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目的 :探讨p16基因对乳腺癌的治疗作用 ,为乳腺癌 p16的基因治疗提供实验依据。 方法 :构建p16的逆转录病毒载体 pLMSN ,包装成病毒后 ,体外转导p16基因纯合性缺失的乳腺癌细胞株MCF 7,Westernblot证明该基因在转导后的MCF 7细胞中有表达 ,用活细胞计数、流式细胞仪、形态观察等方法来检测 p16表达在体外对MCF 7细胞生物学行为的影响。进行SCID鼠体内成瘤实验和逆转录病毒直接注射SCID鼠乳腺癌模型的治疗性实验来观察 p16对乳腺癌细胞的体内抑制作用。 结果 :外源性 p16在MCF 7细胞中表达后 ,体外细胞发生形态改变 ,生长明显慢于对照细胞 ,流式细胞计数显示G1期细胞增多 ,细胞在SCID鼠体内成瘤性下降 ,p16逆转录病毒直接注射有使SCID鼠乳腺肿瘤缩小的趋势。结论 :p16逆转录病毒对p16基因纯合性缺失的乳腺癌有一定治疗意义
Objective: To explore the therapeutic effect of p16 gene on breast cancer and provide experimental basis for p16 gene therapy in breast cancer. METHODS: The p16 retroviral vector pLMSN was constructed. After packaging the virus, the human breast cancer cell line homozygous for p16 gene was transduced in vitro. Western Blot demonstrated that the gene was expressed in transduced MCF 7 cells. The effect of p16 expression on the biological behavior of MCF 7 cells in vitro was detected by viable cell counting, flow cytometry, and morphological observations. The in vivo inhibitory effect of p16 on breast cancer cells was observed by performing tumorigenicity experiments in SCID mice and in a retroviral direct injection SCID mouse breast cancer model. RESULTS: After exogenous p16 expression in MCF 7 cells, the morphological changes of cells in vitro were observed, and the growth was significantly slower than that of the control cells. Flow cytometry showed that the number of cells in G1 phase increased, and the tumorigenicity of cells in SCID mice decreased, p16 reverse transcription. The direct injection of virus has a tendency to shrink SCID murine mammary tumors. Conclusion : p16 retrovirus has certain therapeutic value for the homozygous deletion of p16 gene in breast cancer