目的基因CXCL-1慢病毒表达系统的构建及在人正常肝细胞中的功能研究

发布时间：2018-02-16 14:11

本文关键词： 细胞趋化因子趋化因子生长调节基因正常肝细胞生长和迁移　出处：《广西医科大学学报》2014年04期 　论文类型：期刊论文

【摘要】：目的:探讨含目的基因趋化因子生长调节基因1(CXCL-1)慢病毒表达系统的构建及CXCL-1对7702正常肝细胞生长、增殖、迁移生物学功能的影响。方法:构建含目的基因CXCL-1的慢病毒表达系统,建立携带GFP荧光标记并过度表达CXCL-1的7702正常肝细胞系,Western blot方法检测CXCL-1在转染细胞中的表达,细胞单克隆形成实验,MTT实验,Transwell实验检测7702细胞表达CXCL-1后生长、增殖和迁移能力的变化情况。结果:1成功构建目的基因CXCL-1慢病毒表达系统,病毒滴度检测结果为2.00E+8TU/mL;2转染48h后经流式细胞仪检测,转染率达94.5%;3CXCL-1/7702组克隆形成率显著高于其余两对照组(P0.05);4MTT比色法结果显示:CXCL-1/7702组分别与空载GFP/7702组及空白对照7702组相比较,生长速度明显较快(P0.05);5Transwell实验显示:CXCL-1/7702组、GFP/7702组、7702组肝细胞系均无迁移能力。结论:成功构建含目的基因CXCL-1的慢病毒表达系统,CXCL-1对正常肝细胞具有明显的促细胞生长与增殖功能,但无促进正常肝细胞迁移的功能。
[Abstract]:Objective: to investigate the construction of lentivirus expression system containing chemokine growth regulator gene 1 (CXCL-1) and the effect of CXCL-1 on the growth, proliferation and migration of 7702 normal hepatocytes. Methods: a lentivirus expression system containing the target gene CXCL-1 was constructed. A normal liver cell line 7702 carrying GFP fluorescent labeling and overexpression of CXCL-1 was established to detect the expression of CXCL-1 in transfected cells by Western blot. Results the target gene CXCL-1 lentivirus expression system was successfully constructed. The results showed that the virus titer was 2.00E8TU / mLn-2 transfection for 48h, then detected by flow cytometry. The clone formation rate of group 1 / 7702 was significantly higher than that of the other two control groups (P0.05CL-1 / 7702). The results showed that the colony formation rate of the control group was significantly higher than that of the control group (7702) and the control group (7702), compared with the control group (7702) and the control group (7702). The growth rate was significantly faster than that of P0.05 / 5Transwell experiment. The results showed that the liver cell lines of GFP / 7702 group had no migration ability. Conclusion: the lentivirus expression system (CXCL-1) containing the target gene CXCL-1 can promote the growth and proliferation of normal hepatocytes, and CXCL-1 can promote the growth and proliferation of normal hepatocytes. However, it had no function to promote the migration of normal hepatocytes.
【作者单位】：广西医科大学附属肿瘤医院肝胆外科;广西医科大学附属肿瘤医院实验研究部;
【基金】：广西自然科学基金资助项目(No.2012jjAA40482)
【分类号】：R363

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