Generation of h MSH2-gene knockdown lung cancer cell model with specific si RNAs in NCI-H520 cell line
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- 英文论文题名：Generation of h MSH2-gene knockdown lung cancer cell model with specific si RNAs in NCI-H520 cell line
- 论文作者：Dai Yu Mei ; Lu Bi Chao
- 英文论文作者：Dai Yu Mei ; Lu Bi Chao
- 年：2016
- 作者机构：广州市妇女儿童医疗中心;北京协和医学院&中国医学科学院基础医学研究所;
- 英文论文关键词：Small RNA interference ; Human Mut S homolog 2 ; Vγ9δ2 T cells ; Lung cancer
- 会议召开时间：2016-11-04
- 会议录名称：第十一届全国免疫学学术大会壁报交流集
- 英文会议录名称：Proceedings of 2016 CSI Congress on Immunology
- 语种：英文
- 分类号：R734.2
- 学会代码：IGMD
- 会议名称：第十一届全国免疫学学术大会
- 会议地点：中国安徽合肥
- 主办单位：中国免疫学会
- 学会名称：中国免疫学会
- 页数：1
- 文件大小：1129k
- 原文格式：D
- 会议级别：全国
摘要
Objective:To generate an h MSH2-gene knockdown lung cancer cell model with small RNA interference in NCI-H520 cell line,so as to verify the underlying mechanisms in h MSH2-mediated recognition and cytolyisis of lung cancer cells by Vγ9δ2 T cells.Methods:h MSH2 specific si RNAs were designed,synthesized and transfected into NCI-H520 cells.Gene knockdown efficiency was measured by q RT-PCR,Westernblot,flow cytometery and confocal microscopy 48 or 72 h after si RNAs transfection.Results:Transfection efficiency was up to 93% 6 h after si RNAs treatment.48 or 72 h after specific si RNAs transfection,si RNA duplex I and II respectively resulted in 97%,88% and 98%,97% reduction in h MSH2 m RNA expression when compared to mock control.Reduced h MSH2 protein expression was confirmed by Western blot.Decreased surface expression of h MSH2 in si RNA-treated groups was revealed by flow cytometery and confocal microscopy.Conclusion:si RNA-h MSH2 gene knockdown lung cancer cell model was successfully generated in NCI-H520 cell line,providing a useful means to explore the recognition and cytolytic mechanism in Vγ9δ2 T cell-mediated anti-lung cancer innate immunity.
Objective:To generate an h MSH2-gene knockdown lung cancer cell model with small RNA interference in NCI-H520 cell line,so as to verify the underlying mechanisms in h MSH2-mediated recognition and cytolyisis of lung cancer cells by Vγ9δ2 T cells.Methods:h MSH2 specific si RNAs were designed,synthesized and transfected into NCI-H520 cells.Gene knockdown efficiency was measured by q RT-PCR,Westernblot,flow cytometery and confocal microscopy 48 or 72 h after si RNAs transfection.Results:Transfection efficiency was up to 93% 6 h after si RNAs treatment.48 or 72 h after specific si RNAs transfection,si RNA duplex I and II respectively resulted in 97%,88% and 98%,97% reduction in h MSH2 m RNA expression when compared to mock control.Reduced h MSH2 protein expression was confirmed by Western blot.Decreased surface expression of h MSH2 in si RNA-treated groups was revealed by flow cytometery and confocal microscopy.Conclusion:si RNA-h MSH2 gene knockdown lung cancer cell model was successfully generated in NCI-H520 cell line,providing a useful means to explore the recognition and cytolytic mechanism in Vγ9δ2 T cell-mediated anti-lung cancer innate immunity.
Objective:To generate an h MSH2-gene knockdown lung cancer cell model with small RNA interference in NCI-H520 cell line,so as to verify the underlying mechanisms in h MSH2-mediated recognition and cytolyisis of lung cancer cells by Vγ9δ2 T cells.Methods:h MSH2 specific si RNAs were designed,synthesized and transfected into NCI-H520 cells.Gene knockdown efficiency was measured by q RT-PCR,Westernblot,flow cytometery and confocal microscopy 48 or 72 h after si RNAs transfection.Results:Transfection efficiency was up to 93% 6 h after si RNAs treatment.48 or 72 h after specific si RNAs transfection,si RNA duplex I and II respectively resulted in 97%,88% and 98%,97% reduction in h MSH2 m RNA expression when compared to mock control.Reduced h MSH2 protein expression was confirmed by Western blot.Decreased surface expression of h MSH2 in si RNA-treated groups was revealed by flow cytometery and confocal microscopy.Conclusion:si RNA-h MSH2 gene knockdown lung cancer cell model was successfully generated in NCI-H520 cell line,providing a useful means to explore the recognition and cytolytic mechanism in Vγ9δ2 T cell-mediated anti-lung cancer innate immunity.
引文