骨橋蛋白在鼻咽癌組織中的表達(dá)及臨床意義
發(fā)布時(shí)間：2018-11-08 13:37
【摘要】： 背景：鼻咽癌(nasopharyngeal carcinoma, NPC)是一種原發(fā)于鼻咽部的惡性腫瘤,它的病因、發(fā)生、發(fā)展及治療等在諸方面均與其他頭頸腫瘤有極大地不同。NPC在中國(guó)的南方地區(qū)比其他地區(qū)更為常見(jiàn)。它的發(fā)病機(jī)制是一個(gè)復(fù)雜的過(guò)程,主要涉及到遺傳易感性、EB病毒感染和環(huán)境因素等,但是很多具體機(jī)制還不清楚。骨橋蛋白(osteopontin, OPN)是一種重要的多功能細(xì)胞外基質(zhì)磷酸化糖蛋白。近來(lái),研究表明OPN在多種腫瘤中都過(guò)度表達(dá),包括肺癌、乳腺癌、結(jié)腸直腸癌、胃癌、卵巢癌、黑色素瘤和間皮瘤等。OPN通過(guò)與廣泛分布在細(xì)胞表面上的受體相互作用,參與多種癌的許多生理和病理過(guò)程,主要涉及腫瘤的惡性轉(zhuǎn)化、浸潤(rùn)、轉(zhuǎn)移和發(fā)展等。甚至有人提出OPN可以作為腫瘤分子靶向治療的一個(gè)靶點(diǎn)。目前,OPN在NPC組織的表達(dá)情況及其意義尚有待于闡明。 目的：研究OPN在NPC組織標(biāo)本中表達(dá)情況并且探討其在NPC中的意義；為NPC的早期診斷和臨床治療提供參考指標(biāo)和理論基礎(chǔ)。 試驗(yàn)設(shè)計(jì)、材料和方法：用橫向調(diào)查的方法對(duì)NPC、慢性鼻咽炎組織和癌旁非癌組織進(jìn)行研究。用半定量逆轉(zhuǎn)錄多聚酶鏈?zhǔn)椒磻?yīng)(RT-PCR)的方法測(cè)定OPN mRNA在新鮮的冰凍組織中的相對(duì)表達(dá),mRNA的含量用圖像分析技術(shù)來(lái)定量；用免疫組織化學(xué)法(IHC)對(duì)石蠟包埋的組織切片中OPN的表達(dá)的程度、強(qiáng)度和定位進(jìn)行評(píng)估。統(tǒng)計(jì)分析OPN mRNA和OPN蛋白的表達(dá)和NPC患者臨床參數(shù)之間的關(guān)系。我們還用蛋白印跡法(WB)檢測(cè)了12對(duì)NPC癌組織和癌旁非癌組織的配對(duì)標(biāo)本中OPN蛋白的表達(dá)情況。 結(jié)果：從半定量RT-PCR的結(jié)果中我們得到OPN/GAPDH(mRNA灰度值)的均數(shù)(±標(biāo)準(zhǔn)差)在44例NPC組織中為0.80(±0.24),相比之下15例慢性炎癥組織中僅為0.32(±0.18),二組之間有統(tǒng)計(jì)學(xué)意義(P0.05)。IHC染色顯示OPN蛋白主要定位于腫瘤細(xì)胞的細(xì)胞漿中,其陽(yáng)性表達(dá)率在67例NPC組織中(88.1%%)顯著高于21例慢性鼻咽炎組織中的陽(yáng)性率(28.6%)(P0.05)。對(duì)以上兩個(gè)結(jié)果統(tǒng)計(jì)學(xué)分析顯示OPN mRNA和OPN蛋白的表達(dá)均和年齡和性別無(wú)關(guān)(P0.05),與腫瘤的T、N和臨床分期有關(guān)(P0.05)。WB結(jié)果顯示OPN蛋白的表達(dá)水平在癌組織中高于相對(duì)應(yīng)的癌旁非癌組織中的表達(dá),平均OPN/β-Actin(灰度值)分別為0.83(±0.16)和0.51(±0.21),二組有顯著差異(P0.05)。 結(jié)論：我們的研究結(jié)果表明OPN mRNA和OPN蛋白在NPC組織中過(guò)度表達(dá),推測(cè)它可能參與了NPC的惡性轉(zhuǎn)化；OPN mRNA和OPN蛋白的表達(dá)和NPC的臨床參數(shù)密切相關(guān),推測(cè)它在NPC的侵襲、轉(zhuǎn)移和進(jìn)展中起到某種作用。
[Abstract]:Background: nasopharyngeal carcinoma (nasopharyngeal carcinoma, NPC) is a primary malignant tumor in the nasopharynx. Development and treatment are significantly different from other head and neck tumors. NPC is more common in southern China than in other regions. Its pathogenesis is a complex process, mainly involving genetic susceptibility, EB virus infection and environmental factors, but many specific mechanisms are not clear. Osteopontin (osteopontin, OPN) is an important multifunctional extracellular matrix phosphorylated glycoprotein. Recently, studies have shown that OPN is overexpressed in a variety of tumors, including lung cancer, breast cancer, colorectal cancer, gastric cancer, ovarian cancer, melanoma and mesothelioma. OPN interacts with receptors widely distributed on the cell surface. Participate in many physiological and pathological processes of cancer, mainly involved in malignant transformation, invasion, metastasis and development. It has even been suggested that OPN can be used as a target for tumor molecular targeting therapy. At present, the expression of OPN in NPC and its significance need to be clarified. Objective: to study the expression of OPN in NPC tissues and its significance in NPC, and to provide a reference index and theoretical basis for the early diagnosis and clinical treatment of NPC. Design, materials and methods: NPC, chronic nasopharyngitis and adjacent non-cancerous tissues were studied by transverse survey. The relative expression of OPN mRNA in fresh frozen tissues was determined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and the content of mRNA was quantified by image analysis. The degree, intensity and localization of OPN expression in paraffin embedded tissue sections were evaluated by immunohistochemical (IHC). The relationship between the expression of OPN mRNA and OPN protein and the clinical parameters of NPC patients was analyzed statistically. We also detected the expression of OPN protein in 12 pairs of matched specimens of NPC and non-cancerous adjacent tissues by Western blot (WB). Results: from the results of semi-quantitative RT-PCR, we obtained that the mean value of OPN/GAPDH (鹵standard deviation) was 0.80 (鹵0.24) in 44 cases of NPC, compared with 0.32 (鹵0.18) in 15 cases of chronic inflammation. There was significant difference between the two groups (P0.05). IHC staining showed that OPN protein was mainly located in the cytoplasm of tumor cells. The positive expression rate in 67 cases of NPC (88.1%) was significantly higher than that of 21 cases of chronic nasopharyngitis (28.6%) (P0.05). Statistical analysis of the above two results showed that the expression of OPN mRNA and OPN protein was not related to age and sex (P0.05), but related to tumor T, N was correlated with clinical stage (P0.05). WB results showed that the expression of OPN protein in cancer tissues was higher than that in adjacent non-cancerous tissues. The average OPN/ 尾-Actin (gray value) was 0. 