發(fā)布時(shí)間：2019-01-28 07:06

【摘要】：目的探討B(tài)7-H3對(duì)BALB/C-nu小鼠和C57BL/6小鼠前列腺癌移植瘤的影響及其可能的機(jī)制。 方法研究分兩部分進(jìn)行： 第一部分：利用基因芯片技術(shù)檢測(cè)綠色熒光蛋白（GFP）基因穩(wěn)定轉(zhuǎn)染的小鼠前列腺癌RM-1細(xì)胞株（RM-1/GFP）、B7-H3和GFP基因均穩(wěn)定轉(zhuǎn)染的RM-1細(xì)胞株（RM-1/B7-H3）的基因表達(dá)譜，通過芯片數(shù)據(jù)分析篩選出上述兩種細(xì)胞株之間的差異表達(dá)基因，進(jìn)一步挖掘其生物學(xué)信息，尋找B7-H3對(duì)小鼠前列腺癌皮下移植瘤影響的可能機(jī)制。 第二部分：將RM-1/B7-H3細(xì)胞懸液接種到BALB/C-nu小鼠（A組）和C57BL/6（B組）小鼠左側(cè)腹股溝皮下，RM-1/GFP細(xì)胞懸液接種到BALB/C-nu小鼠（A組）和C57BL/6（B組）小鼠右側(cè)腹股溝皮下，建立BALB/C-nu小鼠和C57BL/6小鼠前列腺癌皮下移植瘤模型。觀察并記錄腫瘤的生長情況，，繪制腫瘤生長曲線；進(jìn)一步用RT-PCR驗(yàn)證差異表達(dá)基因Bcl-2在C57BL/6小鼠腫瘤組織中mRNA的表達(dá)水平。 結(jié)果第一部分：RM-1/B7-H3細(xì)胞株和RM-1/GFP細(xì)胞株之間的基因表達(dá)譜差異明顯；與RM-1/GFP細(xì)胞相比，RM-1/B7-H3細(xì)胞中表達(dá)上調(diào)1.5倍及1.5倍以上的有309個(gè)基因；表達(dá)下調(diào)1.5倍及1.5倍以上的有127個(gè)基因。涉及凋亡，免疫應(yīng)答，程序性細(xì)胞死亡的調(diào)節(jié)，細(xì)胞粘附、侵襲、轉(zhuǎn)移和細(xì)胞周期等多種生物學(xué)過程，以及鈣離子信號(hào)通路、G蛋白偶聯(lián)受體介導(dǎo)的信號(hào)通路等多種信號(hào)通路。 第二部分：在第15天和第17天，A組左側(cè)移植瘤生長速度快于右側(cè)移植瘤，差異有統(tǒng)計(jì)學(xué)意義P0.05)；在第9天、第11天、第13天、第15天和第17天，B組左側(cè)移植瘤生長速度快于右側(cè)移植瘤，差異有統(tǒng)計(jì)學(xué)意義(P0.05)；第一部分基因芯片結(jié)果顯示與抑制細(xì)胞凋亡相關(guān)的表達(dá)上調(diào)2倍以上的基因有6個(gè)，選取其中的Bcl-2基因行逆轉(zhuǎn)錄聚合酶鏈反應(yīng)（RT-PCR）檢測(cè)，RT-PCR結(jié)果顯示RM-1/B7-H3腫瘤組織中Bcl-2的mRNA相對(duì)表達(dá)量明顯高于RM-1/GFP腫瘤組織，差異具有統(tǒng)計(jì)學(xué)意義(P0.05)，與基因芯片結(jié)果一致。 結(jié)論無論T細(xì)胞存在與否，B7-H3均能促進(jìn)小鼠前列腺癌RM-1細(xì)胞皮下移植瘤的生長，其機(jī)制可能與Bcl-2基因上調(diào)從而抑制RM-1/B7-H3細(xì)胞凋亡密切相關(guān)。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.25

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