本文選題：claudin-1 + cyclinB1�。� 參考：《山東大學(xué)》2015年博士論文

【摘要】：下咽癌在頭頸部惡性腫瘤中相對(duì)較少見,90%以上為鱗狀細(xì)胞癌；雖然發(fā)病率較低,但卻是惡性程度較高的腫瘤之一,其具體病因尚不明確,絕大多數(shù)病人有長(zhǎng)期大量的吸煙和飲酒史,考慮可能與長(zhǎng)期吸煙飲酒關(guān)系密切,臨床上主要表現(xiàn)為咽部不適、咽異物感、咽痛、吞咽疼痛、頸部包塊等,晚期可出現(xiàn)聲音嘶啞、吞咽困難、呼吸困難等,因早期癥狀不典型,同時(shí)下咽部有豐富的淋巴管網(wǎng),往往患者發(fā)現(xiàn)時(shí)已經(jīng)處于晚期,多伴有頸部淋巴結(jié)轉(zhuǎn)移和/或鄰近部位侵犯；目前多采用以手術(shù)治療為主,輔以放療、化療的綜合治療的方法,但患者預(yù)后仍然較差,國(guó)外報(bào)道5年的生存率只有30%。而早發(fā)現(xiàn)、早治療將有效的改善下咽癌患者的預(yù)后,因此尋找一些理想的腫瘤標(biāo)志物會(huì)幫助我們了解下咽癌腫瘤的生物學(xué)特性,從而有利于制定個(gè)性化的治療方案,改善腫瘤患者的預(yù)后。緊密連接是位于上皮細(xì)胞及內(nèi)皮細(xì)胞間的一種重要結(jié)構(gòu),在維持細(xì)胞穩(wěn)態(tài)、維系上皮細(xì)胞間黏附和保持細(xì)胞的極性中發(fā)揮重要作用,其主要封閉蛋白為claudin蛋白,其結(jié)構(gòu)及功能的變化直接受到claudin蛋白表達(dá)數(shù)量的變化所影響；目前研究認(rèn)為claudin蛋白可在多種惡性腫瘤中出現(xiàn)異常表達(dá),使細(xì)胞極性丟失,細(xì)胞間的黏附性下降,從而改變了緊密連接的正常結(jié)構(gòu)及功能,促使腫瘤細(xì)胞獲得侵襲力及轉(zhuǎn)移的潛能。claudin-1屬claudins蛋白家族成員之一,在多種腫瘤中出現(xiàn)異常表達(dá),研究表明,claudin-1在腎細(xì)胞癌、卵巢癌、胃腺癌和大腸癌中過度表達(dá),促進(jìn)了腫瘤發(fā)生發(fā)展的過程,而在對(duì)乳腺癌和肺腺癌的研究中,發(fā)現(xiàn)claudin-1表達(dá)顯著降低。這種差異性的表達(dá)方式已被證實(shí)與腫瘤的發(fā)生、發(fā)展密切相關(guān)。正常細(xì)胞的周期是在特定的細(xì)胞周期蛋白和細(xì)胞周期蛋白依賴性激酶的作用下,忠實(shí)而有序的啟動(dòng)和完成的,而不受控制的增殖是惡性腫瘤的標(biāo)志之一,細(xì)胞周期調(diào)節(jié)失控是細(xì)胞過度增殖及癌變的重要原因cyclinB1是一個(gè)經(jīng)典的細(xì)胞周期蛋白,通過改變細(xì)胞G2/M期的進(jìn)程,改變了細(xì)胞周期進(jìn)程,從而參與調(diào)控細(xì)胞周期檢查點(diǎn)。目前研究顯示,在多種惡性腫瘤細(xì)胞中,如在乳腺癌、胃癌、早期非小細(xì)胞肺癌、結(jié)腸直腸癌、前列腺癌中都發(fā)現(xiàn)cyclinBl(?)過表達(dá),cyclinB1失控性的表達(dá)與腫瘤細(xì)胞的轉(zhuǎn)化和惡性增殖密切相關(guān),而進(jìn)一步研究顯示,cyclinB1的過表達(dá)與食管癌、舌癌、肺癌患者的預(yù)后有關(guān),所以cyclinB1有可能成為一個(gè)潛在的腫瘤診斷標(biāo)志。惡性腫瘤的重要特性是浸潤(rùn)、擴(kuò)散和轉(zhuǎn)移,腫瘤的轉(zhuǎn)移是一個(gè)復(fù)雜的過程,一般可分為黏附、降解、移動(dòng)3個(gè)步驟,受到多種因素的影響,而通過血管及淋巴管轉(zhuǎn)移是多數(shù)腫瘤最常見、最主要的轉(zhuǎn)移方式,血管和淋巴管成為腫瘤轉(zhuǎn)移的重要通道,目前認(rèn)為在惡性腫瘤的過度生長(zhǎng)、局部侵襲及轉(zhuǎn)移過程中,新生的血管及淋巴管起到了極其重要的作用。大量的研究使腫瘤通過血管轉(zhuǎn)移得到了很大的發(fā)展；而淋巴管的轉(zhuǎn)移在腫瘤的轉(zhuǎn)移中也起著重要的作用,腫瘤細(xì)胞在原發(fā)灶快速生長(zhǎng),并從原發(fā)灶上脫離,浸潤(rùn)、進(jìn)入附近的微淋巴管,沿微淋巴管運(yùn)行至目標(biāo)淋巴結(jié),在目標(biāo)淋巴管內(nèi)進(jìn)行大量增殖,最終形成轉(zhuǎn)移淋巴結(jié)。隨著特異性的抗淋巴管內(nèi)皮細(xì)胞抗體的發(fā)現(xiàn),微淋巴管作為腫瘤侵襲、轉(zhuǎn)移的通道,在腫瘤的侵襲、轉(zhuǎn)移過程中的作用越來越被人們所重視。微淋巴管密度(microlymphatic vessel density, MLVD)成為評(píng)估惡性腫瘤轉(zhuǎn)移與預(yù)后的重要因子。綜上所述,我們發(fā)現(xiàn)claudin-1和cylinBl在下咽鱗狀細(xì)胞癌組織中的表達(dá)水平、與腫瘤臨床分期、病理學(xué)分級(jí)、患者臨床預(yù)后之間的相關(guān)性尚有待研究；claudin-1蛋白與下咽癌微淋巴管生成是否相關(guān)尚不清楚。本研究采用免疫組化和實(shí)時(shí)定量PCR的方法,測(cè)定claudin-1和cyclinBl在下咽癌組織及癌旁組織中的表達(dá)情況,分析claudin-1口cyclinBl與下咽鱗癌的臨床病理特征之間的關(guān)系,同時(shí)分析它們與患者預(yù)后之間的關(guān)系,采用免疫組化的方法檢測(cè)D2-40標(biāo)記的微淋巴管密度,研究claudin-1蛋白與下咽鱗癌淋巴管生成的關(guān)系,以期為下咽癌的早期診斷和預(yù)后判斷提供分子生物學(xué)指標(biāo),為尋求腫瘤靶向治療提供研究方向,同時(shí)應(yīng)用體外實(shí)驗(yàn)通過沉默claudin-1觀察對(duì)FaDu細(xì)胞增殖及侵襲的影響,進(jìn)一步研究claudin-1的生物學(xué)特性。