灵芝多糖对人肝癌细胞HepG2增殖及体外迁移的影响
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摘要
目的体外检测灵芝多糖对人肝癌细胞株Hep G2迁移的影响,体内检测灵芝多糖对荷瘤小鼠的增殖抑制作用。方法 Transwell法检测灵芝多糖组(加灵芝多糖960μg/m L)与阴性对照组(加等体积培养液)的Hep G2细胞迁移数,ELISA法分别检测两组的血管内皮生长因子(VEGF)蛋白含量,取细胞密度为1×107个/m L的对数生长期Hep G2细胞,于BALB/c裸鼠左侧腋下行无菌皮下接种,建立肝癌移植瘤BALB/c裸鼠模型,并用灵芝多糖作用于人皮下Hep G2移植瘤模型。结果灵芝多糖组的Hep G2细胞迁移数较阴性对照组明显减弱,差异有统计学意义(P<0.05),灵芝多糖治疗组12、24 h的侵袭抑制率分别为31.7%、64.0%,与阴性对照组比较有统计学意义(P<0.05),并具有时间依赖性;灵芝多糖组与阴性对照组相比,VEGF蛋白含量显著下降,具有时间依赖性;灵芝多糖可体内抑制肝癌皮下移植瘤,1周给药、2周给药的抑瘤率分别为18.98%、37.33%,与阴性对照组比较差异均有统计学意义(P<0.05),并具有时间依赖性。结论灵芝多糖对Hep G2细胞的增殖、迁移有抑制作用,其迁移抑制通过下调VEGF蛋白含量而实现。
Objective To test the effect of ganoderma lucidum polysaccharides( GLP) on migration of Hep G2 liver cancer cells in vitro,as well as to detect the inhibiting effect on the cancer cells of tumor-bearing mice. Methods Transwell method was used to test the migration ability of Hep G2 cells in the GLP treatment group and Control Group.ELISA method was used to test the vascular endothelial growth factor( VEGF) protein of Hep G2 cells in both groups.Hep G2 cells( 107/m L) were implanted into the left oxterof BALB/c naked mice. The inhibition of tumor growth was used to test the proliferation of the cancer cells. Results Compared with control group,the invasion inhibition rates were 31. 7% and 64. 0% after treatment with GLP for 12 hours and 24 hours,respectively. The migration ability of Hep G2 cells in the treatment group was significantly reduced n a time-dependent manner( P < 0. 05). Compared with control group,the expression of VEGF protein in the treatment group was significantly reduced( P < 0. 05). GLP inhibited the growth of transplanted hepatocellular carcinoma on nude mice,while the anti-tumor rates were 18. 98% and 37. 33% after treatment with GLP for one week and two weeks,respectively; significantly differed from control group. Conclusion GLP can inhibit the proliferation and migration of Hep G2 cells by down-regulating VEGF proteins.
Objective To test the effect of ganoderma lucidum polysaccharides( GLP) on migration of Hep G2 liver cancer cells in vitro,as well as to detect the inhibiting effect on the cancer cells of tumor-bearing mice. Methods Transwell method was used to test the migration ability of Hep G2 cells in the GLP treatment group and Control Group.ELISA method was used to test the vascular endothelial growth factor( VEGF) protein of Hep G2 cells in both groups.Hep G2 cells( 107/m L) were implanted into the left oxterof BALB/c naked mice. The inhibition of tumor growth was used to test the proliferation of the cancer cells. Results Compared with control group,the invasion inhibition rates were 31. 7% and 64. 0% after treatment with GLP for 12 hours and 24 hours,respectively. The migration ability of Hep G2 cells in the treatment group was significantly reduced n a time-dependent manner( P < 0. 05). Compared with control group,the expression of VEGF protein in the treatment group was significantly reduced( P < 0. 05). GLP inhibited the growth of transplanted hepatocellular carcinoma on nude mice,while the anti-tumor rates were 18. 98% and 37. 33% after treatment with GLP for one week and two weeks,respectively; significantly differed from control group. Conclusion GLP can inhibit the proliferation and migration of Hep G2 cells by down-regulating VEGF proteins.
