【摘要】：鼻咽癌是一個多因素遺傳性的頭頸部常見腫瘤，其發(fā)生發(fā)展具有多階段性、隱蔽性等特點，是我國常見的惡性腫瘤之一；高發(fā)區(qū)主要集中在南方五省(廣東、廣西、湖南、福建、江西)，有廣東癌之稱。盡管由于外科手術(shù)以及化療手段地進步，，但由于其發(fā)病部位的隱蔽以及檢測指標(biāo)的有效性等問題，導(dǎo)致其診斷、治療仍存在相當(dāng)大的困難，多數(shù)患者發(fā)現(xiàn)病癥進行診斷后已是癌癥后期；因此，急切需要一種針對于鼻咽癌的比較合理、有效的診斷以及預(yù)后手段。 Hedgehog（Hh）信號通路發(fā)現(xiàn)于果蠅中的一個進化保守的信號通路，最初的功能是調(diào)節(jié)胚胎發(fā)育，隨后發(fā)現(xiàn)其在人類的多種癌癥中異常激活，參與腫瘤的發(fā)生、分化、侵襲和轉(zhuǎn)移。然而，對于Hh信號通路是否在鼻咽癌中存在異常表達以及其在鼻咽癌中的功能性影響，目前并不明確。 本研究首先運用熒光定量PCR檢測了Hh信號通路的PTCH-1、SMO和GLI-1基因在鼻咽癌以及鼻咽炎臨床組織中的表達情況。研究顯示PTCH-1、SMO和GLI-1基因表達于全部的鼻咽組織中，而熒光定量PCR結(jié)果表明PTCH、GLI-1在鼻咽癌組織以及鼻咽炎組織中存在表達差異（P0.05），PTCH表達下調(diào)，GLI-1則表達上調(diào)，而SMO基因表達無差異。免疫組織化學(xué)檢測進一步顯示，GLI-1蛋白在鼻咽癌中存在明顯上調(diào)，上述結(jié)果表明Hh信號通路在鼻咽癌中存在導(dǎo)常激活。 通過特異性的信號通路抑制劑環(huán)巴胺（cyclopamine）對Hh鼻咽癌不同分化程度的細胞系CNE1、CNE2和HNE2的細胞株進行通路功能性干擾。CCK-8細胞增殖實驗顯示，CNE1、CNE2和HNE2在信號通路受抑制情況下，細胞生長和增殖受到一定程度的抑制。流式細胞儀檢測發(fā)現(xiàn)，cyclopamine抑制通路導(dǎo)致CNE1、CNE2和HNE2細胞凋亡增加。熒光定量PCR檢測通路干擾對目的基因的表達影響發(fā)現(xiàn)，下游基因PTCH1、Cyclin E表達下調(diào)。 本研究結(jié)果顯示，Hh信號通路在鼻咽癌中存在異常激活，通過對信號通路的干擾研究發(fā)現(xiàn)，Hh促進鼻咽癌細胞的增殖及凋亡，并對信號通路的靶標(biāo)基因表達具有抑制作用。本研究推動實現(xiàn)鼻咽癌從Hh信號通路作為診斷和分子靶標(biāo)的研究提供新的科學(xué)依據(jù)。
[Abstract]:Nasopharyngeal carcinoma (NPC) is a multifactorial inherited common head and neck tumor, its occurrence and development has the characteristics of multi-stage, concealment and so on, and it is one of the common malignant tumors in China. The high incidence area is mainly concentrated in five southern provinces (Guangdong, Guangxi, Hunan, Fujian, Jiangxi), known as Guangdong cancer. Although due to the progress of surgery and chemotherapy, due to the concealment of the location of the disease and the effectiveness of the detection index, there are still considerable difficulties in the diagnosis and treatment of the disease. Most patients found that the disease was diagnosed in the late stage of cancer. Therefore, there is an urgent need for a more reasonable and effective diagnosis and prognosis of nasopharyngeal carcinoma. Hedgehog (Hh) signaling pathway is found in an evolutionary conservative signaling pathway in Drosophila melanogaster. Its initial function is to regulate embryonic development, and then it is found that it is abnormally activated in a variety of human cancers and is involved in tumorigenesis, differentiation, invasion and metastasis. However, it is not clear whether there is abnormal expression of Hh signaling pathway in nasopharyngeal carcinoma and its functional effect in nasopharyngeal carcinoma. In this study, the expression of PTCH-1,SMO and GLI-1 genes in nasopharyngeal carcinoma (NPC) and nasopharyngitis (NPC) was detected by fluorescence quantitative PCR (FQ-PCR). The results showed that PTCH-1,SMO and GLI-1 genes were expressed in all nasopharyngeal tissues, while the results of fluorescence quantitative PCR showed that the expression of PTCH,GLI-1 was different in nasopharyngeal carcinoma tissues and nasopharyngeal inflammation tissues (P 0.05), PTCH expression was down-regulated. The expression of GLI-1 was up-regulated, but the expression of SMO gene was not different. Immunohistochemical analysis further showed that GLI-1 protein was up-regulated in nasopharyngeal carcinoma (NPC). These results indicated that Hh signaling pathway was often activated in nasopharyngeal carcinoma (NPC). Cyclopramine (cyclopamine), a specific signal pathway inhibitor, interfered with the pathway function of Hh nasopharyngeal carcinoma cell lines CNE1,CNE2 and HNE2 with different degrees of differentiation. CCK-8 cell proliferation test showed that CNE1, The growth and proliferation of CNE2 and HNE2 were inhibited to a certain extent under the condition of signal pathway inhibition. Flow cytometry showed that cyclopamine inhibitory pathway increased apoptosis of CNE1,CNE2 and HNE2 cells. Fluorescence quantitative PCR was used to detect the effect of pathway interference on the expression of target gene. It was found that the expression of downstream gene PTCH1,Cyclin E was down-regulated. The results showed that Hh signaling pathway was abnormally activated in nasopharyngeal carcinoma (NPC). Through the interference of signal pathway, it was found that Hh promoted the proliferation and apoptosis of nasopharyngeal carcinoma cells, and inhibited the expression of target gene in signal pathway. This study promotes the study of nasopharyngeal carcinoma from Hh signaling pathway as a diagnosis and molecular target to provide a new scientific basis.
【學(xué)位授予單位】：中山大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63

【參考文獻】

相關(guān)期刊論文 前5條

1 何志巍,謝鷺,許亮國,藍軻,劉衛(wèi)東,張玲,任彩萍,史建凌,周文,姚開泰;一個與人鼻咽癌相關(guān)新基因的克隆[J];科學(xué)通報;2000年11期

2 王紓宜;朱莉;李詩敏;林嵐;鄭斯馨;吳韻芳;;2005年WHO鼻咽部腫瘤的分類及鼻咽癌的TNM分期[J];中華耳鼻咽喉頭頸外科雜志;2007年08期

3 賴振南,黃應(yīng)桂,余忠華;nm23-H_1基因表達與鼻咽癌的早發(fā)轉(zhuǎn)移[J];腫瘤研究與臨床;2005年03期

4 李建萍 ,張小清 ,左克強 ,李植源;鼻咽癌p16基因表達與突變的對比研究及意義(英文)[J];中國現(xiàn)代醫(yī)學(xué)雜志;2005年10期

5 易俊林;高黎;黃曉東;李素艷;羅京偉;徐國鎮(zhèn);肖建平;蔡偉明;;鼻咽癌的臨床表現(xiàn)和預(yù)后[J];中國醫(yī)學(xué)科學(xué)院學(xué)報;2006年03期

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