中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (4): 248-252.

• 论著 • 上一篇    下一篇

MAPK/ERK信号转导通路在白藜芦醇抑制A431细胞株裸鼠移植瘤生长中的作用机制研究

郝玉琴1,黄维星2,3,宁晓洪2,3,冯红霞2,3,张国惠2,3,李衡贵2,3,郝春光2,3   

  1. 1. 内蒙古医科大学第三附属医院——内蒙古包钢医院
    2. 内蒙古医学院第三附属医院
    3.
  • 收稿日期:2012-04-25 修回日期:2012-07-01 出版日期:2013-04-15 发布日期:2013-04-01
  • 通讯作者: 郝玉琴 E-mail:haoyuqin0472@163.com
  • 基金资助:
    内蒙古自治区自然科学基金

Roles of MAPK/ERK signaling pathway in the growth inhibition of an established A431 xenograft tumor in nude mice by resveratrol

  • Received:2012-04-25 Revised:2012-07-01 Online:2013-04-15 Published:2013-04-01

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摘要： 目的 探讨白藜芦醇对人皮肤鳞状细胞癌（简称鳞癌）A431细胞株裸鼠移植瘤生长的影响。 方法　取对数生长期A431细胞株接种于Balb/c （nu/nu）裸鼠左腋下造模，7 ～ 8 d后将A431裸鼠移植瘤模型按随机数字表法分为空白组、阴性对照组（生理氯化钠溶液组）、环磷酰胺（CTX）阳性对照组、白藜芦醇高、中、低剂量组，每组10只。通过终末瘤质量计算白藜芦醇的抑瘤率，观察各组移植瘤组织病理形态学变化，原位末端标记技术检测细胞凋亡，Western印迹检测各组移植瘤组织中P-ERK、p53、半胱氨酸天冬氨酸蛋白酶3（caspase3）的表达。SPSS13.0统计软件对结果进行方差分析及Pearson相关分析。 结果　CTX组、白藜芦醇高、中、低剂量组、阴性对照组、空白组裸鼠移植瘤体积分别为（1153.56 ± 255.41）、（1001.69 ± 115.08）、（1206.80 ± 175.88）、（1342.28 ± 211.12）、（1642.34 ± 225.85）、（1564.32 ± 156.49） mm3，各组差异有统计学意义（F = 16.00，P < 0.05）；各组瘤质量差异亦有统计学意义（F = 19.15，P < 0.05），白藜芦醇高、中、低剂量组抑瘤率分别为45.57%、37.97%、15.51%。CTX组、白藜芦醇高、中、低剂量组、阴性对照组、空白组肿瘤组织的凋亡指数分别为36.79 ± 8.86、33.15 ± 6.00、18.09 ± 3.92、10.53 ± 4.20、3.87 ± 1.63、2.73 ± 1.61，白藜芦醇各组均高于阴性对照组及空白组（F = 93.26，P < 0.05）。各组肿瘤组织中P-ERK/β肌动蛋白、p53/β肌动蛋白caspase3/β肌动蛋白比值经方差分析，F值分别为6.65、6.78、11.56，均P < 0.05，白藜芦醇各组P-ERK、p53、caspase3蛋白的表达均高于阴性对照组及空白组。经Pearson相关分析，裸鼠移植瘤P-ERK 与p53、p53与caspase3表达具有显著相关性（r分别0.68和0.56，均P < 0.05）。 结论　白藜芦醇对A431荷瘤裸鼠移植瘤的生长具有抑制作用，其机制之一可能与活化肿瘤组织中MAPK/ERK信号传导通路，进而活化p53，诱导肿瘤细胞凋亡有关。 【关键词】 癌，鳞状细胞； 丝裂原激活蛋白激酶激酶类； 肿瘤抑制蛋白质p53； 细胞系，肿瘤； 白藜芦醇

