敲除鼻咽癌SUNE-1-6-10B细胞株中的KiSS-1基因对裸鼠肺转移的影响  

Effect of KiSS-1 Gene Knockout of Nasopharyngeal Carcinoma Cell Line SUNE-1-6-10B on Pulmonary Metastasis in Nude Mice

作　　者：张磊[1] 江浩[1] ZHANG Lei;JIANG Hao(Department of Radiation Oncology,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院放疗科,安徽蚌埠233000

出　　处：《沈阳医学院学报》2018年第6期491-494,498,共5页Journal of Shenyang Medical College

摘　　要：目的:探讨在鼻咽癌细胞株SUNE-1-6-10B中敲除KiSS-1基因对于裸鼠肺转移能力的影响。方法:(1)应用CRISPR/Cas9技术敲除鼻咽癌细胞株SUNE-1-6-10B中的KiSS-1基因,采用Surveyor法和Western blot技术检测是否成功获得低表达KiSS-1基因的鼻咽癌细胞株。(2)将KiSS-1基因敲除的鼻咽癌细胞株和正常鼻咽癌细胞株(0.2 ml,5×106/ml)分别注入40只裸鼠腋下,各20只;25 d后,观察裸鼠肺转移灶的情况。结果:(1)利用CRISPR/Cas9技术成功敲除了SUNE-1-6-10B细胞株中的KiSS-1基因,并获得了能够稳定低表达KiSS-1基因的鼻咽癌细胞株。(2) 25 d后,KiSS-1基因敲除组和KiSS-1基因未敲除组裸鼠腋下移植瘤的重量分别为(6.72±0.37) g和(6.80±0.38) g,差异无统计学意义(P>0.05);肺转移灶生成率分别为40%(8/20)和10%(2/20),差异有统计学意义(P<0.05)。结论:CRISPR/Cas9技术能够敲除鼻咽癌细胞株SUNE-1-6-10B中的KiSS-1基因;KiSS-1基因对于鼻咽癌细胞肺转移具有一定的抑制作用。Objective： To investigate the effect of KISS-1 gene knockout of nasopharyngeal carcinoma（NPC） cell line SUNE-1- 6-10B on pulmonary metastatic ability in nude mice. Methods： （1） KISS-1 gene was knocked out in SUNE-1-6-10B cells by using CRISPR/Cas9 technique. Surveyor and Western blot ,＊-ere used to detect the success of KISS-1 gene knockout. （2） The SUNE-1-6- 10B cells （0.2 ml, 5 × 10^6/ml）with or without KISS-1 gene knockout were injected into nude mice respectively. Lung metastasis in nude mice ws observed after 25 days. Results： （1） KISS-1 gene in SUNE-1-6-10B cells was successfully knocked out by CRISPR/ Cas9 technique. The NPC cell line with low expression of KISS-1 gene was obtained. （2） After transplanted for 25 days, the tumor weight in nude mice with KISS-1 gene knockout and without KISS-1 gene knockout were （6.72 ± 0.37）g and （6.80±0.38）g respectively. There was no significant difference （P〉0.05）. The rate of puhnonary metastasis was 40% （8/20） and 10% （2/20） respectively with significant difference （P〈0.05）. Conclusions： CRISPR/Cas9 technology can be used to knockout KISS-1 gene in NPC cell line SUNE-1-6-10B. KISS-1 gene plays a role in inhibiting puhnonary metastasis of NPC cells.

关 键 词：鼻咽癌 KISS-1基因 基因敲除 CRISPR/Cas9 

分 类 号：R739.6[医药卫生—肿瘤]