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血管内皮生长因子的体内短发夹RNA干扰对耐药舌癌细胞的作用

[In vivo short hairpin RNA interference of vascular endothelial growth factor on drug-resistant tongue cancer cells].

作者信息

Nong Xiao-Lin, Li Hao, Xia Yong, Li Yan, Li Jia-Quan, Yang Yi-Ping

机构信息

Department of Oral and Maxillofacial Surgery, College of Stomatology, Guangxi Medical University, Nanning 530021, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2011 Jan;46(1):15-9.

Abstract

OBJECTIVE

To study the in vivo interference effects of vascular endothelial growth factor (VEGF) short hairpin RNA (shRNA) on xenografts of drug-resistant tongue cancer cells.

METHODS

Drug-resistant tongue caner cells Tca/Cisplatin (DDP) were injected subcutaneously into nude mice to establish xenograft models, which were randomly divided into non-transfected group, mock control group, control group transfected with scrambled sequence plasmid, interference group transfected with VEGF-shRNA expression plasmid. Liposome-mediated plasmid transfection was done in the latter three groups every three days. Xenografts were observed and tumor growth curve was measured. After the 10th transfection, tumors were anatomized and weigh. Microvessel density was detected by immunohistochemical staining. In situ hybridization assay was used to test VEGF mRNA, and immunohistochemistry to test VEGF, P-glycoprotein (P-gp), B cell lymphoma/leukemia-2 (bcl-2) and extracellular signal-regultaed kinase 2 (ERK-2) protein.

RESULTS

Tumor growth in VEGF-shRNA interference group was significantly slow. Tumor weight was (0.4781 ± 0.0860) g, microvessel density (7.35 ± 1.31)/view, VEGF mRNA (0.0767 ± 0.0234), VEGF protein (0.1301 ± 0.0433), P-gp (0.1517 ± 0.0184), bcl-2 (0.1218 ± 0.0251) and ERK-2 protein (0.1178 ± 0.0291) in VEGF-shRNA interference group; all of them were less than those in the other three groups (P < 0.05).

CONCLUSIONS

Inhibition targeting VEGF may become a potential therapy for drug-resistant tongue cancer.

摘要

目的

研究血管内皮生长因子(VEGF)短发夹RNA(shRNA)对耐药舌癌细胞异种移植瘤的体内干扰作用。

方法

将耐药舌癌细胞Tca/顺铂(DDP)皮下注射到裸鼠体内建立异种移植模型,随机分为未转染组、空载体对照组、转染乱序序列质粒的对照组、转染VEGF-shRNA表达质粒的干扰组。后三组每三天进行一次脂质体介导的质粒转染。观察异种移植瘤并测量肿瘤生长曲线。第10次转染后,解剖肿瘤并称重。通过免疫组织化学染色检测微血管密度。采用原位杂交法检测VEGF mRNA,免疫组织化学法检测VEGF、P-糖蛋白(P-gp)、B细胞淋巴瘤/白血病-2(bcl-2)和细胞外信号调节激酶2(ERK-2)蛋白。

结果

VEGF-shRNA干扰组肿瘤生长明显缓慢。VEGF-shRNA干扰组肿瘤重量为(0.4781±0.0860)g,微血管密度为(7.35±1.31)/视野,VEGF mRNA为(0.0767±0.0234),VEGF蛋白为(0.1301±0.0433),P-gp为(0.1517±0.0184),bcl-2为(0.1218±0.0251),ERK-2蛋白为(0.1178±0.0291);均低于其他三组(P<0.05)。

结论

靶向VEGF的抑制作用可能成为耐药舌癌的一种潜在治疗方法。

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