第一部分claudin-1與cyclinBl在下咽癌中的表達(dá)及其臨床意義下咽鱗狀細(xì)胞癌的發(fā)病率雖然較低,但因原發(fā)腫瘤位置隱蔽,不易發(fā)現(xiàn),多數(shù)下咽癌患者在就診時(shí)已經(jīng)是晚期,往往已發(fā)生臨近部位侵犯和頸部淋巴結(jié)轉(zhuǎn)移,患者預(yù)后較差。claudin-1屬于claudins蛋白家族成員之一,在多種腫瘤中出現(xiàn)異常表達(dá),目前研究表明,這種異常表達(dá)與腫瘤的發(fā)生、發(fā)展有關(guān)；cyclinBl是一個(gè)經(jīng)典的細(xì)胞周期蛋白,目前發(fā)現(xiàn)cyclinBl失控性的表達(dá)與腫瘤細(xì)胞的轉(zhuǎn)化和惡性增殖密切相關(guān)。但是claudin-1與cyclinB1在下咽鱗癌中的表達(dá)還罕有報(bào)道。因此,我們首先研究claudin-1與cyclinB1在下咽鱗癌及癌旁組織中的表達(dá)情況,分析claudin-1和cyclinB1與下咽鱗癌的臨床病理特征之間的關(guān)系,同時(shí)分析它們與患者預(yù)后之間的關(guān)系。目的與方法：1、選取97例具有完整臨床病理資料的下咽鱗癌患者的組織學(xué)標(biāo)本為研究對(duì)象,并選取90例癌旁正常黏膜組織作為對(duì)照,采用免疫組化的方法從蛋白水平比較claudin-1和cyclinB1在下咽鱗癌及正常黏膜組織中的表達(dá)的差異,分析兩者在下咽鱗癌中的表達(dá)與臨床病理參數(shù)及患者預(yù)后之間的關(guān)系。2、選取30位下咽癌病人術(shù)后的癌組織及癌旁正常黏膜,采用RT-PCR方法,在mRNA水平檢測(cè)claudin-1口cyclinB1的表達(dá)并定量分析,探討兩者的相關(guān)性。結(jié)果：1.claudin-1與cyclinB1在下咽癌中均為高表達(dá)；claudin-1蛋白表達(dá)與腫瘤的分化程度和淋巴結(jié)轉(zhuǎn)移有關(guān)；cyclinB1蛋白表達(dá)與腫瘤的分化程度有關(guān)；而與其他臨床病理資料無明顯相關(guān)性。Kaplan-Meier分析顯示claudin-1與患者的生存率相關(guān)(P＝0.003)。claudin-1與cyclinB1呈明顯正相關(guān)關(guān)系。2.claudin-1 mRNA和cyclinB1mRNA在下咽癌組織中的表達(dá)量均高于癌旁黏膜組織中的表達(dá)量。Spearman等級(jí)分析結(jié)果顯示在腫瘤組織和癌旁黏膜組織中claudin-1mRNA與cyclinB1mRNA之間存在明顯正相關(guān)關(guān)系。結(jié)論：(1)claudin-1在下咽鱗狀細(xì)胞癌組織中呈高表達(dá),且與腫瘤的分化程度和淋巴結(jié)轉(zhuǎn)移有關(guān)；并與患者的生存率相關(guān),claudin-1低表達(dá)者有較高的生存率(2)cyclinB1在下咽鱗狀細(xì)胞癌組織中亦呈高表達(dá),并與腫瘤的分化程度有關(guān),與患者的生存率無關(guān)。(3)claudin-1和cyclinB1在下咽鱗狀細(xì)胞癌組織中的表達(dá)具有顯著正相關(guān)關(guān)系。第二部分claudin-1蛋白與下咽癌微淋巴管密度的關(guān)系研究下咽鱗癌發(fā)生部位比較隱匿,同時(shí)下咽部有豐富的淋巴管網(wǎng),容易較早出現(xiàn)頸部淋巴結(jié)轉(zhuǎn)移,臨床分期N1-N�；颊哳i部淋巴結(jié)轉(zhuǎn)移率為65%-80%,目前認(rèn)為,頸部淋巴結(jié)轉(zhuǎn)移成為下咽癌治療失敗的主要原因之一,嚴(yán)重影響了患者的預(yù)后。惡性腫瘤的重要特性包括浸潤(rùn)和轉(zhuǎn)移,而多數(shù)腫瘤通過淋巴管發(fā)生轉(zhuǎn)移,目前認(rèn)為淋巴管的生成在腫瘤的生長(zhǎng)、侵襲及轉(zhuǎn)移過程中發(fā)揮了極其重要的作用。微淋巴管密度(microlymphatic vessel density, MLVD)成為評(píng)估惡性腫瘤轉(zhuǎn)移與預(yù)后的重要因子。D2-40被認(rèn)為是目前發(fā)現(xiàn)的最具有特異性的淋巴管標(biāo)記物,廣泛的應(yīng)用于腫瘤微淋巴管生成的研究,我們前期研究中證實(shí)下咽鱗狀細(xì)胞癌中claudin-1蛋白表達(dá)情況與腫瘤淋巴結(jié)轉(zhuǎn)移有關(guān)。但claudin-1蛋白與下咽癌微淋巴管生成是否相關(guān)尚不清楚,是否通過微淋巴管生成導(dǎo)致了淋巴結(jié)的轉(zhuǎn)移尚待研究；本研究通過研究claudin-1蛋白與下咽鱗癌淋巴管生成的關(guān)系,探討下咽鱗癌淋巴轉(zhuǎn)移的機(jī)制。目的與方法：選取97例具有完整臨床病理資料的下咽鱗癌患者的組織學(xué)標(biāo)本為研究對(duì)象,并選取90例癌旁正常黏膜組織作為對(duì)照,采用免疫組化的方法檢測(cè)claudin-1 和微淋巴管密度在下咽鱗癌及正常黏膜組織中的表達(dá),分析微淋巴管密度與下咽鱗癌臨床病理資料的關(guān)系,分析claudin-1和微淋巴管密度在下咽鱗癌及正常黏膜組織中表達(dá)的相關(guān)性,從而探討下咽鱗癌淋巴轉(zhuǎn)移的機(jī)制。結(jié)果：1、下咽鱗癌中,癌巢周邊區(qū)的微淋巴管(MLVD)數(shù)目顯著高于癌巢中心區(qū)微淋巴管數(shù)目(MLVD)及癌旁正常粘膜組織中MLVD,兩者間差異均有統(tǒng)計(jì)學(xué)意義(P0.05)；而癌巢中心區(qū)MLVD與癌旁正常粘膜組織中MLVD相比,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。2、癌巢中心區(qū)的MLVD與患者各臨床指標(biāo)進(jìn)行相關(guān)分析均無統(tǒng)計(jì)學(xué)意義(P0.05)。