|本期目录/Table of Contents|

[1]张 静,高 雅,李新宇,等.SUZ12过表达及敲减恶性外周神经鞘瘤稳定细胞株的建立及其意义[J].天津医科大学学报,2019,25(05):429-435.
 ZHANG Jing,GAO Ya,LI Xin-yu,et al.Establishment and significance of SUZ12 overexpression and knockdown of stable malignant peripheral nerve sheath tumor cell lines[J].Journal of Tianjin Medical University,2019,25(05):429-435.





Establishment and significance of SUZ12 overexpression and knockdown of stable malignant peripheral nerve sheath tumor cell lines
张 静1高 雅2李新宇2朱香熹3孙硕遥1肖 可4王 静2刘嘉岳2赵玉龙1李光明1朱 泽1
(1.天津医科大学病原生物学系,天津300070;2.天津医科大学基础医学院,天津300070;3. 遵义医科大学珠海校区临床医学系,珠海 519090;4.多伦多大学圣乔治校区,多伦多 ONM5S)
ZHANG Jing1GAO Ya2LI Xin-yu2ZHU Xiang-xi3SUN Shuo-yao1XIAO Ke4WANG Jing2LIU Jia-yue2ZHAO Yu-long1 LI Guang-ming1ZHU Ze1
(1. Department of Pathogen Biology,Tianjin Medical University,Tianjin 300070,China;2. School of Basic Medical Sciences, Tianjin Medical University,Tianjin 300070,China;3. Department of Clinical Medicine, Zhuhai Campus of Zunyi Medical University, Zhuhai 519090,China;4. University of Toronro-St.George Campus, Toronro ONM5S, Canada)
SUZ12 恶性外周神经鞘瘤CRISPR/Cas9慢病毒
SUZ12 MPNST CRISPR/Cas9 lentivirus
Objective: To establish the malignant peripheral nerve sheath tumor(MPNST) stable cell lines for over-expressed SUZ12 and knockdown SUZ12 and explore its significance. Methods: The full length SUZ12 gene was synthesized by PCR method,and three specific sgRNA interfere target sequences(sgRNA-1, sgRNA-2 and sgRNA-3)were designed and synthesized. Then to construct recombinant expression plasmids, 293T cells were transfected with these recombinant plasmids to package lentivirus, of which the titer were measured by fluorescence method. The optimal MOI for lentiviral infection and the optimal dose for purinomycin screening were determined, and ST8814 cells were transfected to construct the MPNST stable cell lines for overexpression and knockdown. The expression of green fluorescent protein in stably transfected strains was observed by fluorescence microscopy, and RT-qPCR was verified at the gene level. MTT assay was used to verify the proliferative activity of SUZ12 overexpression and knockdown groups. Results: LV-SUZ12 and LV-SUZ12-sgRNA were transduced into ST8814 cells and stably expressed green fluorescent protein. The results of RT-qPCR showed that the expression of SUZ12 overexpression stably increased(P<0.05), and the expression of CRISPR/Cas9 knockdown strain SUZ12 decreased significantly(P<0.05). MTT results show that overexpression of SUZ12 gene inhibits proliferation of MPNST cells. Conclusion: The stably transfected cell lines with overexpression and knockdown of SUZ12 were successfully constructed, and preliminary validation results indicate that overexpression of SUZ12 gene may indicate the growth of MPNST cells, providing experimental basis for further study on the biological function, mechanism and diagnosis and treatment of SUZ12 in MPNST.


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基金项目 国家自然科学基金资助项目(81672650)
作者简介 张静(1994-),女,硕士在读,研究方向:恶性外周神经鞘瘤发病与发展的分子机制;通信作者:朱泽,E-mail: zhuze@tijmu.edu.cn,李光明,E-mail:li-guangming@hotmail.com。
更新日期/Last Update: 2019-10-11