【摘要】：目的 檢測(cè)富含AT序列的特異結(jié)合蛋白1（specialAT-rich sequence binding protein1，SATB1）的mRNA在高成瘤高轉(zhuǎn)移鼻咽癌細(xì)胞株（SUNE-1亞株-5-8F）、只成瘤不轉(zhuǎn)移鼻咽癌細(xì)胞株（SUNE-1亞株-6-10B）、正常人永生化鼻咽上皮細(xì)胞株（NP69-SV40T）及SATB1蛋白在鼻咽癌（nasopharyngeal carcinoma，NPC）組織中的表達(dá)，探討SATB1的表達(dá)與NPC臨床病理因素的關(guān)系。 方法 采用熒光定量RT-PCR（FQ-RT-PCR）的方法檢測(cè)5-8F、6-10B及NP69-SV40T細(xì)胞中SATB1的mRNA表達(dá)水平；采用免疫組化法檢測(cè)72例原發(fā)性NPC和30例鼻咽黏膜慢性炎癥組織（chronic nasopharyngitis tissue，CNT）中SATB1蛋白的表達(dá)，并對(duì)檢測(cè)結(jié)果與NPC的臨床病理參數(shù)進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 1.成功建立實(shí)時(shí)熒光定量RT-PCR的檢測(cè)方法，擴(kuò)增效率高(＞90%)。SATB1mRNA在5-8F細(xì)胞中的表達(dá)較NP69-SV40T細(xì)胞中的表達(dá)稍低，兩者間差異無顯著性（P＞0.05），5-8F細(xì)胞的表達(dá)量為6-10B細(xì)胞的4.09倍，兩者間比較差異有顯著性（P＜0.05）。 2. SATB1染色陽性顆粒定位于細(xì)胞核。在NPC和CNT中SATB1蛋白的陽性表達(dá)率分別為52.8%（38/72）和13.3%（4/30），與CNT比較，，NPC中SATB1表達(dá)上調(diào)，差異有統(tǒng)計(jì)學(xué)意義（P＜0.05）。伴有淋巴結(jié)轉(zhuǎn)移和不伴淋巴結(jié)轉(zhuǎn)移的NPC組織中SATB1陽性表達(dá)率分別為64.3%（27/42）和36.7%（11/30），有遠(yuǎn)處轉(zhuǎn)移和無遠(yuǎn)處轉(zhuǎn)移的NPC組織的陽性表達(dá)率分別為88.9%（8/9）和47.6%（30/63），兩者比較，差異均有統(tǒng)計(jì)學(xué)意義（P＜0.05）。NPC原發(fā)灶中SATB1高表達(dá)與患者性別、年齡、T分級(jí)和臨床分期無關(guān)（P＞0.05），與N分級(jí)及M分級(jí)有關(guān)（P＜0.05）。 結(jié)論 1.5-8F細(xì)胞（高成瘤高轉(zhuǎn)移細(xì)胞株）中SATB1mRNA的表達(dá)水平顯著高于6-10B細(xì)胞（只成瘤不轉(zhuǎn)移細(xì)胞株）中SATB1mRNA的表達(dá)水平，可以說明SATB1mRNA的表達(dá)水平與鼻咽癌細(xì)胞株的侵襲轉(zhuǎn)移能力呈正相關(guān)。 2.NPC組織中存在著SATB1蛋白的高表達(dá)，且表達(dá)水平顯著高于CNT組織，提示SATB1在NPC的發(fā)生發(fā)展中可能起著重要作用。 3.SATB1蛋白在NPC組織中的表達(dá)水平與淋巴結(jié)轉(zhuǎn)移和遠(yuǎn)處轉(zhuǎn)移正相關(guān)，提示SATB1可能參與NPC的侵襲轉(zhuǎn)移。 目的 研究順鉑（DDP）及X射線對(duì)鼻咽癌5-8F細(xì)胞SATB1mRNA表達(dá)水平的影響，并探討其相關(guān)的意義。 方法 以噻唑藍(lán)（MTT）法檢測(cè)不同濃度DDP處理不同時(shí)間后5-8F細(xì)胞生存率的變化；用DDP及X射線干預(yù)5-8F細(xì)胞后，提取細(xì)胞總RNA，采用熒光定量RT-PCR方法檢測(cè)SATB1mRNA表達(dá)情況。 結(jié)果 MTT實(shí)驗(yàn)表明順鉑對(duì)鼻咽癌5-8F細(xì)胞的增殖抑制存在劑量時(shí)間效應(yīng)關(guān)系。順鉑及X射線處理鼻咽癌5-8F細(xì)胞后，SATB1mRNA水平均顯著增高（P＜0.05），差異有統(tǒng)計(jì)學(xué)意義。SATB1mRNA在鼻咽癌5-8F細(xì)胞中的表達(dá)變化與順鉑的作用濃度、作用時(shí)間及X射線之間無明顯的劑量時(shí)間效應(yīng)關(guān)系(P＞0.05。) 結(jié)論 鼻咽癌5-8F細(xì)胞經(jīng)順鉑及X射線作用后，SATB1mRNA表達(dá)發(fā)生上調(diào)，推測(cè)SATB1可能參與了鼻咽癌的化療耐藥和放療抵抗。
[Abstract]:Objective to detect the specific binding protein 1 (specialAT-rich sequence binding protein1,SATB1) mRNA rich in AT sequence in Gao Cheng high metastatic nasopharyngeal carcinoma cell line (SUNE-1 substrain-5-8F). The expression of NP69-SV40T and SATB1 protein in nasopharyngeal carcinoma (nasopharyngeal carcinoma,NPC) tissues was detected by tumorigenesis and no metastasis of nasopharyngeal carcinoma cell line (SUNE-1 substrain -6-10B), and the expression of SATB1 protein in nasopharyngeal epithelial cell line (NP69-SV40T) and normal human immortalized nasopharyngeal epithelial cell line (NP69-SV40T). To investigate the relationship between the expression of SATB1 and clinicopathological factors of NPC. Methods fluorescence quantitative RT-PCR (FQ-RT-PCR) was used to detect the mRNA expression of SATB1 in 5-8FN6-10B and NP69-SV40T cells. Immunohistochemical method was used to detect the expression of SATB1 protein in 72 cases of primary NPC and 30 cases of nasopharyngeal mucosa chronic inflammation (chronic nasopharyngitis tissue,CNT). The results and clinicopathological parameters of NPC were analyzed statistically. Result 1. The detection method of real-time fluorescent quantitative RT-PCR was successfully established. The amplification efficiency was higher than 90%. The expression of SATB1mRNA in 5-8F cells was slightly lower than that in NP69-SV40T cells, and there was no significant difference between them (P > 0. 