目的 研究华蟾素口服液对人胃癌细胞 SGC-7901裸鼠原位移植瘤生长的抑制作用及其作用机制。方法 取雄性Balb/c 裸鼠,应用人胃癌SGC-7901细胞建立裸鼠皮下移植瘤,利用皮下移植瘤建立原位移植瘤模型,随机分为模型组,5-氟尿嘧啶组(阳性组),华蟾素口服液高、低(24、12 mL·kg-1)组,每组6只。阳性组隔日腹腔注射给药2周,华蟾素口服液组连续灌胃给药3周,末次给药后取出瘤体,测量瘤体体积、瘤重及抑瘤率,HE染色观察瘤体组织病理改变,末端脱氧核苷酸转移酶介导的dUTP 缺口末端标记测定法[terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling,TUNEL]测定瘤体细胞凋亡,免疫组化及Western blot法检测半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达,Western blot法检测Bax、Bcl-2、磷酸化磷脂酰肌醇-3-羟激酶/磷脂酰肌醇-3-羟激酶(p-PI3K/PI3K),p-Akt/Akt蛋白表达。结果 与模型组比较,华蟾素高、低剂量组可明显降低瘤体体积及重量;促进肿瘤细胞肿胀、变性及坏死,减少肿瘤间质内血管生成;促进肿瘤细胞凋亡;明显增加裂解的半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)、Bax蛋白表达,降低Bcl-2、p-PI3K和p-Akt蛋白表达水平。结论 华蟾素口服液对人胃癌裸鼠原位移植瘤有明显的抑制生长作用,其作用机制可能与其抑制PI3K/Akt信号通路从而促进肿瘤细胞凋亡有关。
Abstract
OBJECTIVE To study the inhibitory effect and mechanism of Huachangsu(HCS) oral solution on the growth of human gastric cancer cell line SGC-7901 orthotopic transplantation tumor in nude mice. METHODS The model of human gastric cancer cell line SGC-7901 orthotopic transplantation tumor in nude mice was established and randomly divided into four groups: model, 5-fluorouracil(5-FU, postive group), HCS oral solution 12 and 24 mL·kg-1 group. The positive group was intraperitoneally injected every other day for 2 weeks, and the HCS oral liquid group was given intragastric administration daily for 3 weeks. The tumor was removed after the last dose. Tumor volume, tumor weight and tumor inhibition rate were measured. Histopathological changes of tumor tissues were observed by HE staining, and apoptosis of tumor cells was determined by TUNEL method. Immunohistochemistry and Western blot were used to determine the protein expressions of caspase-3, Bax, Bcl-2, p-PI3K/PI3K and p-Akt /Akt. RESULTS Compared with model group, HCS oral solution could significantly decrease the tumor volume and weight, promote tumor cell swelling, degeneration and necrosis, reduce tumor interstitial angiogenesis and promote tumor cell apoptosis, increase cleaved caspase-3 and Bax protein expression, and decrease Bcl-2, p-PI3K, and p-Akt protein expression. CONCLUSION The HCS oral solution has obvious inhibitory effect on the growth of gastric orthotopic transplantation tumor in nude mice, which maybe related to inhibiting PI3K/Akt signaling pathway and thereby promoting the apoptosis of tumor cells.
关键词
华蟾素口服液 /
SGC-7901细胞株 /
原位移植瘤 /
凋亡 /
PI3K/Akt信号通路
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Key words
Huachangsu oral solution /
cell line SGC-7901 /
orthotopic transplantation tumor /
apoptosis /
PI3K/Akt signaling pathway
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中图分类号：
R965
R73-36+1
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参考文献
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脚注
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基金
安徽省科技重大专项资助(201903a07020031)
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