马勃多糖对乳腺癌荷瘤小鼠肿瘤生长的抑制作用及可能机制
|
摘要
为探讨马勃多糖(Calvatia gigantea polysaccharide, CGP)对乳腺癌荷瘤小鼠肿瘤生长的抑制作用及潜在机制,给小鼠接种4T1细胞建立乳腺癌荷瘤模型,将建模成功的小鼠随机分为模型组、环磷酰胺组(25 mg/kg)和CGP高(200 mg/kg)、中(100 mg/kg)、低(50 mg/kg)剂量组,每组8只;另随机选取8只健康小鼠为对照组(给予生理盐水)。灌胃给药,1次/d,给药周期为14 d。给药结束后检测瘤重、脾脏指数、胸腺指数,血清细胞因子水平及PI3K/Akt/mTOR信号通路相关蛋白表达。结果发现,与模型组比较,CGP高、中、低剂量组小鼠瘤重均明显减轻(P<0.05);与模型组比较,除CGP低剂量组小鼠胸腺指数外,各剂量组小鼠脾脏指数、胸腺指数、NK细胞增殖率、脾脏淋巴细胞增殖刺激指数及血清IFN-γ、IL-6、IL-2水平均明显增加(P<0.05);与模型组比较,CGP高、中、低剂量组小鼠p-PI3K、p-Akt和p-mTOR的相对表达量均明显降低(P<0.05)。以上结果提示CGP对乳腺癌荷瘤小鼠的肿瘤生长具有抑制作用,该作用与免疫调节及抑制PI3K/Akt/mTOR信号通路的激活有关。
The tumor growth inhibitory effect and mechanism of Calvatia gigantea polysaccharide(CGP) in mammary tumor-bearing mice were explored in this study. Mammary tumor-bearing mice were established by subcutaneous inoculation of 4 T1 cells on the back of the right side of the mice. The animals were then randomly divided into model group, cyclophosphamide group(25 mg/kg), and the high(200 mg/kg), medium(100 mg/kg) and low(50 mg/kg) dose CGP groups, with 8 mice each; in addition, 8 healthy mice were randomly selected as the normal group. The drugs were infused to the stomach of mice once a day for 14 days. Tumor weight, spleen index, thymus index, the activity of NK cells, spleen lymphocyte proliferation stimulation index, serum cytokine levels and protein expression of PI3 K/Akt/mTOR signaling pathway were detected, respectively. The results showed that, compared with the model group, the tumor weight was significantly decreased in the high, middle and low dose CGP groups(P < 0.05); moreover, in addition to the thymus index in the low dose CGP group, spleen index, thymus index, the activity of NK cells, spleen lymphocyte proliferation stimulation index and serum IFN-γ, IL-6, IL-2 levels were significantly increased in each dose CGP group(P < 0.05); furthermore, the relative expression levels of p-PI3 K, p-Akt and p-mTOR proteins were significantly decreased in all the 3 groups treated with CGP(P < 0.05). The above results suggest that CGP has tumor growth inhibitory effect on mammary tumor-bearing mice, and the mechanism may be related to the immunomodulatory effect and inhibition of the activation of PI3 K/Akt/mTOR signaling pathway.
The tumor growth inhibitory effect and mechanism of Calvatia gigantea polysaccharide(CGP) in mammary tumor-bearing mice were explored in this study. Mammary tumor-bearing mice were established by subcutaneous inoculation of 4 T1 cells on the back of the right side of the mice. The animals were then randomly divided into model group, cyclophosphamide group(25 mg/kg), and the high(200 mg/kg), medium(100 mg/kg) and low(50 mg/kg) dose CGP groups, with 8 mice each; in addition, 8 healthy mice were randomly selected as the normal group. The drugs were infused to the stomach of mice once a day for 14 days. Tumor weight, spleen index, thymus index, the activity of NK cells, spleen lymphocyte proliferation stimulation index, serum cytokine levels and protein expression of PI3 K/Akt/mTOR signaling pathway were detected, respectively. The results showed that, compared with the model group, the tumor weight was significantly decreased in the high, middle and low dose CGP groups(P < 0.05); moreover, in addition to the thymus index in the low dose CGP group, spleen index, thymus index, the activity of NK cells, spleen lymphocyte proliferation stimulation index and serum IFN-γ, IL-6, IL-2 levels were significantly increased in each dose CGP group(P < 0.05); furthermore, the relative expression levels of p-PI3 K, p-Akt and p-mTOR proteins were significantly decreased in all the 3 groups treated with CGP(P < 0.05). The above results suggest that CGP has tumor growth inhibitory effect on mammary tumor-bearing mice, and the mechanism may be related to the immunomodulatory effect and inhibition of the activation of PI3 K/Akt/mTOR signaling pathway.
引文
[1]Ero■lu C,Se■me M,Atmaca P,et al.Extract of Calvatia gigantea inhibits proliferation of A549human lung cancer cells[J].Cytotechnology,2016,68(5):2075-2081.