05). The expression of 5-8F cells was 4.09 times higher than that of 6-10B cells (P < 0.05). 2. The positive granules of SATB1 staining were located in the nucleus. The positive expression rates of SATB1 protein in NPC and CNT were 52.8% (38 / 72) and 13.3% (4 / 30), respectively. Compared with CNT, the expression of SATB1 in NPC was up-regulated (P < 0. 05). The positive rates of SATB1 expression in NPC with and without lymph node metastasis were 64.3% (27 / 42) and 36.7% (11 / 30), respectively. The positive expression rates of NPC with and without distant metastasis were 88.9% (8 / 9) and 47.6% (30 / 63), respectively. The difference was statistically significant (P < 0. 05). The high expression of SATB1 was not correlated with sex, age, T grade and clinical stage (P > 0. 05), but correlated with N grade and M grade (P < 0. 05). Conclusion 1.The expression of SATB1mRNA in 5-8F cells (Gao Cheng high metastatic cell line) was significantly higher than that in 6-10B cell line (only tumorigenic non-metastatic cell line), and the expression level of SATB1mRNA in 6-10B cell line was significantly higher than that in 6-10B cell line. The expression of SATB1mRNA was positively correlated with the ability of invasion and metastasis of nasopharyngeal carcinoma cell line. There is a high expression of SATB1 protein in 2.NPC tissues, and the expression level is significantly higher than that in CNT tissues, suggesting that SATB1 may play an important role in the occurrence and development of NPC. The expression of 3.SATB1 protein in NPC was positively correlated with lymph node metastasis and distant metastasis, suggesting that SATB1 might be involved in the invasion and metastasis of NPC. Objective to investigate the effects of cisplatin (DDP) and X-ray on the expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma (NPC). Methods the survival rate of 5-8F cells was determined by thiazolyl (MTT) assay after different concentrations of DDP. After 5-8F cells were treated with DDP and X-ray, total RNA, was extracted and the expression of SATB1mRNA was detected by fluorescence quantitative RT-PCR. Results MTT assay showed that cisplatin inhibited the proliferation of 5-8 F cells in a dose-time dependent manner. After treated with cisplatin and X-ray, the level of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma increased significantly (P < 0. 05), the difference was statistically significant. The expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma and the action concentration of cisplatin were significantly increased (P < 0. 05). There was no significant dose-time effect relationship between the time of action and X-ray (P > 0.05). Conclusion the expression of SATB1mRNA in 5-8F cells of nasopharyngeal carcinoma was up-regulated by cisplatin and X-ray, suggesting that SATB1 might be involved in chemotherapeutic resistance and radiotherapy resistance of nasopharyngeal carcinoma.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R739.63

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