木鳖子对羟基桂皮醛对小鼠黑素移植瘤生长的抑制作用及机制研究
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摘要
目的探讨木鳖子单体化合物对羟基桂皮醛(p-hydroxycinnamaldehyde,PHD)对黑素瘤B16细胞体内生长的抑制作用。方法应用MTS法检测PHD对B16细胞增殖的影响,倒置相差显微镜下观察形态变化。构建黑素瘤B16细胞小鼠皮下移植瘤模型,分为2组:治疗组小鼠ip 2 mg/kg PHD,模型组注射相同体积的生理盐水。观察2组小鼠成瘤情况,并测量肿瘤体积和质量;免疫组织化学法检测肿瘤组织中酪氨酸酶(Tyr)、S-100B、MMP-9、p-P38和p-ERK蛋白的表达变化;苏木精-伊红染色法(HE)分析小鼠肝和肺组织的形态学变化。结果 PHD对黑素瘤B16细胞具有明显的生长抑制作用(P<0.01)。20μmol/L PHD处理B16细胞48 h后,细胞数量减少,呈典型分化形态。PHD治疗组的肿瘤体积和质量均明显低于模型组(P<0.01)。与模型组相比,治疗组小鼠肿瘤组织中Tyr和p-P38蛋白的表达水平升高,MMP-9、S-100B和p-ERK蛋白表达水平则明显降低。治疗组小鼠肝和肺无明显组织形态学改变,模型组小鼠肺部有肿瘤转移。结论 PHD对小鼠体内黑素移植瘤的生长具有显著抑制作用。
Objective To investigate the inhibitory effect of p-hydroxylcinnamaldehyde(PHD) from Momordicae Semen seeds on growth of mouse melanoma B16 cells in vivo. Methods The inhibitory effect of PHD on growth of mouse melanoma B16 cells was measured by MTS method. Morphological changes of B16 cells were observed by phase contrast microscope. The xenograft tumor models of B16 cells in mice were established and divided into two groups: The mice in treatment group were treated with PHD(2 mg/kg) and in control group(treated with equal volume of normal saline). The growth of xenograft tumors in mice was observed and their sizes and weights were measured. The expression of Tyr, MMP-9, S-100 B, p-P38, and p-ERK in tumor tissues was detected by immunohistochemical method. Morphological changes of lung and liver tissues in mice were observed by HE staining. Results PHD had obvious inhibitory effect on the growth of B16 cells in vitro(P < 0.01). The morphological changes of B16 cells were typically differentiated after treated with 20 μmol/L PHD for 48 h. The average volume and weight of tumor tissues in mice of PHD treatment group were significantly decreased as compared with those of the control group(P < 0.01). Compared to the control group, the expression levels of Tyr and p-P38 in PHD treatment group were increased(P < 0.05), while the expression levels of MMP-9, S-100 B, and p-ERK were decreased(P < 0.05). No obvious morphological changes were found in liver and lung tissues of mice in PHD treatment group and the control group. However, lung tumor metastasis was found in control group mice. Conclusion PHD has inhibitory effect on the growth of xenograft tumor of mouse melanoma cells in mice.
Objective To investigate the inhibitory effect of p-hydroxylcinnamaldehyde(PHD) from Momordicae Semen seeds on growth of mouse melanoma B16 cells in vivo. Methods The inhibitory effect of PHD on growth of mouse melanoma B16 cells was measured by MTS method. Morphological changes of B16 cells were observed by phase contrast microscope. The xenograft tumor models of B16 cells in mice were established and divided into two groups: The mice in treatment group were treated with PHD(2 mg/kg) and in control group(treated with equal volume of normal saline). The growth of xenograft tumors in mice was observed and their sizes and weights were measured. The expression of Tyr, MMP-9, S-100 B, p-P38, and p-ERK in tumor tissues was detected by immunohistochemical method. Morphological changes of lung and liver tissues in mice were observed by HE staining. Results PHD had obvious inhibitory effect on the growth of B16 cells in vitro(P < 0.01). The morphological changes of B16 cells were typically differentiated after treated with 20 μmol/L PHD for 48 h. The average volume and weight of tumor tissues in mice of PHD treatment group were significantly decreased as compared with those of the control group(P < 0.01). Compared to the control group, the expression levels of Tyr and p-P38 in PHD treatment group were increased(P < 0.05), while the expression levels of MMP-9, S-100 B, and p-ERK were decreased(P < 0.05). No obvious morphological changes were found in liver and lung tissues of mice in PHD treatment group and the control group. However, lung tumor metastasis was found in control group mice. Conclusion PHD has inhibitory effect on the growth of xenograft tumor of mouse melanoma cells in mice.