[2]Kivrak I,Kivrak S,Harmandar M.Bioactive compounds,chemical composition,and medicinal value of the giant puffball,calvatia gigantea(higher basidiomycetes),from Turkey[J].Int J Med Mushrooms,2016,18(2):97-107.
[3]Lam YW,Ng TB,Wang HX.Antiproliferative and antimitogenic activities in a peptide from puffball mushroom Calvatia caelata[J].Biochem Biophys Res Commun,2001,289(3):744-749.
[4]Ng TB,Lam YW,Wang H.Calcaelin,a new protein with translation-inhibiting,antiproliferative and antimitogenic activities from the mosaic puffball mushroom Calvatia caelata[J].Planta Med,2003,69(3):212-217.
[5]赵友生,王进平,宋爱荣,等.马勃多糖提取及体外抗肿瘤研究[J].中国现代应用药学,2012,29(7):574-578.
[6]姜明华,田易玲,徐凌川.马勃多糖对荷瘤模型小鼠抑瘤作用及生命延长率的影响[J].山东中医药大学学报,2014,38(2):157-158.
[7]黄凯,李志孝,邓永康,等.药用真菌马勃多糖的分离纯化及结构分析[J].华西药学杂志,2008,23(5):516-518.
[8]Anderson WF,Chu KC,Chang S,et al.Comparison of age-specific incidence rate patterns for different histopathologic types of breast carcinoma[J].Cancer Epidemiol Biomarkers Prev,2004,13(7):1128-1135.
[9]Millsop JW,Sharon VR,Petukhova T,et al.Chemotherapy reaction induced by ixabepilone,a microtubule stabilizing agent,mimicking extramammary Paget's disease in a patient with breast carcinoma[J].J Cutan Pathol,2016,43(12):1215-1219.
[10]Tsai M,Finkelstein M,Lillemoe T,et al.Pathologic predictors of treatment response to neo-adjuvant chemotherapy for invasive lobular breast carcinoma[J].Breast J,2017,23(5):607-609.
[11]Jayakumar T,Chiu CC,Wang SH,et al.Anti-cancer effects of CME-1,a novel polysaccharide,purified from the mycelia of Cordyceps sinensis against B16-F10melanoma cells[J].J Cancer Res Ther,2014,10(1):43-49.
[12]Manna DK,Maity P,Nandi AK,et al.Structural elucidation and immunostimulating property of a novel polysaccharide extracted from an edible mushroomLentinus fusipes[J].Carbohydr Polym,2017,157(10):1657-1665.
[13]Zhao X,Zhou D,Liu Y,et al.Ganoderma lucidumpolysaccharide inhibits prostate cancer cell migration via the protein arginine methyltransferase 6signaling pathway[J].Mol Med Rep,2018,17(1):147-157.
[14]侯超,胡志希.抗癌防移片抑制4T1乳腺癌血管生成的机制研究[J].广州中医药大学学报,2015,32(5):861-864,971.
[15]Xia Q,Geng F,Zhang FF,et al.Enhancement of fibroblast activation proteinα-based vaccines and adenovirus boost immunity by cyclophosphamide through inhibiting IL-10expression in 4T1tumor bearing mice[J].Vaccine,2016,34(38):4526-4535.
[16]程伟,邢东炜,张闽光.中药多糖调节肿瘤免疫应答研究进展[J].现代免疫学,2017,37(3):242-246.
[17]王瑶,吕海燕,郭佳,等.放射治疗荷肾癌小鼠肿瘤生长抑制与脾细胞及肿瘤组织IL-2、IFN-γ和IL-10的关系[J].第四军医大学学报,2009,30(20):2122-2124.
[18]孙昊鑫,朱金华,郭慧君,等.二冬膏对小鼠诱发性肺肿瘤发生及TNF-α、IL-6、IL-10、Foxp3的影响[J].中药药理与临床,2013,29(6):1-3.
[19]Huang S,Shen Y,Pham D,et al.IRF4modulates CD8+T cell sensitivity to IL-2family cytokines[J].Immunohorizons,2017,1(6):92-100.
[20]张星星,童佳兵,杨程,等.芪玉三龙汤对肺癌移植瘤小鼠PI3K/Akt/mTOR通路PI3K、Akt、mTOR表达的影响[J].安徽中医药大学学报,2016,35(1):73-77.
[21]Wang F,Tan WH,Liu W,et al.Effects of miR-214on cervical cancer cell proliferation,apoptosis and invasion via modulating PI3K/AKT/mTOR signal pathway[J].Eur Rev Med Pharmacol Sci,2018,22(7):1891-1898.
[22]陈明,王举涛,吴珍妮,等.半枝莲总黄酮通过PI3K/AKT/mTOR通路诱导肿瘤细胞自噬的体内实验研究[J].中国中药杂志,2017,42(7):1358-1364.