癌巢周邊區(qū)的MLVD與組織病理學(xué)分級(jí)、臨床分期和淋巴結(jié)轉(zhuǎn)移等指標(biāo)顯著相關(guān),分析均有統(tǒng)計(jì)學(xué)意義(P0.05)。3、癌巢中心區(qū)的claudin-1表達(dá)陽性組的MLVD與陰性組相比差異無統(tǒng)計(jì)學(xué)意義,癌巢周邊區(qū)claudin-1表達(dá)陽性MLVD與陰性組MLVD相比有統(tǒng)計(jì)學(xué)意義(P0.05),相關(guān)分析表明在下咽鱗癌癌巢周邊區(qū)中claudin-1表達(dá)與MLVD之間呈正相關(guān)表達(dá)。結(jié)論：1、下咽癌癌巢周邊區(qū)微淋巴管密度和淋巴結(jié)轉(zhuǎn)移相關(guān)(P0.05),提示癌巢周邊區(qū)微淋巴管可能促進(jìn)了局部淋巴結(jié)的轉(zhuǎn)移。2、claudin-1陽性表達(dá)和陰性表達(dá)在癌巢周邊區(qū)的微淋巴管密度存在差異,提示高表達(dá)claudin-1可能會(huì)誘導(dǎo)腫瘤淋巴管生成,從而促進(jìn)淋巴結(jié)轉(zhuǎn)移。第三部分 沉默claudin-1在體外對(duì)細(xì)胞增殖及侵襲的影響我們?cè)谇捌谘芯恐邪l(fā)現(xiàn),claudin-1在下咽鱗狀細(xì)胞癌中呈現(xiàn)高表達(dá),與細(xì)胞分化及淋巴結(jié)轉(zhuǎn)移相關(guān),且claudin-1mRNA在下咽癌細(xì)胞中的含量增高,考慮claudin-1在下咽鱗癌中起了促進(jìn)腫瘤細(xì)胞增殖及局部侵襲轉(zhuǎn)移的功能,因此我們通過體外實(shí)驗(yàn),在FaDu細(xì)胞系中通過沉默claudin-1觀察FaDu細(xì)胞增殖及侵襲能力的變化,從而進(jìn)一步研究claudin-1的生物學(xué)特性。目的與方法：通過脂質(zhì)體2000轉(zhuǎn)染人咽癌細(xì)胞系(FaDu)沉默claudin-1,利用細(xì)胞培養(yǎng)技術(shù)進(jìn)行增殖實(shí)驗(yàn)和侵襲實(shí)驗(yàn)。結(jié)果：1、培養(yǎng)的FaDu細(xì)胞增殖活力良好。2、應(yīng)用RNA干擾技術(shù),沉默claudin-1 后, FaDu-siRNA瞬時(shí)沉默的效果良好,經(jīng)Western blot實(shí)驗(yàn)證實(shí)。3、沉默claudin-1后,FaDu-siRNA (沉默組)、FaDu-control(陰性對(duì)照組)以及FaDu未處理組,三組細(xì)胞均出現(xiàn)了不同程度的增殖現(xiàn)象,三組的吸光值在48小時(shí)后,沉默組與陰性對(duì)照組及未處理組比較,細(xì)胞的增殖明顯受影響,沉默組增殖能力降低,差異具有統(tǒng)計(jì)學(xué)意義(P0.01),而陰性對(duì)照組及未處理組之間比較,差異沒有統(tǒng)計(jì)學(xué)意義(P0.05)。4、侵襲實(shí)驗(yàn)表明：進(jìn)入小室的細(xì)胞數(shù),沉默組與陰性對(duì)照組及空白對(duì)照組分別為15士6.45、47士6.36、50士8.73,差異有統(tǒng)計(jì)學(xué)意義(P0.001)；沉默claudin-1后,細(xì)胞的侵襲能力明顯下降,FaDu-control以及FaDu未處理組比FaDu-siRNA組具有更強(qiáng)的侵襲能力。結(jié)論：體外實(shí)驗(yàn)顯示claudin-1能促進(jìn)下咽癌細(xì)胞的增殖、黏附及侵襲。
[Abstract]:Hypopharyngeal carcinoma is relatively rare in the malignant tumor of the head and neck, and more than 90% is squamous cell carcinoma. Although the incidence is low, it is one of the most malignant tumors. The specific cause is not clear. Most of the patients have a long history of smoking and drinking, which may be closely related to long term smoking and drinking. For pharynx discomfort, pharynx sensation, pharynx pain, swallowing pain, neck lump and so on, at the late stage can appear hoarseness, dysphagia, dyspnea and so on. The early symptoms are not typical, and the lower pharynx has a rich lymphatic network. With the method of surgical treatment, combined with radiotherapy and chemotherapy, the prognosis of patients is still poor. The survival rate of 5 years in foreign countries is only 30%.. Early treatment will effectively improve the prognosis of hypopharynx cancer patients, so finding some ideal tumor markers will help us understand the biological characteristics of hypopharyngeal cancer. It is beneficial to develop a personalized treatment scheme