慢病毒介导SHCBP1低表达对黑色素瘤B16细胞增殖的影响  

The effect of lentivirus-mediated down-regulation of SHCBP1 on the proliferation of melanoma B16 cells

作　　者：孙志阳 冯光 雷静静 唐铖铖 孙晨皓 王令[1] 路宏朝[1] SUN Zhiyang;FENG Guang;LEI Jingjing;TANG Chengcheng;SUN Chenhao;WANG Ling;LU Hongzhao(School of Biological Science Technology and Engineering,Shaanxi University of Technology,Hanzhong 723001,China)

机构地区：[1]陕西理工大学生物科学与工程学院,汉中723000

出　　处：《生物学杂志》2022年第1期11-16,共6页Journal of Biology

基　　金：陕西省教育厅科技计划项目(No.20JK0570)。

摘　　要：构建Shcbp1慢病毒干扰载体,探究SHCBP1在黑色素瘤B16细胞中的生物学功能。通过设计靶向干扰小鼠Shcbp1基因的shRNA-1和shRNA-2以及对照序列shRNA-NC,连接至慢病毒骨架载体,利用HEK293T细胞包装慢病毒颗粒,侵染B16细胞,检测干扰效率,并通过CCK-8和流式细胞术检测细胞活力和细胞周期。利用Western Blot检测增殖相关基因的表达。结果显示:敲低Shcbp1显著抑制B16细胞增殖;流式细胞术检测结果发现,敲低Shcbp1将B16细胞阻滞在G1期;ERK1/2的磷酸化水平降低,P21的表达水平升高。结果表明SHCBP1可能通过ERK1/2信号通路抑制P21的转录,加速B16细胞从G1到S期进程。To construct Shcbp1 lentiviral interference vector to explore the biological function of SHCBP1 in melanoma B16 cells,first,the shRNA-1 and shRNA-2 that target to interfere with the mouse Shcbp1 gene and the control sequence shRNA-NC were designed and connected to the lentiviral backbone vector,HEK293 cells were used to package the lentiviral particles to infect B16 cells,the interfer-ence efficiency was tested and CCK-8 and flow cytometry were used to detect cell viability and cell cycle.Western Blot was used to de-tect the expression of proliferation-related genes.The results showed that knockdown of Shcbp1 significantly inhibited the proliferation of B16 cells;flow cytometry results showed that knockdown of Shcbp1 blocked B16 cells in the G1 phase;the phosphorylation level of ERK1/2 decreased and the expression level of p21 increased.These results indicate that SHCBP1 may inhibit the transcription of P21 through the ERK1/2 signaling pathway and accelerate the progress of B16 cells from G1 to S phase.

关 键 词：SHCBP1 细胞增殖 B16细胞 慢病毒干扰 

分 类 号：Q813[生物学—生物工程] R739.5[医药卫生—肿瘤]