CYB5R2基因真核表达载体的构建及在鼻咽癌中的功能研究

发布时间：2018-08-09 20:24

【摘要】：目的：建立稳定表达CYB5R2基因的鼻咽癌细胞株,观察CYB5R2基因对鼻咽癌细胞恶性肿瘤生物行为的影响,验证其在鼻咽癌中的抑癌特性,为寻找更多的更有效的鼻咽癌分子生物学标记物及治疗新靶点提供理论依据和奠定实验基础。方法：我们利用从CYB5R2高表达的正常睾丸组织cDNA中扩增的CDS序列,构建CYB5R2基因真核表达载体；将其转染入CYB5R2表达沉默的鼻咽癌细胞株,筛选鉴定出稳定表达CYN5R2的鼻咽癌细胞株；通过生长抑制实验,克隆形成实验、迁移运动试验、裸鼠体内成瘤试验,观察CYB5R2对鼻咽癌细胞的增殖、侵袭以及成瘤能力的影响。结果：成功构建CYB5R2真核表达载体并获得稳定表达CYB5R2的鼻咽癌细胞株HONE1和CNE2。相对于转染的空载体的鼻咽癌细胞株,转染CYB5R2的鼻咽癌细胞的增殖能力、侵袭能力及裸鼠体内成瘤能力明显降低,均具有统计学意义。结论：我们的研究首次证明CYB5R2在体内外均可抑制鼻咽癌细胞的恶性生物学行为,是一个鼻咽癌相关的潜在肿瘤抑制基因。
[Abstract]:Objective: to establish a nasopharyngeal carcinoma cell line stably expressing CYB5R2 gene and observe the effect of CYB5R2 gene on the biological behavior of nasopharyngeal carcinoma (NPC) cells. In order to find more effective molecular biological markers of nasopharyngeal carcinoma and new therapeutic targets to provide theoretical basis and lay the experimental foundation. Methods: we constructed the eukaryotic expression vector of CYB5R2 gene by using the CDS sequence amplified from cDNA of normal testicular tissue with high expression of CYB5R2, transfected it into NPC cell line with CYB5R2 expression silencing, and screened and identified the NPC cell line expressing CYN5R2 stably. The effects of CYB5R2 on the proliferation, invasion and tumorigenesis of nasopharyngeal carcinoma cells were observed by growth inhibition test, clone formation test, migration exercise test and tumorigenesis test in nude mice. Results: CYB5R2 eukaryotic expression vector was successfully constructed and NPC cell lines HONE1 and CNE2 expressing CYB5R2 stably were obtained. Compared with nasopharyngeal carcinoma cell line transfected with empty vector, the proliferation, invasion and tumorigenic ability of CYB5R2 transfected nasopharyngeal carcinoma cell line were significantly lower than that of empty vector transfected nasopharyngeal carcinoma cell line. Conclusion: our results suggest that CYB5R2 can inhibit the malignant biological behavior of nasopharyngeal carcinoma cells in vivo and in vitro, and is a potential tumor suppressor gene associated with nasopharyngeal carcinoma.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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