本文關(guān)鍵詞：磷脂酶Cγ-2參與的花生四烯酸的脂加氧酶-12代謝通路與喉鱗狀細(xì)胞癌的關(guān)系及臨床意義　出處：《河北醫(yī)科大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 喉癌laryngeal carcinoma 磷脂酶Cγ-2phospholipase Cγ-2 PLCγ-2 血小板型12-LOX lipoxygenase-12 花生四烯酸arachidonic acid AA

【摘要】：目的：通過檢測喉癌（laryngeal carcinoma）患者癌組織及其癌旁組織中磷脂酶Cγ-2（phospholipase Cγ-2, PLCγ-2）、血小板型12-LOX（lipoxygenase-12,12-LOX）以及花生四烯酸（arachidonic acid, AA）的表達(dá)，分析其與腫瘤病理分級和臨床分期的相互關(guān)系，探討脂質(zhì)代謝在喉鱗癌發(fā)生、發(fā)展過程所發(fā)揮的重要作用，從而為喉癌制定治療方案提供分子生物學(xué)依據(jù)。 方法：選擇河北醫(yī)科大學(xué)第二醫(yī)院耳鼻喉科2011年2月至2012年5月收治的喉癌患者，共30例，均為男性，年齡53歲-70歲，平均60.15.3歲，術(shù)前未接受過放化療，且無甲亢、甲狀腺機(jī)能減退等疾病。所有患者病例資料完整，術(shù)前均經(jīng)過病理活檢證實為喉鱗狀細(xì)胞癌。選取其癌組織和癌旁組織作為研究對象，其中癌旁組織取自距癌組織＞0.5cm處（經(jīng)病理證實為正常組織）。按2002年國際抗癌聯(lián)盟(UICC)標(biāo)準(zhǔn)進(jìn)行臨床分期，其中I期～I(xiàn)I期18例，III期～Ⅳ期12例；按腫瘤分化程度將其分級，其中高分化鱗癌19例，中～低分化鱗癌11例，經(jīng)病理確認(rèn)有頸部淋巴結(jié)轉(zhuǎn)移的14例，未轉(zhuǎn)移的16例；按年齡將其分組，以60歲為標(biāo)準(zhǔn)，53-60歲組的標(biāo)本19例，61-70歲組的標(biāo)本11例。用免疫組化方法分析喉癌組織和癌旁組織中的PLCγ-2以及12-LOX的表達(dá)；用氣相色譜聯(lián)合質(zhì)譜（gaschromatography/mass spectrometry, GC/MS）法對喉癌組織和癌旁組織中的AA含量進(jìn)行測定。首先，每例手術(shù)標(biāo)本取材后，一部分隨即浸泡在10%的福爾馬林液中進(jìn)行固定，然后常規(guī)石蠟包埋，以5μm厚度連續(xù)切片，石蠟切片經(jīng)常規(guī)二甲苯脫蠟，梯度酒精脫水，去除內(nèi)源性過氧化物酶，微波抗原修復(fù)，以Streptavidin-perosidas法(SP法)分別行PLCγ-2和12-LOX免疫組織化學(xué)染色檢測。繼而DAB顯色，蘇木精復(fù)染，梯度酒精脫水、二甲苯透明、樹膠封片。然后進(jìn)行讀片，數(shù)據(jù)分析。另一部分手術(shù)標(biāo)本放入生理鹽水中，送往實驗室進(jìn)行AA的提取和衍生化，用氣相色譜聯(lián)合質(zhì)譜法進(jìn)行分析和研究。所有實驗數(shù)據(jù)均用SPSS l3.0統(tǒng)計軟件進(jìn)行分析，以P 0.05為差異有統(tǒng)計學(xué)意義。 結(jié)果：PLCγ-2蛋白在喉癌組織中的陽性表達(dá)較癌旁組織中的表達(dá)增高，二者間差異有統(tǒng)計學(xué)意義（P 0.05）。12-LOX蛋白在喉癌組織中的陽性表達(dá)較癌旁組織中的表達(dá)增高，二者間差異有統(tǒng)計學(xué)意義（P 0.05）。AA在喉癌組織中的含量較癌旁組織中的含量偏高，二者間差異有統(tǒng)計學(xué)意義（P 0.05）。 