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Antitumor effect and mechanism of Gecko on human esophageal carcinoma cell lines in vitro and xenografted sarcoma 180 in Kunming mice

机译:壁虎对昆明小鼠食管癌细胞和异种肉瘤180的抑制作用及其机制。

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摘要

AIM: To investigate the anti-tumor effect of Chinese medicine Gecko on human esophageal carcinoma cell lines and xenografted sarcoma 180 in Kunming mice and its mechanism.METHODS: The serum pharmacological method was used in vitro. The growth rates of the human esophageal carcinoma cells (EC9706 or EC1) were measured by a modified 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The transplanted tumor model of the mouse S180 sarcoma was established. Fifty mice were randomly divided into five groups (n = 10). Three Gecko groups were treated respectively with oral administration of Gecko powder at a daily dose of 13.5 g/kg, 9 g/kg, and 4.5 g/kg. The negative group (NS group) was treated with oral administration of an equal volume of saline and the positive group (CTX group) was treated with 100 mg/kg Cytoxan by intraperitoneal injection at the first day. After 2 wk of treatment, the anti-tumor activity was evaluated by tumor tissue weighing. The impact on immune organ was detected based on the thymus index, spleen index, phagocytic rate and phagocytic index. The protein expression of vascular endothelin growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected by immunohistochemistry. The cell apoptotic rate was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay.RESULTS: The A value in each group treated with Gecko after 72 h was reduced significantly in EC9706 and in EC1. The tumor weight in each group of Gecko was decreased significantly (1.087 ± 0.249 vs 2.167 ± 0.592; 1.021 ± 0.288 vs 2.167 ± 0.592; 1.234 ± 0.331 vs 2.167 ± 0.592; P < 0.01, respectively). However, the thymus index and Spleen index of mice in Gecko groups had no significant difference compared with the NS group. The immunoreactive score of VEGF and bFGF protein expression of each Gecko group by immunohistochemical staining were lowered significantly. The apoptosis index (AI) of each group was increased progressively with increase of dose of Gecko by TUNEL.CONCLUSION: Gecko has anti-tumor effects in vitro and in vivo; induction of tumor cell apoptosis and the down-regulation of protein expression of VEGF and bFGF may be contributed to anti-tumor effects of Gecko.
机译:目的:探讨中药壁虎对昆明小鼠食管癌细胞系和异种肉瘤180的抗肿瘤作用及其机制。方法:采用体外血清药理学方法。通过改良的3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)测定法测量人食道癌细胞(EC9706或EC1)的生长速率。建立了小鼠S180肉瘤的移植瘤模型。将五十只小鼠随机分为五组(n = 10)。三个壁虎组分别以每日剂量13.5 g / kg,9 g / kg和4.5 g / kg的口服壁虎粉进行治疗。阴性组(NS组)口服等量生理盐水治疗,阳性组(CTX组)在第一天腹膜内注射100 mg / kg细胞毒素。治疗2周后,通过肿瘤组织称重评估抗肿瘤活性。根据胸腺指数,脾脏指数,吞噬率和吞噬指数检测对免疫器官的影响。免疫组织化学检测血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)的蛋白表达。结果:在EC9706和EC1中,壁虎处理72 h后,各组的A值均显着降低,这是通过末端脱氧核苷酸转移酶(TdT)介导的dUTP缺口末端标记(TUNEL)法检测的。每组壁虎的肿瘤重量均显着降低(分别为1.087±0.249 vs 2.167±0.592; 1.021±0.288 vs 2.167±0.592; 1.234±0.331 vs 2.167±0.592; P <0.01)。然而,壁虎组小鼠的胸腺指数和脾脏指数与NS组相比无显着差异。免疫组化染色显示,各壁虎组VEGF和bFGF蛋白表达的免疫反应评分明显降低。结论:壁虎具有体外体内抗肿瘤作用。诱导肿瘤细胞凋亡以及下调VEGF和bFGF蛋白表达可能是 Gecko 的抗肿瘤作用。

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