CSF-1R过表达对鼻咽癌6-10B裸鼠移植瘤生长影响及其机制研究  

Effects and mechanism of overexpressing CSF-1R gene on growth of human nasopharyngeal carcinoma cell line 6-10B in nude mice

作　　者：陈泽南 蒿艳蓉[1] CHEN Ze-nan;HAO Yan-rong(Cancer Center , People's Hospital of Guangxi Zhuang Autonomous Region , Nanning 530021 ,P. R. China;Ruikang Clinical Faculty ,Guangxi University of Chinese Medicine , Nanning 530001 ,P. R. China)

机构地区：[1]广西壮族自治区人民医院肿瘤中心,广西南宁530021 [2]广西中医药大学瑞康临床医学院,广西南宁530001

出　　处：《中华肿瘤防治杂志》2019年第13期917-923,931,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81260348);广西研究生教育创新计划(2018053)

摘　　要：目的集落刺激因子-1受体(colony-stimulating factor-1 receptor,CSF-1R)在鼻咽癌放疗抵抗型患者组织中表达明显高于放疗敏感型患者,体外实验证实其通过PI3K/AKT途径可以促进鼻咽癌细胞6 10B的增殖、迁移和侵袭.本实验探讨CSF 1R对人鼻咽癌6 10B细胞系裸鼠移植瘤生长的影响,并分别从增殖、凋亡和自噬等方面研究其可能的机制.方法建立人鼻咽癌裸鼠异体移植瘤模型,体外利用慢病毒构建的CSF-1R过表达载体LV-CSF1R(16957-1)转染到人鼻咽癌6-10B细胞为实验组(5只),以空载体慢病毒转染6-10B细胞为阴性对照组(5只),不做处理6-10B细胞为空白对照组(5只).观察并记录裸鼠成瘤情况,剥离瘤体后测定肿瘤体积和质量.用HE染色、免疫组化法、实时荧光定量聚合酶链反应(quantitative real time polymerase chain reaction,qRT PCR)法和蛋白质印迹法等检测移植瘤中CSF-1R基因、增殖相关基因、凋亡相关基因及自噬相关基因的表达.结果 HE染色和免疫组织化学显示实验组过表达CSF-1R.实验组瘤体平均体积为(1 071.747±214.399) mm^3,大于阴性对照组(297.943±86.802) mm^3和空白对照组(96.152±40.749) mm^3,差异有统计学意义,F=72.119,P<0.001;实验组、阴性对照组和空白对照组瘤体平均质量分别为(2.406±0.348)、(0.432±0.156)和(0.17±0.111)g,差异有统计学意义,F-141.852,P<0.001.qRT PCR法检测显示,实验组CSF-1R mRNA相对表达量为6.161±0.319,高于阴性对照组0.119±0.100和空白组0.076±0.044,差异有统计学意义,F=970.046,P<0.001;实验组CSF-1R蛋白表达量为1.711±0.201,高于阴性对照组0.025±0.009和空白对照组0.023±0.009,差异有统计学意义,F=210.049,P=0.013.空白对照组、阴性对照组和实验组Cyclin D1表达量分别为0.748±0.163、0.778±0.150和1.253±0.123,F-11.269,P=0.009;实验组MMP2表达量为1.080±0.051,高于空白对照组0.067±0.012和阴性对照组0.422±0.056,F=404.710,P<0.001;空白对照组与阴性对照OBJECTIVE The expression of colony-stimulating factor-1 receptor(CSF-1R) was significantly higher in patients with radiotherapy-resistant nasopharyngeal carcinoma than in radiotherapy-sensitive patients. We had confirmed that PI3K/AKT pathway can promote the proliferation, migration and invasion of nasopharyngeal carcinoma 6-10B cells in vitro. This study was to investigate the effect of CSF-1R on the growth of human nasopharyngeal carcinoma 6-10B cell line xenografts in nude mice and to study its possible mechanisms on proliferation, apoptosis and autophagy. METHODS CSF-1R overexpression vector LV-CSF1R(16957-1) constructed by lentivirus was transfected into human nasopharyngeal carcinoma 6-10B cells by injected into the experimental group(5 rats), empty vector was transfected into 6-10B cells by injected into the negative group(5 rats) and 6-10B cells were injected into the blank control(5 rats). The nude mouse xenograft model of human nasopharyngeal carcinoma was established,and the tumor formation of nude mice was observed. Tumor volume and mass were determined after the tumor was removed. Detection of CSF-1R gene was via HE staining, immunohistochemistry ,quantitative real-time polymerase chain reaction(Qrt-PCR) and Western blotting. Detection of proliferation- related genes,apoptosis-related genes and autophagy-relate genes in transplanted tumors were done by using Western blotting. RESULTS HE staining and immunohistochemistry showed that the experimental group overexpressed CSF-1R. The average volume of the tumor in the experimental group was (1 071. 747±214. 399) mm^3, which was greater than the negative control group (297. 943±86. 802) mm^3 and the blank control group (96. 152±40. 749) mm^3. The difference was statistically significant (F= 72. 119,P<0. 001). The mean mass of the tumors in the experimemt group,negative control group and blank control group were (2. 406±0. 348),(0. 432±0. 156) and (0. 17±0. 111) g respectively,and the difference was statistically significant (F = 141. 852, P<0. 001).

关 键 词：集落刺激因子-1受体 鼻咽癌 增殖 凋亡 自噬 

分 类 号：R739.6[医药卫生—肿瘤]