CXCR4下调对耐药腺样囊性癌细胞裸鼠移植瘤P-gp，VEGFR2及Ki-67表达的影响

发布时间：2018-06-01 21:05

本文选题：腺样囊性癌 + 免疫组化　；参考：《广西医科大学》2014年硕士论文

【摘要】：目的：研究靶向抑制CXCR4对耐药腺样囊性癌细胞系移植瘤中的P-gp，VEGFR2，Ki-67的表达，探讨CXCR4对耐药腺样囊性癌生物学特性的影响。方法：（1）在脂质体介导下，将靶向CXCR4基因的shRNA真核表达载体质粒体外转染腺样囊性癌耐药细胞系NACC-DDP，转染细胞建立裸鼠腮腺移植瘤，观察成瘤情况。（2）实验将裸鼠分为非转染组、空载体、阴性质粒对照组、shCXCR4组。接种后第35天处死动物，制备病理切片。 (3)通过组织切片脱蜡,水化,高压热修复,阻断过氧化物酶,滴加P-gp(1:200),VEGFR2(1:50),Ki-67等抗体,滴加生物素标记二抗工作液,滴加辣根酶标记的链霉卵白素工作液,DAB显色,苏木素复染,树胶封片等免疫组织化学法检测P-gp，VEGFR2，，Ki-67表达的情况。结果： 1. P-gp,VEGFR2主要表达于胞浆。分析结果显示，非转染组，空载体组，阴性质粒对照组P-gp,VEGFR2阳性表达率均高于shCXCR4组,差异有统计学意义(P0.05)。 2.Ki-67主要表达于细胞核。分析结果显示，非转染组，空载体组，阴性质粒对照组Ki-67阳性表达率高于shCXCR4组，差异有统计学意义(P0.05)。结论：下调CXCR4基因的NACC-DDP细胞移植瘤生长明显减慢，P-gp、VEGFR2、Ki-67表达明显降低，提示干扰CXCR4基因的表达可抑制耐药腺样囊性癌的增殖；下调耐药蛋白表达；并具有抗肿瘤血管生成的效应。CXCR4可望成为靶向抑制腺样囊性癌的靶点
[Abstract]:Objective: To investigate the effect of CXCR4 on the biological characteristics of drug-resistant adenoid cystic carcinoma (ADSCC), to investigate the inhibitory effect of CXCR4 on the expression of P-gptVEGFR2KI-67 in drug-resistant adenoid cystic carcinoma cell line transplantation and to investigate the effect of CXCR4 on the biological characteristics of drug-resistant adenoid cystic carcinoma. Methods: The adenoid cystic carcinoma resistant cell line NACC-DDP was transfected with shRNA eukaryotic expression vector plasmid targeting CXCR4 gene in vitro, and the xenograft tumor of parotid gland in nude mice was established by transfection of NACC-DDP. The nude mice were divided into three groups: non-transfection group, empty vector group and negative plasmid control group (shCXCR4). On the 35th day after inoculation, the animals were killed and pathological sections were prepared. (3) through tissue section dewaxing, hydration, high pressure thermal repair, blocking peroxidase, dripping with antibodies P-gp1: 1: 1: 1: 1: 50 + Ki-67, adding biotin-labeled second antibody working fluid, drip adding horseradish enzyme labeled streptavidin working fluid DAB, hematoxylin restaining, Immunohistochemical method was used to detect the expression of VEGFR2 Ki-67. Results: 1. VEGFR2 was mainly expressed in cytoplasm. The results showed that the positive expression rate of P-gpfR2 in non-transfection group, empty vector group and negative plasmid control group was higher than that in shCXCR4 group, and the difference was statistically significant (P 0.05). 2.Ki-67 is mainly expressed in the nucleus. The results showed that the positive expression rate of Ki-67 in non-transfection group, empty vector group and negative plasmid control group was higher than that in shCXCR4 group, and the difference was statistically significant (P 0.05). Conclusion: The growth of NACC-DDP cells with down-regulation of CXCR4 gene was significantly decreased, and the expression of P-gpCV-VEGFR2Ki-67 was significantly decreased, suggesting that interfering with the expression of CXCR4 gene could inhibit the proliferation of drug-resistant adenoid cystic carcinoma and down-regulate the expression of drug-resistant protein. CXCR4 is expected to be a target for the inhibition of adenoid cystic carcinoma.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.8

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