PLCγ-2蛋白的陽性表達(dá)與喉鱗狀細(xì)胞癌的臨床分期及病理分級有著密切關(guān)系，I～I(xiàn)I期喉癌組織中PLCγ-2的表達(dá)較III～Ⅳ期偏低，在高分化喉癌組織中PLCγ-2的表達(dá)較中～低分化喉癌組織中的表達(dá)偏低，兩者間差異均具有統(tǒng)計學(xué)意義（P 0.05）。 12-LOX蛋白的陽性表達(dá)與喉鱗狀細(xì)胞癌的臨床分期和病理分級均有密切關(guān)系，I～I(xiàn)I期喉癌組織中12-LOX的表達(dá)較III～Ⅳ期偏低，高分化喉癌組織中的表達(dá)較中～低分化喉癌組織中的表達(dá)偏低，二者間差異都具有統(tǒng)計學(xué)意義（P 0.05）。 AA含量與喉癌臨床分期和病理分級均有密切關(guān)系，I～I(xiàn)I期喉癌組織中AA的含量較III～Ⅳ期偏低，高分化喉癌組織中的表達(dá)較中～低分化喉癌組織中的表達(dá)偏低，二者間差異都具有統(tǒng)計學(xué)意義（P 0.05）。 PLCγ-2、12-LOX的蛋白陽性表達(dá)及AA的含量與癌細(xì)胞的淋巴結(jié)轉(zhuǎn)移密切相關(guān)，淋巴結(jié)轉(zhuǎn)移組中PLCγ-2、12-LOX的陽性表達(dá)明顯高于淋巴結(jié)未轉(zhuǎn)移組，淋巴結(jié)轉(zhuǎn)移組中AA的含量高于淋巴結(jié)未轉(zhuǎn)移組，且差異都有統(tǒng)計學(xué)意義（P 0.05）。 PLCγ-2和12-LOX的蛋白陽性表達(dá)與患者年齡無關(guān)（P＞0.05），AA含量與患者年齡無關(guān)（P＞0.05）。 結(jié)論：PLCγ-2和12-LOX在喉癌組織中高表達(dá)，且與喉癌的臨床分期、病理分級以及淋巴結(jié)轉(zhuǎn)移關(guān)系密切，與年齡無關(guān)。腫瘤的臨床侵犯范圍越廣，伴有淋巴結(jié)轉(zhuǎn)移，，PLCγ-2和12-LOX的蛋白表達(dá)越高。而AA在癌組織中的含量較癌旁組織中的含量偏高，與喉癌的臨床分期、病理分化程度、淋巴結(jié)轉(zhuǎn)移相關(guān)，癌癥臨床分期越高、病理分化程度越差、伴有淋巴結(jié)轉(zhuǎn)移，AA含量越高，而與年齡無關(guān)。上述結(jié)果提示，脂質(zhì)代謝與喉癌的發(fā)生、發(fā)展和轉(zhuǎn)移關(guān)系緊密，PLCγ-2、12-LOX和AA三者結(jié)合，可作為判斷喉癌治療效果和預(yù)后的一項指標(biāo)，對臨床腫瘤的診斷、治療和預(yù)后評估提供相應(yīng)的分子生物學(xué)依據(jù)。
[Abstract]:Objective: through the detection of laryngeal cancer (laryngeal carcinoma) phospholipase C cancer tissue and its adjacent tissues in gamma -2 (phospholipase C -2 PLC gamma, gamma -2), platelet 12-LOX (lipoxygenase-12,12-LOX) and four arachidonic acid (arachidonic acid, AA) expression, to analyze the relationship between tumor staging and pathological grading and clinical. Discussion on lipid metabolism in laryngeal squamous cell carcinoma and play an important role in the development process, so as to laryngeal cancer treatment plan provide molecular basis.
