海带多糖对喉癌细胞增殖和凋亡影响的实验探索
|
摘要
目的通过海带多糖干预人喉癌Tu212细胞系,探索其对Tu212细胞增殖和凋亡的影响及其抗肿瘤活性机制。方法不同浓度的海带多糖(80、160、320μg/ml)及顺铂(1.8 mg/L)分别作用于喉癌Tu212细胞,采用MTT比色法检测24、48、72小时后的生长抑制率;流式细胞仪检测48小时后Tu212细胞各期的生长情况;荧光显微镜观察48小时后Tu212细胞的凋亡情况;Western Blot检测CyclinBl,Bcl-2蛋白的表达情况。结果随着海带多糖浓度的增加,其对喉癌Tu212细胞的抑制率也升高,相同浓度的海带多糖对喉癌Tu212细胞的抑制率随着时间的延长而升高;海带多糖干预Tu212细胞48小时后,其增殖被阻滞在G2/M期;干预48小时后,荧光显微镜观察可见典型细胞凋亡;干预Tu212细胞48小时后,CyclinBl,Bcl-2蛋白表达下降,差异均有统计学意义。结论海带多糖对喉癌Tu212细胞的生长有抑制作用,其机制为通过抑制增殖及促进凋亡来实现。
OBJECTIVE To investigate the effect and its mechanism of laminaria japonica polysaccharides(LJP) on human laryngeal carcinoma cell Tu212 growth. METHODS The laryngeal squamous cell Tu212 in logarithmic growth phase were treated by laminaria japonica polysaccharides(80,160, 320 μg/ml) and cisplatin(1.8 mg/L). The MTT assay was used to evaluate cell growth inhibition rate after 24 h, 48 h and 72 h. The flow cytometry with PI staining were used to estimate cell cycle distribution after 48 h. The fluorescent microscopy was used to estimate cell apoptosis after 48 h. The western blot was used to evaluate the expressions of Cyclin B1 and Bcl-2. RESULTS The cell proliferation was inhibited by cellular LJP in a dose and time dependent manner. Tu212 cells proliferation were stopped at the G2/M phase treated with LJP after 48 h. Cell apoptosis was observed clearly by fluorescent microscope after 48 h. The expressions of Cyclin B1 and Bcl-2 were suppressed significantly treated with LJP after 48 h. All of the differences were significant(P<0.05). CONCLUSION LJP has inhibitive effects on the Tu212 cells, this mechanism was probably through inhibition of the proliferation and promotion of the apoptosis.
OBJECTIVE To investigate the effect and its mechanism of laminaria japonica polysaccharides(LJP) on human laryngeal carcinoma cell Tu212 growth. METHODS The laryngeal squamous cell Tu212 in logarithmic growth phase were treated by laminaria japonica polysaccharides(80,160, 320 μg/ml) and cisplatin(1.8 mg/L). The MTT assay was used to evaluate cell growth inhibition rate after 24 h, 48 h and 72 h. The flow cytometry with PI staining were used to estimate cell cycle distribution after 48 h. The fluorescent microscopy was used to estimate cell apoptosis after 48 h. The western blot was used to evaluate the expressions of Cyclin B1 and Bcl-2. RESULTS The cell proliferation was inhibited by cellular LJP in a dose and time dependent manner. Tu212 cells proliferation were stopped at the G2/M phase treated with LJP after 48 h. Cell apoptosis was observed clearly by fluorescent microscope after 48 h. The expressions of Cyclin B1 and Bcl-2 were suppressed significantly treated with LJP after 48 h. All of the differences were significant(P<0.05). CONCLUSION LJP has inhibitive effects on the Tu212 cells, this mechanism was probably through inhibition of the proliferation and promotion of the apoptosis.
引文
[1]Liu M,Liu Y,Cao MJ,et al.Antibacterial activity and mechanisms of depolymerized fucoidans isolated from Laminaria japonica.Carbohydr Polym,2017,172:294-305.