and improve the prognosis of cancer patients. Close connection is an important structure between epithelial cells and endothelial cells. It plays an important role in maintaining cell homeostasis, maintaining the adhesion of epithelial cells and maintaining the polarity of cells. The main closed protein is claudin protein, its structure and function. The changes in the expression of claudin protein are directly affected by the changes in the number of expression of the protein. The present study suggests that the abnormal expression of claudin protein in a variety of malignant tumors can cause the loss of cell polarity and the decrease of intercellular adhesion, thus changing the normal structure and function of tightly connected cells, and promoting the potential of invasion and metastasis of tumor cells,.C Laudin-1 is one of the members of the claudins protein family. Abnormal expression in a variety of tumors shows that claudin-1 is overexpressed in renal cell carcinoma, ovarian cancer, gastric adenocarcinoma and colorectal cancer, which promotes the process of tumor development. In the study of breast and lung adenocarcinoma, it is found that the expression of claudin-1 is significantly reduced. The expression pattern has been proved to be closely related to the occurrence and development of tumors. The cycle of normal cells is initiated and completed faithfully and orderly under the action of specific cyclin and cyclin dependent kinase, and uncontrolled proliferation is one of the markers of malignant tumor, and the control of cell cycle control is the overgrowth of cells. The important cause of colonization and carcinogenesis cyclinB1 is a classic cycyclin protein that changes the process of cell cycle by changing the process of G2/M phase and participates in the regulation of cell cycle checkpoints. Current studies show that in many malignant tumor cells, such as breast, gastric, early non small cell lung cancer, colorectal cancer, prostate CyclinBl (?) overexpression is found in cancer, and the expression of cyclinB1 out of control is closely related to the transformation and proliferation of tumor cells. Further studies have shown that the overexpression of cyclinB1 is associated with the prognosis of esophageal, tongue cancer, and lung cancer patients, so cyclinB1 may be a potential diagnostic marker. The important characteristics of malignant tumor It is infiltration, diffusion and metastasis, the metastasis of tumor is a complicated process, which can be divided into 3 steps, adhesion, degradation and movement, which are affected by many factors, and the most common and most important way of metastasis is the most common mode of metastasis through blood