Methods: the second hospital of Hebei Medical University from February 2011 to May 2012 admitted to Department of ENT patients, 30 cases were male, aged 53 -70 years old, the average age of 60.15.3, had not received preoperative chemotherapy, and no hyperthyroidism, hypothyroidism and other diseases. All patients with complete clinical data, preoperative biopsy after laryngeal squamous cell carcinoma. The carcinoma tissues and adjacent tissues as the research object, the adjacent tissue from the cancerous tissue over 0.5cm (pathologically confirmed normal tissue). According to the 2002 UICC (UICC) standard clinical stage, the I stage to II stage 18 cases, stage III ~ IV 12 cases according to the degree of tumor differentiation; the classification, including 19 cases of high differentiated squamous cell carcinoma, to poorly differentiated squamous cell carcinoma in 11 cases, 14 cases of pathologically confirmed cervical lymph node metastasis, 16 cases without metastasis; by age grouping, with 60 years as the standard Standard, 19 cases of 53-60 years old group of specimens, the age group of 61-70 specimens in 11 cases. PLC gamma analysis of laryngeal carcinoma and adjacent tissues -2 and 12-LOX expression by immunohistochemical method; gas chromatography combined with mass spectrometry (gaschromatography/mass spectrometry GC/MS) to determine the content of AA in laryngeal carcinoma and adjacent tissues the method. First, each operation specimens were immediately, a part of immersion were fixed in 10% formalin, and then embedded in paraffin, with 5 mu m thick serial sections of paraffin sections of regular xylene dewaxing, gradient alcohol dehydration, removal of endogenous peroxidase, microwave antigen repair, by the method of Streptavidin-perosidas (SP method) were performed PLC gamma -2 and 12-LOX immunohistochemical staining. Then DAB staining, hematoxylin staining, gradient alcohol dehydration, xylene transparent balata. Then read the piece, another part of the data analysis. The specimens were put into normal saline, sent to the laboratory for AA extraction and derivatization, and analyzed by gas chromatography and mass spectrometry. All the experimental data were analyzed by SPSS l3.0 software. The difference between P 0.05 was statistically significant.
Results: the increased positive expression of PLC gamma -2 protein in laryngeal cancer tissues than in cancer tissues, the difference was statistically significant between the two (P 0.05) increased the expression of.12-LOX protein in laryngeal carcinoma tissues than in cancer tissues, the difference was statistically significant between the two (P 0.05) high content the content of.AA in laryngeal carcinoma tissues than in adjacent tissues, the difference was statistically significant between the two (P 0.05).
Clinical expression and laryngeal squamous cell carcinoma PLC protein gamma -2 stages and pathological grades are closely related, I ~ II in laryngeal carcinoma in the expression of PLC gamma -2 than III ~ IV is low, low expression of poorly differentiated laryngeal carcinoma in high differentiated laryngeal carcinoma PLC expression of -2 than in ~, the difference was statistically significant (P 0.05).
The staging and pathological grading were closely related to clinical expression and laryngeal squamous cell carcinoma 12-LOX protein, the expression of I ~ 12-LOX in laryngeal carcinoma stage II ~ IV is lower than III, low expression in poorly differentiated laryngeal cancer tissues differentiated laryngeal carcinoma than in ~, are statistically significant difference between the two (P 0.05).
The staging and pathological grading were closely related to the content of AA and I in laryngeal carcinoma, laryngeal carcinoma in stage II ~ AA ~ IV is lower than III, low expression in poorly differentiated laryngeal cancer tissues differentiated laryngeal carcinoma than in ~, are statistically significant differences between the two (P 0.05).
The content of cancer cells and the expression of AA and PLC gamma of -2,12-LOX is closely related to lymph node metastasis, lymph node metastasis positive group PLC gamma -2,12-LOX was significantly higher than that in the group without lymph node metastasis and the group without lymph node metastasis group was AA higher than that of lymph nodes, and the difference was statistically significant (all P 0.05).
The protein positive expression of PLC gamma -2 and 12-LOX was not related to the age of the patients (P > 0.05), and the content of AA was not related to the age of the patients (P > 0.05).
Conclusion: PLC gamma -2 and high expression of 12-LOX in laryngeal carcinoma, and the clinical and pathological grading and staging of laryngeal carcinoma, lymph node metastasis, regardless of age. The clinical tumor invasion range more widely, lymph node metastasis, PLC gamma -2 and 12-LOX protein expression is higher in cancer tissues and the content. The AA is in cancer tissue was higher, and the clinical staging of laryngeal carcinoma, tumor differentiation, lymph node metastasis and clinical staging of cancer is higher, the degree of pathological differentiation worse, lymph node metastasis, the higher the content of AA, and has nothing to do with age. These results suggest that lipid metabolism and the occurrence of laryngeal carcinoma. The development and transfer of the close relationship between PLC gamma -2,12-LOX and AA combination of the three, can be used as an index to the effect and prognosis of laryngeal carcinoma, the diagnosis of clinical tumor and provide molecular basis for the treatment and prognosis evaluation.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R739.65

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