B7-H3对小鼠前列腺癌皮下移植瘤的影响

发布时间：2019-01-28 07:06

【摘要】：目的探讨B7-H3对BALB/C-nu小鼠和C57BL/6小鼠前列腺癌移植瘤的影响及其可能的机制。方法研究分两部分进行：第一部分：利用基因芯片技术检测绿色荧光蛋白（GFP）基因稳定转染的小鼠前列腺癌RM-1细胞株（RM-1/GFP）、B7-H3和GFP基因均稳定转染的RM-1细胞株（RM-1/B7-H3）的基因表达谱，通过芯片数据分析筛选出上述两种细胞株之间的差异表达基因，进一步挖掘其生物学信息，寻找B7-H3对小鼠前列腺癌皮下移植瘤影响的可能机制。第二部分：将RM-1/B7-H3细胞悬液接种到BALB/C-nu小鼠（A组）和C57BL/6（B组）小鼠左侧腹股沟皮下，RM-1/GFP细胞悬液接种到BALB/C-nu小鼠（A组）和C57BL/6（B组）小鼠右侧腹股沟皮下，建立BALB/C-nu小鼠和C57BL/6小鼠前列腺癌皮下移植瘤模型。观察并记录肿瘤的生长情况，，绘制肿瘤生长曲线；进一步用RT-PCR验证差异表达基因Bcl-2在C57BL/6小鼠肿瘤组织中mRNA的表达水平。结果第一部分：RM-1/B7-H3细胞株和RM-1/GFP细胞株之间的基因表达谱差异明显；与RM-1/GFP细胞相比，RM-1/B7-H3细胞中表达上调1.5倍及1.5倍以上的有309个基因；表达下调1.5倍及1.5倍以上的有127个基因。涉及凋亡，免疫应答，程序性细胞死亡的调节，细胞粘附、侵袭、转移和细胞周期等多种生物学过程，以及钙离子信号通路、G蛋白偶联受体介导的信号通路等多种信号通路。第二部分：在第15天和第17天，A组左侧移植瘤生长速度快于右侧移植瘤，差异有统计学意义P0.05)；在第9天、第11天、第13天、第15天和第17天，B组左侧移植瘤生长速度快于右侧移植瘤，差异有统计学意义(P0.05)；第一部分基因芯片结果显示与抑制细胞凋亡相关的表达上调2倍以上的基因有6个，选取其中的Bcl-2基因行逆转录聚合酶链反应（RT-PCR）检测，RT-PCR结果显示RM-1/B7-H3肿瘤组织中Bcl-2的mRNA相对表达量明显高于RM-1/GFP肿瘤组织，差异具有统计学意义(P0.05)，与基因芯片结果一致。结论无论T细胞存在与否，B7-H3均能促进小鼠前列腺癌RM-1细胞皮下移植瘤的生长，其机制可能与Bcl-2基因上调从而抑制RM-1/B7-H3细胞凋亡密切相关。
[Abstract]:Objective to investigate the effect of B7-H3 on transplanted prostate cancer in BALB/C-nu mice and C57BL/6 mice and its possible mechanism. Methods the study was carried out in two parts: in the first part, the stable transfection of green fluorescent protein (GFP) gene into mouse prostate cancer RM-1 cell line (RM-1/GFP) was detected by gene chip technique. The gene expression profiles of RM-1 cell line (RM-1/B7-H3) transfected with B7-H3 and GFP genes were stable. The differentially expressed genes between the two cell lines were screened by microarray data analysis, and their biological information was further mined. To explore the possible mechanism of the effect of B7-H3 on subcutaneous transplanted tumor of prostate cancer in mice. The second part: the RM-1/B7-H3 cell suspension was inoculated into the left groin of BALB/C-nu mice (group A) and C57BL/6 mice (group B). RM-1/GFP cell suspension was inoculated into the right groin of BALB/C-nu mice (group A) and C57BL/6 mice (group B) to establish subcutaneous transplanted tumor models of prostate cancer in BALB/C-nu mice and C57BL/6 mice. The tumor growth was observed and recorded, the tumor growth curve was drawn, and the mRNA expression level of differentially expressed gene Bcl-2 in the tumor tissues of C57BL/6 mice was further verified by RT-PCR. Results in the first part, there were significant differences in gene expression profiles between RM-1/B7-H3 cell line and RM-1/GFP cell line. Compared with RM-1/GFP cells, there were 309 genes up-regulated by 1.5 times and more than 1.5 times in RM-1/B7-H3 cells, and 127 genes were down-regulated by 1.5 times and more than 1.5 times. It involves many biological processes, such as apoptosis, immune response, regulation of programmed cell death, cell adhesion, invasion, metastasis and cell cycle, as well as many signal pathways, such as calcium signaling pathway, G protein-coupled receptor mediated signal pathway, and so on. The second part: on the 15th and 17th day, the growth rate of the left transplanted tumor in group A was faster than that in the right transplantation tumor, the difference was statistically significant (P0.05). On day 9, day 11, day 13, day 15 and day 17, the growth rate of left transplanted tumor in group B was faster than that of right transplanted tumor (P0.05). The results of the first part of the gene chip showed that there were 6 genes related to the up-regulation of apoptosis. The Bcl-2 gene was selected for reverse transcriptase polymerase chain reaction (RT-PCR). RT-PCR results showed that the relative expression of Bcl-2 in RM-1/B7-H3 tumor tissue was significantly higher than that in RM-1/GFP tumor tissue, the difference was statistically significant (P0.05), which was consistent with the results of gene chip. Conclusion No matter whether T cells exist or not, B7-H3 can promote the growth of subcutaneous transplanted tumor of RM-1 cells of prostate cancer in mice. The mechanism may be related to the up-regulation of Bcl-2 gene and the inhibition of apoptosis of RM-1/B7-H3 cells.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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