83 (鹵0. 16) and 0. 51 (鹵0. 21), respectively. There was significant difference between the two groups (P0.05). Conclusion: our results suggest that OPN mRNA and OPN proteins are overexpressed in NPC tissues and may be involved in the malignant transformation of NPC. The expression of OPN mRNA and OPN proteins is closely related to the clinical parameters of NPC. It is assumed that they play a role in the invasion, metastasis and progression of NPC.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R739.63
本文編號(hào)：2318654
[Abstract]:Background: nasopharyngeal carcinoma (nasopharyngeal carcinoma, NPC) is a primary malignant tumor in the nasopharynx. Development and treatment are significantly different from other head and neck tumors. NPC is more common in southern China than in other regions. Its pathogenesis is a complex process, mainly involving genetic susceptibility, EB virus infection and environmental factors, but many specific mechanisms are not clear. Osteopontin (osteopontin, OPN) is an important multifunctional extracellular matrix phosphorylated glycoprotein. Recently, studies have shown that OPN is overexpressed in a variety of tumors, including lung cancer, breast cancer, colorectal cancer, gastric cancer, ovarian cancer, melanoma and mesothelioma. OPN interacts with receptors widely distributed on the cell surface. Participate in many physiological and pathological processes of cancer, mainly involved in malignant transformation, invasion, metastasis and development. It has even been suggested that OPN can be used as a target for tumor molecular targeting therapy. At present, the expression of OPN in NPC and its significance need to be clarified. Objective: to study the expression of OPN in NPC tissues and its significance in NPC, and to provide a reference index and theoretical basis for the early diagnosis and clinical treatment of NPC. Design, materials and methods: NPC, chronic nasopharyngitis and adjacent non-cancerous tissues were studied by transverse survey. The relative expression of OPN mRNA in fresh frozen tissues was determined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), and the content of mRNA was quantified by image analysis. The degree, intensity and localization of OPN expression in paraffin embedded tissue sections were evaluated by immunohistochemical (IHC). The relationship between the expression of OPN mRNA and OPN protein and the clinical parameters of NPC patients was analyzed statistically. We also detected the expression of OPN protein in 12 pairs of matched specimens of NPC and non-cancerous adjacent tissues by Western blot (WB). Results: from the results of semi-quantitative RT-PCR, we obtained that the mean value of OPN/GAPDH (鹵standard deviation) was 0.80 (鹵0.24) in 44 cases of NPC, compared with 0.32 (鹵0.18) in 15 cases of chronic inflammation. There was significant difference between the two groups (P0.05). IHC staining showed that OPN protein was mainly located in the cytoplasm of tumor cells. The positive expression rate in 67 cases of NPC (88.1%) was significantly higher than that of 21 cases of chronic nasopharyngitis (28.6%) (P0.05). Statistical analysis of the above two results showed that the expression of OPN mRNA and OPN protein was not related to age and sex (P0.05), but related to tumor T, N was correlated with clinical stage (P0.05). WB results showed that the expression of OPN protein in cancer tissues was higher than that in adjacent non-cancerous tissues. The average OPN/ 尾-Actin (gray value) was 0. 83 (鹵0. 16) and 0. 51 (鹵0. 21), respectively. There was significant difference between the two groups (P0.05). Conclusion: our results suggest that OPN mRNA and OPN proteins are overexpressed in NPC tissues and may be involved in the malignant transformation of NPC. The expression of OPN mRNA and OPN proteins is closely related to the clinical parameters of NPC. It is assumed that they play a role in the invasion, metastasis and progression of NPC.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R739.63
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