引文
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[12]Jiang Z,Xu J,Long M,et al.2,3,5,4′-tetrahydroxystilbene-2-O-β-D-gluco-side(THSG)induces melanogenesis in B16 cells by MAP kinase activation and tyrosinase upregulation[J].Life Sci,2009,85(9/10):345-350.
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[14]Kruijff S,Hoekstra H J.The current status of S-100B as a biomarker in melanoma[J].Eur J Surg Oncol,2012,38(4):281-285.
[15]Selimovic D,Hassan M,Haikel Y,et al.Taxol-induced mitochondrial stress in melanoma cells is mediated by activation of c-Jun N-terminal kinase(JNK)and p38pathways via uncoupling protein 2[J].Cell Signal,2008,20(2):311-322.
[16]Krishna M,Narang H.The complexity of mitogenactivated protein kinases(MAPKs)made simple[J].Cell Mol Life Sci,2008,65(22):3525-3544.
[2]Balch C M,Gershenwald J E,Soon S J,et al.Final version of 2009 AJCC melanoma staging and classication[J].J Clin Oncol,2009,27(36):6199-6206.
[3]田红,肖桂芝,田苗,等.黑色素瘤治疗药物的研究进展[J].现代药物与临床,2015,30(7):890-896.
[4]王英凯,张明磊,张立波.人参三醇组皂甙诱导分化人肝癌细胞的研究[J].临床肝胆病杂志,2003,19(4):242-243.
[5]景永奎,韩锐.大豆甙元(S86019)与乳香有效成分Bc-4或阿糖胞昔对HL-60细胞分化的联合诱导[J].药学学报,1993,28(l):11-16.
[6]Wang N,Wang L W,Gou B D,et al.Realgar-induced differentiation is associated with MAPK pathways in HL-60 cells[J].Cell Biol Int 2008,32(12):1497-1505.
[7]赵连梅,韩丽娜,商晓辉,等.木鳖子醇提物对黑素瘤B16细胞增殖的抑制作用及其可能机制[J].中国肿瘤生物治疗杂志,2010,17(1):13-18.
[8]赵连梅,韩丽娜,耿艺曼,等.木鳖子醇提物诱导小鼠黑素瘤B16细胞分化的作用机制[J].中国肿瘤生物治疗杂志,2012,19(4):381-386.
[9]耿艺曼,赵连梅,朱秀丽,等.木鳖子对羟基桂皮醛对黑素瘤B16细胞分化的影响及其机制[J].中草药,2013,44(14):1951-1956.
[10]赵连梅,耿艺曼,孙士萍,等.木鳖子单体化合物对羟基桂皮醛诱导小鼠黑素瘤B16细胞的分化及其机制[J].中国肿瘤生物治疗杂志,2014,21(3):282-287.
[11]Zhao L M,Han L N,Ren F Z,et al.An ester extract of Cochinchina momordica seeds induces differentiation of melanoma B16F1 cells via MAPKs signaling[J].Asian Pac J Cancer Prev,2012,13(8):3795-3802.
[12]Jiang Z,Xu J,Long M,et al.2,3,5,4′-tetrahydroxystilbene-2-O-β-D-gluco-side(THSG)induces melanogenesis in B16 cells by MAP kinase activation and tyrosinase upregulation[J].Life Sci,2009,85(9/10):345-350.
[13]Wevers K P,Kruijff S,Speijers M J,et al.S-100B:a stronger prognostic biomarker than LDH in stage IIIB-C melanoma[J].Ann Surg Oncol,2013,20(8):2772-2779.
[14]Kruijff S,Hoekstra H J.The current status of S-100B as a biomarker in melanoma[J].Eur J Surg Oncol,2012,38(4):281-285.
[15]Selimovic D,Hassan M,Haikel Y,et al.Taxol-induced mitochondrial stress in melanoma cells is mediated by activation of c-Jun N-terminal kinase(JNK)and p38pathways via uncoupling protein 2[J].Cell Signal,2008,20(2):311-322.
[16]Krishna M,Narang H.The complexity of mitogenactivated protein kinases(MAPKs)made simple[J].Cell Mol Life Sci,2008,65(22):3525-3544.