vessels and lymphatic metastasis, and the blood vessels and drenching tube are the important channels of tumor metastasis. The neovascularization and lymphatic vessels play an important role in the overgrowth of the tumor, local invasion and metastasis. A large number of studies have made the tumor develop greatly through the metastasis of blood vessels, and the metastasis of the lymphatic tube plays an important role in the metastasis of the tumor. The tumor cells grow rapidly in the primary focus and are from the primary focus. Disconnect, infiltrate, enter the nearby microlymphatics, run along the microlymphatics to the target lymph nodes, proliferate in the target lymphatics, and eventually form a metastatic lymph node. With the specific anti lymphatic endothelial cell antibody, the microlymphatic tube acts as a tumor invasion, the transfer channel, the role of the tumor in the invasion and metastasis process. Microlymphatic vessel density (MLVD) is an important factor in the assessment of metastasis and prognosis of malignant tumors. To sum up, we have found that the expression level of claudin-1 and cylinBl in the hypopharyngeal squamous cell carcinoma tissue is associated with the tumor stage, pathological grade, and the clinical prognosis of the tumor. The correlation between claudin-1 protein and the formation of microlymphatic vessels in hypopharyngeal carcinoma is not clear. The present study uses immunohistochemical and real-time quantitative PCR methods to determine the expression of claudin-1 and cyclinBl in hypopharyngeal and paracancerous tissues, and to analyze the clinicopathological features of claudin-1 mouth cyclinBl and hypopharyngeal squamous cell carcinoma. The relationship between them and the prognosis of patients was analyzed. The microlymphatic density of D2-40 markers was detected by immunohistochemical method, and the relationship between claudin-1 protein and the formation of lymphatic vessels in hypopharyngeal squamous cell carcinoma was studied in order to provide molecular biological indicators for the early diagnosis and prognosis of hypopharyngeal carcinoma and provide a study for the search for tumor targeting therapy. At the same time, the effects of claudin-1 observation on proliferation and invasion of FaDu cells were also applied in vitro, and the biological characteristics of claudin-1 were further studied. The first part of the expression of claudin-1 and cyclinBl in hypopharyngeal carcinoma and the incidence of squamous cell carcinoma of the pharynx in the clinical significance were low, but the location of the primary tumor was not easy to occur because of the location of the primary tumor. Now, most of the patients with