[2]Kivrak I,Kivrak S,Harmandar M.Bioactive compounds,chemical composition,and medicinal value of the giant puffball,calvatia gigantea(higher basidiomycetes),from Turkey[J].Int J Med Mushrooms,2016,18(2):97-107.
[3]Lam YW,Ng TB,Wang HX.Antiproliferative and antimitogenic activities in a peptide from puffball mushroom Calvatia caelata[J].Biochem Biophys Res Commun,2001,289(3):744-749.
[4]Ng TB,Lam YW,Wang H.Calcaelin,a new protein with translation-inhibiting,antiproliferative and antimitogenic activities from the mosaic puffball mushroom Calvatia caelata[J].Planta Med,2003,69(3):212-217.
[5]赵友生,王进平,宋爱荣,等.马勃多糖提取及体外抗肿瘤研究[J].中国现代应用药学,2012,29(7):574-578.
[6]姜明华,田易玲,徐凌川.马勃多糖对荷瘤模型小鼠抑瘤作用及生命延长率的影响[J].山东中医药大学学报,2014,38(2):157-158.
[7]黄凯,李志孝,邓永康,等.药用真菌马勃多糖的分离纯化及结构分析[J].华西药学杂志,2008,23(5):516-518.
[8]Anderson WF,Chu KC,Chang S,et al.Comparison of age-specific incidence rate patterns for different histopathologic types of breast carcinoma[J].Cancer Epidemiol Biomarkers Prev,2004,13(7):1128-1135.
[9]Millsop JW,Sharon VR,Petukhova T,et al.Chemotherapy reaction induced by ixabepilone,a microtubule stabilizing agent,mimicking extramammary Paget's disease in a patient with breast carcinoma[J].J Cutan Pathol,2016,43(12):1215-1219.
[10]Tsai M,Finkelstein M,Lillemoe T,et al.Pathologic predictors of treatment response to neo-adjuvant chemotherapy for invasive lobular breast carcinoma[J].Breast J,2017,23(5):607-609.
[11]Jayakumar T,Chiu CC,Wang SH,et al.Anti-cancer effects of CME-1,a novel polysaccharide,purified from the mycelia of Cordyceps sinensis against B16-F10melanoma cells[J].J Cancer Res Ther,2014,10(1):43-49.
[12]Manna DK,Maity P,Nandi AK,et al.Structural elucidation and immunostimulating property of a novel polysaccharide extracted from an edible mushroomLentinus fusipes[J].Carbohydr Polym,2017,157(10):1657-1665.
[13]Zhao X,Zhou D,Liu Y,et al.Ganoderma lucidumpolysaccharide inhibits prostate cancer cell migration via the protein arginine methyltransferase 6signaling pathway[J].Mol Med Rep,2018,17(1):147-157.
[14]侯超,胡志希.抗癌防移片抑制4T1乳腺癌血管生成的机制研究[J].广州中医药大学学报,2015,32(5):861-864,971.
[15]Xia Q,Geng F,Zhang FF,et al.Enhancement of fibroblast activation proteinα-based vaccines and adenovirus boost immunity by cyclophosphamide through inhibiting IL-10expression in 4T1tumor bearing mice[J].Vaccine,2016,34(38):4526-4535.
[16]程伟,邢东炜,张闽光.中药多糖调节肿瘤免疫应答研究进展[J].现代免疫学,2017,37(3):242-246.
[17]王瑶,吕海燕,郭佳,等.放射治疗荷肾癌小鼠肿瘤生长抑制与脾细胞及肿瘤组织IL-2、IFN-γ和IL-10的关系[J].第四军医大学学报,2009,30(20):2122-2124.
[18]孙昊鑫,朱金华,郭慧君,等.二冬膏对小鼠诱发性肺肿瘤发生及TNF-α、IL-6、IL-10、Foxp3的影响[J].中药药理与临床,2013,29(6):1-3.
[19]Huang S,Shen Y,Pham D,et al.IRF4modulates CD8+T cell sensitivity to IL-2family cytokines[J].Immunohorizons,2017,1(6):92-100.
[20]张星星,童佳兵,杨程,等.芪玉三龙汤对肺癌移植瘤小鼠PI3K/Akt/mTOR通路PI3K、Akt、mTOR表达的影响[J].安徽中医药大学学报,2016,35(1):73-77.
[21]Wang F,Tan WH,Liu W,et al.Effects of miR-214on cervical cancer cell proliferation,apoptosis and invasion via modulating PI3K/AKT/mTOR signal pathway[J].Eur Rev Med Pharmacol Sci,2018,22(7):1891-1898.
[22]陈明,王举涛,吴珍妮,等.半枝莲总黄酮通过PI3K/AKT/mTOR通路诱导肿瘤细胞自噬的体内实验研究[J].中国中药杂志,2017,42(7):1358-1364.