis in head and neck squamous cell carcinoma[J].Oncol Rep2015,33(4):1717-1722.
[12]Park W,Park S,Lim W,et al.Chr ysin disrupts intracellular homeostasis through mitochondria-mediated cell death in human choriocarcinoma cells[J].Biochem Biophys Res Commun,2018503(4):3155-3161.
[13]Yoo YA,Kang MH,Lee HJ,et al.Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt,EMT,and MMP-9 pathway in gastric cancer[J].Cancer Res,2011,71(22)7061-7070.
[14]Esteva FJ,Guo H,Zhang S,et al.PTEN,PIK3CA,p-AKT,and p-p70S6K status:association with trastuzumab response and surviva in patients with HER2-positive metastatic breast cancer[J].Am JPathol,2010,177(4):1647-1656.
[15]Mabuchi S,Kuroda H,Takahashi R,et al.The PI3K/AKT/mTORpathway as a therapeutic target in ovarian cancer[J].Gynecol Oncol2015,137(1):173-179.
[16]Fumarola C,Bonelli MA,Petronini PG,et al.Targeting PI3K/AKT/mTOR pathway in non small cell lung cancer[J].Biochem Pharmacol,2014,90(3):197-207.
[17]Fresno Vara JA,Casado E,de Castro J,et al.PI3K/Akt signalling pathway and cancer[J].Cancer Treat Rev,2004,30(2):193-204.
[18]Silva BS,Yamamoto FP,Pontes FS,et al.T WIST and p-Akt immunoexpression in normal oral epithelium,oral dysplasia and in oral squamous cell carcinoma[J].Med Oral Patol Oral Cir Bucal2012,17(1):e29-34.
[19]Chang CH,Lee CY,Lu CC,et al.Resveratrol-induced autophagy and apoptosis in cisplatin-resistant human oral cancer CAR cells:Akey role of AMPK and Akt/mTOR signaling[J].Int J Oncol,201750(3):873-882.
[2]Lo WL,Kao SY,Chi LY,et al.Outcomes of oral squamous cel carcinoma in Taiwan after surgical therapy:factors affecting survival[J].J Oral Maxillofac Surg,2003,61(7):751-758.
[3]Kasala ER,Bodduluru LN,Madana RM,et al.Chemopreventive and therapeutic potential of chr ysin in cancer:mechanistic perspectives[J].Toxicol Lett,2015,233(2):214-225.
[4]Nile SH,Keum YS,Nile AS,et al.Antioxidant,anti-inflammatory and enzyme inhibitory activity of natural plant flavonoids and their synthesized derivatives[J].J Biochem Mol Toxicol,2018,32(1)e22002.
[5]Khoo BY,Chua SL,Balaram P.Apoptotic effects of chrysin in human cancer cell lines[J].Int J Mol Sci,2010,11(5):2188-2199.
[6]Wang J,Qiu J,Dong J,et al.Chr ysin protects mice from Staphylococcus aureus pneumonia[J].J Appl Microbiol,2011111(6):1551-1558.
[7]Du Q,Gu X,Cai J,et al.Chr ysin attenuates allergic air way inflammation by modulating the transcription factors T-bet and GATA-3 in mice[J].Mol Med Rep,2012,6(1):100-104.
[8]Chen YH,Yang ZS,Wen CC,et al.Evaluation of the structureactivity relationship of flavonoids as antioxidants and toxicants o zebrafish larvae[J].Food Chem,2012,134(2):717-724.
[9]Gresa-arribas N,Serratosa J,Saura J,et al.Inhibition of CCAAT/enhancer binding protein delta expression by chrysin in microglia cells results in anti-inflammatory and neuroprotective effects[J].JNeurochem,2010,115(2):526-536.
[10]Lopez J,Tait SW.Mitochondrial apoptosis:killing cancer using the enemy within[J].Br J Cancer,2015,112(6):957-962.
[11]Yoon TM,Kim SA,Lee DH,et al.EGR1 regulates radiation-induced apoptosis in head and neck squamous cell carcinoma[J].Oncol Rep2015,33(4):1717-1722.
[12]Park W,Park S,Lim W,et al.Chr ysin disrupts intracellular homeostasis through mitochondria-mediated cell death in human choriocarcinoma cells[J].Biochem Biophys Res Commun,2018503(4):3155-3161.
[13]Yoo YA,Kang MH,Lee HJ,et al.Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt,EMT,and MMP-9 pathway in gastric cancer[J].Cancer Res,2011,71(22)7061-7070.
[14]Esteva FJ,Guo H,Zhang S,et al.PTEN,PIK3CA,p-AKT,and p-p70S6K status:association with trastuzumab response and surviva in patients with HER2-positive metastatic breast cancer[J].Am JPathol,2010,177(4):1647-1656.
[15]Mabuchi S,Kuroda H,Takahashi R,et al.The PI3K/AKT/mTORpathway as a therapeutic target in ovarian cancer[J].Gynecol Oncol2015,137(1):173-179.
[16]Fumarola C,Bonelli MA,Petronini PG,et al.Targeting PI3K/AKT/mTOR pathway in non small cell lung cancer[J].Biochem Pharmacol,2014,90(3):197-207.
[17]Fresno Vara JA,Casado E,de Castro J,et al.PI3K/Akt signalling pathway and cancer[J].Cancer Treat Rev,2004,30(2):193-204.
[18]Silva BS,Yamamoto FP,Pontes FS,et al.T WIST and p-Akt immunoexpression in normal oral epithelium,oral dysplasia and in oral squamous cell carcinoma[J].Med Oral Patol Oral Cir Bucal2012,17(1):e29-34.
[19]Chang CH,Lee CY,Lu CC,et al.Resveratrol-induced autophagy and apoptosis in cisplatin-resistant human oral cancer CAR cells:Akey role of AMPK and Akt/mTOR signaling[J].Int J Oncol,201750(3):873-882.