hypopharyngeal cancer are in the advanced stage, and often have the invasion of adjacent areas and cervical lymph node metastasis. The poor prognosis of the patients is one of the members of the claudins protein family, which is abnormal expression in a variety of tumors. The present study shows that this abnormal expression is related to the development of the tumor and the development of the tumor; cyclinBl is the.Claudin-1 A classic cycyclin protein has been found to be closely related to the expression of cyclinBl out of control with the transformation and proliferation of tumor cells. However, the expression of claudin-1 and cyclinB1 in hypopharyngeal squamous cell carcinoma is rare. Therefore, we first study the expression of claudin-1 and cyclinB1 in hypopharyngeal squamous carcinoma and para cancer tissues, and analyze Claudi The relationship between n-1 and cyclinB1 with the clinicopathological features of hypopharyngeal squamous cell carcinoma and the relationship between them and the prognosis of the patients. Objective and methods: 1, 97 cases of hypopharyngeal squamous cell carcinoma with complete clinicopathological data were selected as the subjects, and 90 cases of normal mucosa adjacent to the cancer were selected as the control, and the immunological group was used. The expression of claudin-1 and cyclinB1 in hypopharyngeal squamous cell carcinoma and normal mucosal tissue was compared from the protein level. The relationship between the expression of the two in the hypopharyngeal squamous cell carcinoma and the clinicopathological parameters and the prognosis of the patients was analyzed.2. The cancer tissues and the normal mucosa adjacent to the cancer in 30 patients with hypopharynx cancer were selected and the RT-PCR method was used in mRNA water. The expression and quantitative analysis of the claudin-1 mouth cyclinB1 were measured and analyzed. Results: both 1.claudin-1 and cyclinB1 were highly expressed in the hypopharyngeal carcinoma; the expression of claudin-1 protein was related to the degree of differentiation and lymph node metastasis; the expression of cyclinB1 protein was related to the degree of differentiation of the tumor; and it was associated with other clinicopathological data. No significant correlation.Kaplan-Meier analysis showed that claudin-1 was associated with the survival rate of patients (P = 0.003).Claudin-1 and cyclinB1 had a significant positive correlation between.2.claudin-1 mRNA and cyclinB1mRNA in hypopharyngeal carcinoma tissues, which were higher than those in the paracancerous mucosa. There is a significant positive correlation between claudin-1mRNA and cyclinB1mRNA in mucous tissue. Conclusion: (1) claudin-1 is