引文
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[14]孟崇.姜黄素对原发性肝癌模型大鼠组织VEGF及微血管密度影响研究[J].辽宁中医药大学学报,2017,19(2):19-22.
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[16]冯媽.灵芝多糖对DEN诱导的小鼠肝脏组织形态和原癌基因及抑癌基因的影响[D].晋中:山西农业大学,2013.
[17]Sun X,Zhao C,Pan W,et al.Carboxylate groups play a major role in antitumor activity of Ganoderma applanatum polysaccharide[J].Carbohydr Polym,2015,123:283-287.
[18]李杨,齐建利,赵立平,等.x CT影响肝癌细胞转移的作用机制研究[J].天津医科大学学报,2014,20(2):93-97.
[19]周森君.NEDD9促进肝细胞肝癌侵袭转移的作用研究DIXDC1在肝细胞肝癌中的表达及其与预后关性[D].杭州:浙江大学,2017.
[2]郭鹏荣,盛玉文,刘奔,等.灵芝多糖对顺铂抑制荷膀胱癌T24细胞裸鼠肿瘤生长及血管生成作用的影响[J].解放军医学杂志,2014,39(6):470-474.
[3]李红,陆洁,段昕所,等.灵芝多糖对小鼠黑素瘤B16F10细胞分泌VEGF的抑制作用[J].承德医学院学报,2017,34(4):276-278.
[4]林志彬.灵芝抗肿瘤作用的免疫学机制及其临床应用[J].中国药理学与毒理学杂志,2015,29(6):865-882.
[5]韦永福,陈晓颖,吴龙娟,等.灵芝多糖对肝癌细胞株Hep G2的PTEN活性的影响[J].右江民族医学院学报,2013,35(4):455-457.
[6]倪洪波.肝癌治疗的新进展[J].中国肿瘤临床与康复,2014,21(9):1148-1150.
[7]蒋亚波,郭卫星,程树群.肝细胞癌伴门静脉癌栓发生机制的研究进展[J].中国细胞生物学学报,2017,39(2):244-249.
[8]Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[9]Chen W,Zheng R,Baade PD,et al.Cancer stastistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[10]贺珊,廖长秀.中药治疗肝癌机制的研究进展[J].中成药,2017,39(1):155-160.
[11]张蕊,袁发璐,李婷,等.美洲大蠊提取物对人肝癌Hep G2细胞的作用机制研究[J].中国现代医学杂志,2017,27(12):1-8.
[12]徐铖,罗华荣,甘梅富.老年胃癌组织MMP-9、p53、VEGF蛋白的表达及其临床意义[J].中国老年学杂志,2017,37(10):2470-2472.
[13]张蕊,袁发璐,李婷,等.美洲大蠊提取物对人肝癌Hep G2细胞的作用机制研究[J].中国现代医学杂志,2017,27(12):1-8.
[14]孟崇.姜黄素对原发性肝癌模型大鼠组织VEGF及微血管密度影响研究[J].辽宁中医药大学学报,2017,19(2):19-22.
[15]吴先闯,杜钢军,郝海军,等.玉米须多糖对H22荷瘤小鼠的肿瘤抑制作用及其对小鼠免疫功能的影响[J].华西药学杂志,2015,30(1):26-29.
[16]冯媽.灵芝多糖对DEN诱导的小鼠肝脏组织形态和原癌基因及抑癌基因的影响[D].晋中:山西农业大学,2013.
[17]Sun X,Zhao C,Pan W,et al.Carboxylate groups play a major role in antitumor activity of Ganoderma applanatum polysaccharide[J].Carbohydr Polym,2015,123:283-287.
[18]李杨,齐建利,赵立平,等.x CT影响肝癌细胞转移的作用机制研究[J].天津医科大学学报,2014,20(2):93-97.
[19]周森君.NEDD9促进肝细胞肝癌侵袭转移的作用研究DIXDC1在肝细胞肝癌中的表达及其与预后关性[D].杭州:浙江大学,2017.