| 钟昌桃 吴 媛 周训钊 颜 魁 姚孟薇 宁思思 朱玉平 韦正波 谢 莹.慢病毒转染对鼻咽癌细胞 5-8F 增殖迁移的影响[J].,2015,15(5):825-828 |
| 慢病毒转染对鼻咽癌细胞 5-8F 增殖迁移的影响 |
| Effect of Lentivirus Transfection on the Proliferation and Migration ofNasopharyngeal Carcinoma Line 5-8F Cells |
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| DOI： |
| 中文关键词: 鼻咽癌 5-8F 细胞 慢病毒 转染 |
| 英文关键词: Nasopharyngeal carcinoma 5-8F cells Lentivirus Transfection |
| 基金项目:国家自然科学基金项目（ 81 160329； 81360406） |
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| 中文摘要: |
| 目的： 探讨慢病毒转染对鼻咽癌细胞株 5-8F 增殖、迁移的影响， 以验证慢病毒转染是否能有效的应用 于鼻咽癌细胞的增值
及迁移相关研究。 方法： 以红色荧光标记的慢病毒为 转染载体，选定不同 的 MOI 值转染鼻咽癌 5-8F 细胞株， 扩大培养后筛 选纯
化，流式细胞仪检测转染效率。 以最佳 MOI 值转染后的 5-8F（ RFP-5-8F）细胞为实验组，未转染的亲代 5-8F 为 空白对照组， 取对数
生长期未转染的亲代 5-8F 和红色荧光标记的慢病毒转染的 5-8F（ RFP-5-8F）细胞进行 MTT、划痕实验， 观察细胞镜下形态， 了 解
细胞转染前后生长曲线， 细胞迁移能力的变化。 结果： 流式细胞仪检测 5-8F 细胞慢病毒转染效率大于 95 %， 转染最佳 MOI 值为
30
，镜下荧光强度适中。 实验组与对照组比较，转染前后 5-8F 细胞光镜形态相似， 生长曲线一致，差异无统计学意义（ P=0.997）， 划
痕实验显示 5-8F 与 RFP-5-8F 细胞迁移能力一致， 差异无统计学意义（ P>0.05）。 结论： 慢病毒转染后鼻咽癌细胞能真实有效的的
反应原细胞的增值及迁移能力， 可以很好的应用 于鼻咽癌增殖及其转移机制的相关研究。 |
| 英文摘要: |
| Objective:To explore the effects of lentivirus transfection on the proliferation and migration of Nasopharyngeal carcinoma line 5-8F cells, and to verify whether lentivirus transfection can be effectively applied to the research for the proliferation and migration of Nasopharyngeal carcinoma cell line 5-8F cells.Methods:Different MOI values were selected to transfect Nasopharyngeal carcinoma line 5-8F cells with red fluorescence-labeled lentivirus as transfection vector. RFP-5-8F cells were cultured and then were purified after transfection. Transfection efficiency was determined by flow cytometry after transfection. RFP-5-8F cells transfected optimal
MOI values was considered as the experimental group, and the untransfected parental 5-8F cells for the control group. MTT and Wound
healing assays were used to detect the migration and proliferation of 5-8F and RFP-5-8F in logarithmic phase, and the morphology of
these cells were observed under microscope. The cell growth curve and migration ability was analyzed.Results:Transfection efficiency
of 5-8F cells were higher than 95 % by flow cytometry, and the best MOI was 30 with moderate endoscopic fluorescence intensity. Compared with that in the control group, 5-8F and RFP-5-8F cells in the experimental group were morphologically similar under microscope,
and the growth curve of both were consistent, without statistical significance (P=0.997). Wound healing assays showed a consistent 5-8F
and RFP-5-8F cell migration ability, without statistical significance (P>0.05).Conclusion:: NPC cells after lentivirus transfection could effectively maintain their ability of proliferation and migration and could be applied to related mechanical research on the proliferation and
metastasis of NPC. |
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