highly expressed in the hypopharyngeal squamous cell carcinoma and is related to the degree of differentiation and lymph node metastasis, and is associated with the survival rate of the patients. The low expression of claudin-1 has a higher survival rate (2) cyclinB1 in the hypopharyngeal squamous cell. The expression of claudin-1 and cyclinB1 in the tissues of hypopharyngeal squamous cell carcinoma has a significant positive correlation. (3) the relationship between the expression of the second part of the hypopharyngeal squamous cell carcinoma and the relationship between the density of the microlymphatic tube of the hypopharynx cancer and the location of the hypopharyngeal squamous cell carcinoma is relatively concealed. At the same time, there is a rich lymphatic network in the lower pharynx, which is prone to cervical lymph node metastasis earlier. The cervical lymph node metastasis rate of N1-N. patients is 65%-80%. It is considered that cervical lymph node metastasis is one of the main reasons for the failure of hypopharyngeal cancer treatment, which seriously affects the prognosis of the patients. Microlymphatic vessel density (MLVD) has become the most important factor in the assessment of metastasis and prognosis of malignant tumors..D2-40 is considered to be the most important factor to be found at present. Specific lymphatic markers are widely used in the study of the formation of tumor microlymphatics. Our previous study has confirmed that the expression of claudin-1 protein in hypopharyngeal squamous cell carcinoma is associated with tumor lymph node metastasis. However, whether claudin-1 protein is associated with the formation of the microlymphatics of hypopharyngeal carcinoma is not clear, and whether the microlymphatic vessels are generated by microlymphatic formation In this study, the mechanism of lymphatic metastasis of hypopharyngeal squamous cell carcinoma was investigated by studying the relationship between claudin-1 protein and the formation of lymphatic vessels in hypopharyngeal squamous cell carcinoma. Objective and methods: to select 97 cases of hypopharyngeal squamous cell carcinoma with complete clinicopathological data and 90 cases of normal paracancerous normal. As a control, the expression of claudin-1 and microlymphatic density in hypopharyngeal squamous cell carcinoma and normal mucous tissue were detected by immunohistochemical method. The relationship between the density of microlymphatic tube and the clinicopathological data of hypopharyngeal squamous cell carcinoma was analyzed. The correlation between the expression of claudin-1 and the density of microlymphatic tube in the hypopharyngeal squamous cell carcinoma and normal mucosa