人参果多糖对人舌鳞癌CAL27细胞增殖的抑制作用及其机制

doi:10.13481/j.1671-587x.20200207

摘要/Abstract

摘要： 目的：探讨人参果多糖对人舌鳞状细胞癌（简称舌鳞癌）CAL27细胞的抑制作用，阐明其潜在的作用机制。方法：采用水提醇沉法提取水溶性人参果多糖（WGBP），高效液相色谱法（HPLC）对其单糖组成成分进行分析。将人舌鳞癌CAL27细胞分为对照组和不同浓度（1、2和5 g·L^-1）WGBP组。继续培养24 h后，采用CCK-8法检测各组细胞增殖率，流式细胞术检测各组细胞周期和细胞凋亡率，Western blotting法检测各组细胞凋亡相关蛋白的表达量。结果：WGBP主要由半乳糖和半乳糖醛酸组成，而阿拉伯糖和葡萄糖次之，再次为鼠李糖和甘露糖。与对照组比较，1、2和5 g·L^-1WGBP组CAL27细胞增殖率明显降低（P<0.05或P<0.01），且呈浓度依赖性。流式细胞术检测，与对照组比较，2和5 g·L^-1WGBP组S期CAL27细胞百分比明显降低（P<0.05或P<0.01），G₂/M期细胞百分比明显升高（P<0.05或P<0.01），且呈浓度依赖性。与对照组比较，1、2和5 g·L^-1WGBP组CAL27细胞凋亡率明显升高（P<0.05），且呈浓度依赖性。Western blotting法检测，与对照组比较，1、2和5 g·L^-1WGBP组CAL27细胞中pro-Caspase-3和pro-PARP蛋白表达量明显升高。结论：WGBP能通过诱导G₂/M期阻滞和激活Caspase-3引起级联反应，诱导细胞凋亡，从而抑制人舌鳞癌CAL27细胞增殖。

关键词: 水溶性人参果多糖, 舌鳞状细胞癌, 细胞周期, 细胞凋亡, 半乳糖

Abstract: Objective: To investigate the inhibitory effect of polysaccharides from panax ginseng fruit in the human tongue squamous cell carcinoma CAL27 cells,and to elucidate its potential mechanism. Methods: Water soluble ginseng berry polysaccharide (WGBP) was extracted by water extraction and alcohol precipitation method, and its monosaccharide components were analyzed by high performance liquid chromatography (HPLC). The human tongue squamous cell carcinoma CAL27 cells were divided into control group and different concentrations (1,2 and 5 g·L^-1) of WGBP groups. After culture for 24 h, CCK-8 method was used to detect the proliferation rates of the CAL27 cells in various groups, flow cytometry was used to detect the cell cycle and apoptotic rates of the CAL27 cells in various groups,and Western blotting method was used to detect the expression amounts of apoptosis-related proteins in the CAL27 cells in various groups. Results: WGBP was mainly composed of galactose and galacturonic acid, followed by arabinose, glucose, rhamnose and mannose. Compared with control group, the proliferation rates of CAL27 cells in 1,2 and 5 g·L^-1 WGBP groups were increased (P<0.05 or P<0.01) in a concentration-dependent manner.The flow cytometry results showed that compared with control group, the percentages of CAL27 cells in S phase in 2 and 5 g·L^-1 WGBP groups were decreased (P<0.05 or P<0.01), and the percentages of CAL27 cells in G₂/M phase were increased (P<0.05 or P<0.01) in a concentration-dependent manner.The Western blotting results showed that compared with control group, the apoptotic rates of CAL27 cells in 1,2 and 5 g·L^-1 WGBP groups were increased (P<0.05) in a concentration-dependent manner. Compared with control group, the expression amounts of pro-Caspase-3 and pro-PARP proteins in the CAL27 cells in 1,2 and 5 g·L^-1 WGBP groups were increased. Conclusion: WGBP can induce the apoptosis and inhibit the proliferation of human tongue squamous cell carcinoma CAL27 cells by inducing the G₂/M phase arrest and Caspase-3 activation.

Key words: water-soluble ginseng berry polysaccharide, tongue squamous cell carcinoma, cell cycle, apoptosis, galactose

中图分类号:

R739.86

王会俞, 张天夫, 郝苗, 张楚, 王晓峰. 人参果多糖对人舌鳞癌CAL27细胞增殖的抑制作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(02): 248-253.

WANG Huiyu, ZHANG Tianfu, HAO Miao, ZHANG Chu, WANG Xiaofeng. Inhibitory effect of polysaccharides from panax ginseng fruit on proliferation of human lingual squamous cell carcinoma CAL27 cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 248-253.

