利用慢病毒表达系统研究TUBB3与CD44、CD133、PD-L1和p53表达量的关系  

Study on the Relationship Between TUBB3 and the Expression Levels of CD44,CD133,PD-L1,and p53 Using a Lentiviral Expression System

作　　者：谢诗韵 肖利红 杨雪莲 王郑 杨丽诗 XIE Shiyun;XIAO Lihong;YANG Xuelian;WANG Zheng;YANG Lishi(Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院,泸州646000

出　　处：《病毒学报》2024年第6期1297-1306,共10页Chinese Journal of Virology

摘　　要：探讨利用慢病毒表达系统研究微管蛋白β-III(Tubulin Beta 3,TUBB3)在小鼠髓母细胞瘤中的表达及其机制。纳入我院动物实验中心行手术切除的67例小鼠髓母细胞瘤组织及相应癌旁组织(距肿瘤组织边缘5cm以上)标本,通过免疫组化检测TUBB3、PD-L1及肿瘤干细胞标志物在小鼠髓母细胞瘤中的表达,并分析其相关性。CCK-8及TRANSWELL检测TUBB3对髓母细胞瘤细胞增殖和侵袭的影响。蛋白质免疫印迹(Western blotting)分析了p53表达对TUBB3表达的影响。结果显示,TUBB3表达阳性率为82.1%。Kaplan-Meier分析显示TUBB3高表达与髓母细胞瘤术后总生存率低有关。利用慢病毒表达系统敲低TUBB3的表达量能抑制Daoy和D341细胞增殖侵袭。小鼠髓母细胞瘤中TUBB3与CD44、CD133、PD-L1和p53表达呈正相关。Western blotting显示,p53基因敲低使TUBB3表达上调。本研究利用慢病毒表达系统发现,TUBB3与小鼠髓母细胞瘤的发生密切相关,是小鼠髓母细胞瘤的潜在有效生物标志物和治疗靶点。This study investigates the expression and underlying mechanism of Tubulin Beta 3(TUBB3)in mouse medulloblastoma using a lentiviral expression system.medulloblastoma tissues and corresponding adjacent non-tumor tissues(located more than 5 cm from the tumor margin)were collected from the animal research center of our hospital.Immunohistochemical analysis was performed to detect the expression of TUBB3,PD-L1,and tumor stem cell markers in the medulloblastoma samples,followed by correlation analysis.CCK-8 and Transwell assays were used to evaluate the effect of TUBB3 on the proliferation and invasion of medulloblastoma cells.Western blotting analysis was conducted to examine the influence of p53expression on TUBB3.The results showed a positive expression rate of TUBB3 at 82.1%.Kaplan Meier analysis indicated that high TUBB3 expression was associated with low overall survival rate in medulloblastoma post-surgery.Knockdown of TUBB3 expression using a lentiviral system significantly inhibited the proliferation and invasion of Daoy and D341 cell lines.Furthermore,a positive correlation was observed between the expression levels of TUBB3 and those of CD44,CD133,PD-L1,and p53 in mouse medulloblastoma.Western blotting results revealed that knockdown of p53 led to an upregulation of TUBB3 expression.These findings suggest that TUBB3 is closely associated with the development of mouse medulloblastoma and may serve as a potential biomarker and therapeutic target.

关 键 词：小鼠髓母细胞瘤 TUBB3 生物标志物 P53 肿瘤干细胞 PD-L1 

分 类 号：R373.9[医药卫生—病原生物学]