was analyzed. The mechanism of lymphatic metastasis of hypopharyngeal squamous cell carcinoma was investigated. Results: 1, the number of microlymphatic tubes (MLVD) in the surrounding area of the nests of the hypopharyngeal squamous cell carcinoma is significantly higher than the number of microlymphatics (MLVD) in the center of the carcinoma nest and the MLVD in the normal mucosa adjacent to the carcinoma (P0.05), while MLVD in the center of the carcinoma nest and the normal mucosa adjacent to the carcinoma are in the mucosa. Compared with MLVD, there was no significant difference (P0.05).2. There was no statistical significance in the correlation analysis between the MLVD of the cancer nest center and the clinical indexes of the patients (P0.05). The MLVD in the surrounding area of the nests was significantly correlated with the histopathological classification, the clinical stage and lymph node metastasis, and the analysis was statistically significant (P0.05).3, and claudin in the center of the cancer nest. There was no significant difference in the MLVD in the -1 positive group compared with the negative group. The claudin-1 expression positive MLVD in the surrounding area of the nests was statistically significant compared with the negative group MLVD (P0.05). The correlation analysis showed that the expression of Claudin-1 in the nests surrounding the hypopharyngeal squamous cell carcinoma was positively correlated with MLVD. Conclusion: 1, the microlymphatic lymph nodes around the nests of the hypopharyngeal carcinoma are microlymphatic. The correlation between tube density and lymph node metastasis (P0.05) suggests that the microlymphatics in the surrounding area of the carcinoma nest may promote the metastasis of local lymph nodes. The density of claudin-1 positive and negative expression in the peripheral region of the carcinoma nest is different, suggesting that high expression of claudin-1 may induce lymphatic angiogenesis and promote lymph node metastasis. Third The effect of partial silencing of claudin-1 on cell proliferation and invasion in vitro, we found that claudin-1 is highly expressed in hypopharyngeal squamous cell carcinoma, related to cell differentiation and lymph node metastasis, and the content of claudin-1mRNA in hypopharyngeal cancer cells is higher, considering that claudin-1 promotes tumor cells in hypopharyngeal squamous cell carcinoma. The function of proliferation and local invasion and metastasis, therefore, we observed the proliferation of FaDu cells in FaDu cell line by silencing Claudin-1 in vitro.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R739.63

【共引文獻(xiàn)】

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