参考文献

[1] ZHENG C M, GE M H, ZHANG S S, et al. Oral cavity cancer incidence and mortality in China, 2010[J].J Cancer Res Ther, 2015, 11(Suppl 2):C149-C154.
[2] SCHWAM Z G, SOSA J A, ROMAN S, et al. Complications and mortality following surgery for oral cavity cancer:analysis of 408 cases[J]. Laryngoscope, 2015, 125(8):1869-1873.
[3] HAN H Y, LEE H E, KIM H J, et al. Kochia scoparia induces apoptosis of oral cancer cells in vitro and in heterotopic tumors[J]. J Ethnopharmacol, 2016, 192:431-441.
[4] CARNEIRO-NETO J N, DE-MENEZES J D, MOURA L B, et al. Protocols for management of oral complications of chemotherapy and/or radiotherapy for oral cancer:Systematic review and meta-analysis current[J]. Med Oral Patol Oral Cir Bucal, 2017, 22(1):e15-e23.
[5] CAO J F, HOU D, LU J B, et al. Anti-tumor activity of exopolysaccharide from Rhizopus nigricans Ehrenb on S180 tumor-bearing mice[J]. Bioorg Med Chem Lett, 2016,26(8):2098-2104.
[6] MAO G H, ZHANG Z H, FEI F, et al. Effect of Grifola frondosa polysaccharide on anti-tumor activity in combination with 5-Fu in Heps-bearing mice[J]. Int J Biol Macromol, 2019, 121:930-935.
[7] ZHU L, CAO J F, CHEN G C, et al. Anti-tumor and immunomodulatory activities of an exopolysaccharide from Rhizopus nigricans on CT26 tumor-bearing mice[J]. Int Immunopharmacol, 2016, 36:218-224.
[8] LIU L, JIA J, ZENG G, et al. Studies on immunoregulatory and anti-tumor activities of a polysaccharide from Salvia miltiorrhiza Bunge[J]. Carbohydr Polym, 2013, 92(1):479-483.
[9] LEE D Y, PARK C W, LEE S J, et al. Immunostimulating and antimetastatic effects of polysaccharides purified from ginseng berry[J]. Am J Chin Med, 2019, 47(4):823-839.
[10] WAN J Y, HUANG W H, ZHENG W, et al. Multiple effects of ginseng berry polysaccharides:plasma cholesterol level reduction and enteric neoplasm prevention[J]. Am J Chin Med, 2017, 45(6):1293-1307.
[11] BAO L D, WANG Y, MA R L, et al. Apoptosis-inducing effects of lentinan on the proliferation of human bladder cancer T24 cells[J]. Pak J Pharm Sci, 2015, 28(5):1595-1600.
[12] SOHRETOGLU D, HUANG S L. Ganoderma lucidum polysaccharides as an anti-cancer agent[J]. Anticancer Agents Med Chem, 2018, 18(5):667-674.
[13] XIONG W, LI J, JIANG R, et al. Research on the effect of ginseng polysaccharide on apoptosis and cell cycle of human leukemia cell line K562 and its molecular mechanisms[J]. Exp Ther Med, 2017, 13(3):924-934.
[14] 施明毅,赵姝婷,温川飙,等.人参果产品开发研究现状[J].亚太传统医药,2018,14(1):59-61.
[15] KIM J, CHO S Y, KIM S H, et al. Effects of Korean ginseng berry on skin antipigmentation and antiaging via FoxO3a activation[J]. J Ginseng Res, 2017, 41(3):277-283.
[16] LI Z P, KIM H J, PARK M S, et al. Effects of fermented ginseng root and ginseng berry on obesity and lipid metabolism in mice fed a high-fat diet[J]. J Ginseng Res, 2018, 42(3):312-319.
[17] KIM M, YI Y S, KIM J, et al. Effect of polysaccharides from a Korean ginseng berry on the immunosenescence of aged mice[J]. J Ginseng Res, 2018, 42(4):447-454.
[18] 冯孟鑫.人参果多糖的分级分离纯化及理化性质与抗肿瘤活性初探[D].北京:北京中医药大学,2016.
[19] PU X, STORR S J, ZHANG YM, et al. Caspase-3 and caspase-8 expression in breast cancer:caspase-3 is associated with survival[J]. Apoptosis, 2017, 22(3):357-368.
[20] SAIRANEN T, SZEPESI R, KARJALAINEN-LINDSBERG M L, et al. Neuronal caspase-3 and PARP-1 correlate differentially with apoptosis and necrosis in ischemic human stroke[J]. Acta Neuropathol, 2009, 118(4):541-552.
[21] SHI C Y, MA Q H, REN M Y, et al. Antitumorpharmacological mechanism of the oral liquid of Poriacocos polysaccharide[J]. J Ethnopharmacol, 2017, 209:24-31.
[22] ZENG G, SHEN H, TANG G, et al. A polysaccharide from the alkaline extract of Glycyrrhiza inflata induces apoptosis of human oral cancer SCC-25 cells via mitochondrial pathway[J]. Tumour Biol, 2015, 36(9):6781-6788.
[23] ZHANG W F, YANG Y, LI X, et al. Angelica polysaccharides inhibit the growth and promote the apoptosis of U251 glioma cells in vitro and in vivo[J]. Phytomedicine, 2017, 33:21-27.
[24] 李珊珊,祝贺,祁玉丽,等.人参果多糖的分离纯化及体外抗氧化活性研究[J].食品工业科技,2018, 39(4):73-76.
[25] ZHANG M, TANG XL, WANG F, et al. Characterization of Lycium barbarum polysaccharide and its effect on human hepatoma cells[J]. Int J Biol Macromol, 2013, 61:270-275.
[26] 张培,郑晓萍,马玉玲,等.党参多糖单糖组成与其对HepG₂细胞毒活性的相关分析[J].中草药,2016, 47(15):2684-2692.
[27] ZHANG Q, LV X, WU T, et al. Composition of Lycium barbarum polysaccharides and their apoptosis-inducing effect on human hepatoma SMMC-7721 cells[J]. Food Nutr Res, 2015, 59:28696.