CAJ | 학술논문

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,7～8d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.
목적 탐토백려호순대인피부린상세포암(간칭린암)A431세포주라서이식류생장적영향.방법 취대수생장기A431세포주접충우Balb/c (nu/nu)라서좌액하조모,7～8d후장A431라서이식류모형안수궤수자표법분위공백조、음성대조조(생리록화납용액조)、배린선알(CTX)양성대조조、백려호순고、중、저제량조,매조10지.통과종말류질량계산백려호순적억류솔,관찰각조이식류조직병리형태학변화,원위말단표기기술검측세포조망,Western인적검측각조이식류조직중P-ERK、p53、반광안산천동안산단백매3(caspase3)적표체.SPSS13.0통계연건대결과진행방차분석급Pearson상관분석.결과 CTX조、백려호순고、중、저제량조、음성대조조、공백조라서이식류체적분별위(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,각조차이유통계학의의(F=16.00,P＜0.05)；각조류질량차이역유통계학의의(F=19.15,P＜ 0.05),백려호순고、중、저제량조억류솔분별위45.57％、37.97％、15.51％.CTX조、백려호순고、중、저제량조、음성대조조、공백조종류조직적조망지수분별위36.79±8.86、33.15±6.00、18.09±3.92、10.53±4.20、3.87±1.63、2.73±1.61,백려호순각조균고우음성대조조급공백조(F=93.26,P＜0.05).각조종류조직중P-ERK/β기동단백、p53/β기동단백caspase3/β기동단백비치경방차분석,F치분별위6.65、6.78、11.56,균P＜0.05,백려호순각조P-ERK、p53、caspase3단백적표체균고우음성대조조급공백조.경Pearson상관분석,라서이식류P-ERK여p53、p53여caspase3표체구유현저상관성(r분별0.68화0.56,균P＜0.05).결론 백려호순대A431하류라서이식류적생장구유억제작용,기궤제지일가능여활화종류조직중MAPK/ERK신호전도통로,진이활화p53,유도종류세포조망유관.
Objective To evaluate the effect of resveratrol on the growth of an established A431 xenogratt 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.