[2]Pan A,Wang Y,Yuan JM,et al.High-sensitive C-reactive protein and risk of incident type 2 diabetes:a case-control study nested within the Singapore Chinese Health Study.BMC Endocr Disord,2017,17(1):8.
[3]李小婷,殷辰俞,孟徐莲.褐藻糖胶抑制人肝癌细胞HepG2增殖的机制.南京医科大学学报,2011,31(9):1261-1265.
[4]孙文忠,魏媛媛,徐志文.海带多糖和茶多酚对鼻咽癌细胞HONE1和CNE2增殖的影响.临床耳鼻咽喉头颈外科杂志,2013,28(8):425-428.
[5]Xu X,Kim JY,Oh YR,et al.Production of biodiesel from carbon sources of macroalgae,Laminaria japonica.Bioresour Technol,2014,169:455-461.
[6]Yang N,Wang Y,Hui L,et al.Silencing SOX2 Expression by RNA Interference Inhibits Proliferation,Invasion and Metastasis,and Induces Apoptosis through MAP4K4/JNK Signaling Pathway in Human Laryngeal Cancer TU212 Cells.J Histochem Cytochem,2015,63(9):721-733.
[7]Li J,Li H,Zhang Z,et al.The anti-cancerous activity of recombinant trichosanthin on prostate cancer cell PC3.Biol Res,2016,49:21.
[8]邓长江,朱希强,郭学平.海带多糖药理作用的研究进展.食品与药品,2006,8(4):30-32.
[9]周兰柱,王文忠,孙哲,等.喉癌前病变及喉癌中Livin表达的意义及其与Caspase-3表达的关系.中国耳鼻咽喉头颈外科,2016,23(3):173-174.
[10]黄志刚.喉癌基础研究现状和展望.中国耳鼻咽喉头颈外科,2010,17(2):57-58,76.
[11]Zhao D,Xu J,Xu X.Bioactivity of fucoidan extracted from Laminaria japonica using a novel procedure with high yield.Food Chem,2018,245:911-918.
[2]Pan A,Wang Y,Yuan JM,et al.High-sensitive C-reactive protein and risk of incident type 2 diabetes:a case-control study nested within the Singapore Chinese Health Study.BMC Endocr Disord,2017,17(1):8.
[3]李小婷,殷辰俞,孟徐莲.褐藻糖胶抑制人肝癌细胞HepG2增殖的机制.南京医科大学学报,2011,31(9):1261-1265.
[4]孙文忠,魏媛媛,徐志文.海带多糖和茶多酚对鼻咽癌细胞HONE1和CNE2增殖的影响.临床耳鼻咽喉头颈外科杂志,2013,28(8):425-428.
[5]Xu X,Kim JY,Oh YR,et al.Production of biodiesel from carbon sources of macroalgae,Laminaria japonica.Bioresour Technol,2014,169:455-461.
[6]Yang N,Wang Y,Hui L,et al.Silencing SOX2 Expression by RNA Interference Inhibits Proliferation,Invasion and Metastasis,and Induces Apoptosis through MAP4K4/JNK Signaling Pathway in Human Laryngeal Cancer TU212 Cells.J Histochem Cytochem,2015,63(9):721-733.
[7]Li J,Li H,Zhang Z,et al.The anti-cancerous activity of recombinant trichosanthin on prostate cancer cell PC3.Biol Res,2016,49:21.
[8]邓长江,朱希强,郭学平.海带多糖药理作用的研究进展.食品与药品,2006,8(4):30-32.
[9]周兰柱,王文忠,孙哲,等.喉癌前病变及喉癌中Livin表达的意义及其与Caspase-3表达的关系.中国耳鼻咽喉头颈外科,2016,23(3):173-174.
[10]黄志刚.喉癌基础研究现状和展望.中国耳鼻咽喉头颈外科,2010,17(2):57-58,76.
[11]Zhao D,Xu J,Xu X.Bioactivity of fucoidan extracted from Laminaria japonica using a novel procedure with high yield.Food Chem,2018,245:911-918.