关键词: 癌，鳞状细胞, 白藜芦醇, 细胞系，肿瘤, 丝裂原激活蛋白激酶激酶类, 肿瘤抑制蛋白p53

Abstract: HAO Yu-qin *, HUANG Wei-xing, NING Xiao-hong, FENG Hong-xia, ZHANG Guo-hui, LI Heng-gui, HAO Chun-guang. *Department of Dermatology, Third Affiliated Hospital of Inner Mongolian Medical College, Baotou 014010 , Inner Mongolia, China 【Abstract】 Objective To evaluate the effect of resveratrol on the growth of an established A431 xenograft tumor in nude mice. Methods The model of human skin squamous cell carcinoma was established by inoculating A431 cells in log-phase growth into the left axillary fossa of Balb/c （nu/nu） nude mice. After 7 － 8 days, 60 mice bearing human A431 skin squamous cell carcinoma xenografts were randomly and equally divided into 6 groups: blank control group receiving no treatment, negative control group treated with intraperitoneal sodium chloride physiological solution, positive control group treated with intraperitoneal cyclophosphamide, high-, medium- and low-dose resveratrol groups treated with intraperitoneal resveratrol of 40, 20 and 10 μg per gram body weight per day, respectively. Tumor size was measured at a 4-day interval during the treatment course. After 14-day treatment, the mice were sacrificed. Xenograft tumors were removed from these mice and subjected to weight measurement, pathological examination by hematoxylin and eosin （HE） staining and apoptosis detection by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL）. Western blot was conducted to quantify the protein expression of apoptosis-related factors, including phosphorylated extracellular signal-regulated protein kinase （p-ERK）, p53 and caspase 3. Data were processed by SPSS 13.0 software, and statistical analysis was carried out by analysis of variance and Pearson correlation analysis. Results By the end of treatment, the xenograft tumor volume was （1153.56 ± 255.41） mm3, （1001.69 ± 115.08） mm3, （1206.80 ± 175.88） mm3, （1342.28 ± 211.12） mm3, （1642.34 ± 225.85） mm3 and （1564.32 ± 156.49） mm3, and the weight was （1.84 ± 0.30） g, （1.72 ± 0.39） g, （1.96 ± 0.40） g, （2.67 ± 0.73） g, （3.16 ± 0.52） g, and （3.33 ± 0.59） g, respectively in the positive control group, high-, medium- and low-dose resveratrol group, negative control group and blank control group. Significant differences were observed in the xenograft tumor volume （F = 16.00, P < 0.05） and weight （F = 19.15, P < 0.05） among the 6 groups. According to the tumor weight, the growth of tumor was inhibited by 45.57%, 37.97% and 15.51% respectively in the high-, medium- and low-dose resveratrol groups. Increased apoptotic index was observed in the positive control group, high-, medium- and low-dose resveratrol groups compared with the negative control group and blank control group （36.79 ± 8.86, 33.15 ± 6.00, 18.09 ± 3.92 and 10.53 ± 4.20 vs. 3.87 ± 1.63 and 2.73 ± 1.61, F = 93.26, P < 0.05）. Analysis of variance showed that the protein expressions of p-ERK, p53 and caspase 3 were all higher in the three resveratrol groups than in the negative control group and blank control group （F = 6.65, 6.78, 11.56, respectively, all P < 0.05）. The protein expression of p53 was statistically correlated with p-ERK （r = 0.68, P < 0.05） and caspase 3（r = 0.56, P < 0.05）. Conclusions Resveratrol shows an inhibitory effect on the growth of human A431 skin squamous cell carcinoma xenografts in nude mice, likely by increasing p53 expression and inducing tumor cell apoptosis via the activation of MAPK/ERK pathway. 【Key words】 Carcinoma, squamous cell; Mitogen-activated protein kinase kinases; Tumor suppressor protein p53; Cell line, tumor; Resveratrol

Key words: Mitogen-activated protein kinase kinase, tumor suppressor P53 protein