磷脂酶Cγ-2参与的花生四烯酸的脂加氧酶-12代谢通路与喉鳞状细胞癌的关系及临床意义

发布时间：2018-01-08 17:02

本文关键词：磷脂酶Cγ-2参与的花生四烯酸的脂加氧酶-12代谢通路与喉鳞状细胞癌的关系及临床意义　出处：《河北医科大学》2013年硕士论文　论文类型：学位论文

【摘要】：目的：通过检测喉癌（laryngeal carcinoma）患者癌组织及其癌旁组织中磷脂酶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）的表达，分析其与肿瘤病理分级和临床分期的相互关系，探讨脂质代谢在喉鳞癌发生、发展过程所发挥的重要作用，从而为喉癌制定治疗方案提供分子生物学依据。方法：选择河北医科大学第二医院耳鼻喉科2011年2月至2012年5月收治的喉癌患者，共30例，均为男性，年龄53岁-70岁，平均60.15.3岁，术前未接受过放化疗，且无甲亢、甲状腺机能减退等疾病。所有患者病例资料完整，术前均经过病理活检证实为喉鳞状细胞癌。选取其癌组织和癌旁组织作为研究对象，其中癌旁组织取自距癌组织＞0.5cm处（经病理证实为正常组织）。按2002年国际抗癌联盟(UICC)标准进行临床分期，其中I期～II期18例，III期～Ⅳ期12例；按肿瘤分化程度将其分级，其中高分化鳞癌19例，中～低分化鳞癌11例，经病理确认有颈部淋巴结转移的14例，未转移的16例；按年龄将其分组，以60岁为标准，53-60岁组的标本19例，61-70岁组的标本11例。用免疫组化方法分析喉癌组织和癌旁组织中的PLCγ-2以及12-LOX的表达；用气相色谱联合质谱（gaschromatography/mass spectrometry, GC/MS）法对喉癌组织和癌旁组织中的AA含量进行测定。首先，每例手术标本取材后，一部分随即浸泡在10%的福尔马林液中进行固定，然后常规石蜡包埋，以5μm厚度连续切片，石蜡切片经常规二甲苯脱蜡，梯度酒精脱水，去除内源性过氧化物酶，微波抗原修复，以Streptavidin-perosidas法(SP法)分别行PLCγ-2和12-LOX免疫组织化学染色检测。继而DAB显色，苏木精复染，梯度酒精脱水、二甲苯透明、树胶封片。然后进行读片，数据分析。另一部分手术标本放入生理盐水中，送往实验室进行AA的提取和衍生化，用气相色谱联合质谱法进行分析和研究。所有实验数据均用SPSS l3.0统计软件进行分析，以P 0.05为差异有统计学意义。结果：PLCγ-2蛋白在喉癌组织中的阳性表达较癌旁组织中的表达增高，二者间差异有统计学意义（P 0.05）。12-LOX蛋白在喉癌组织中的阳性表达较癌旁组织中的表达增高，二者间差异有统计学意义（P 0.05）。AA在喉癌组织中的含量较癌旁组织中的含量偏高，二者间差异有统计学意义（P 0.05）。 PLCγ-2蛋白的阳性表达与喉鳞状细胞癌的临床分期及病理分级有着密切关系，I～II期喉癌组织中PLCγ-2的表达较III～Ⅳ期偏低，在高分化喉癌组织中PLCγ-2的表达较中～低分化喉癌组织中的表达偏低，两者间差异均具有统计学意义（P 0.05）。 12-LOX蛋白的阳性表达与喉鳞状细胞癌的临床分期和病理分级均有密切关系，I～II期喉癌组织中12-LOX的表达较III～Ⅳ期偏低，高分化喉癌组织中的表达较中～低分化喉癌组织中的表达偏低，二者间差异都具有统计学意义（P 0.05）。 AA含量与喉癌临床分期和病理分级均有密切关系，I～II期喉癌组织中AA的含量较III～Ⅳ期偏低，高分化喉癌组织中的表达较中～低分化喉癌组织中的表达偏低，二者间差异都具有统计学意义（P 0.05）。 PLCγ-2、12-LOX的蛋白阳性表达及AA的含量与癌细胞的淋巴结转移密切相关，淋巴结转移组中PLCγ-2、12-LOX的阳性表达明显高于淋巴结未转移组，淋巴结转移组中AA的含量高于淋巴结未转移组，且差异都有统计学意义（P 0.05）。 PLCγ-2和12-LOX的蛋白阳性表达与患者年龄无关（P＞0.05），AA含量与患者年龄无关（P＞0.05）。结论：PLCγ-2和12-LOX在喉癌组织中高表达，且与喉癌的临床分期、病理分级以及淋巴结转移关系密切，与年龄无关。肿瘤的临床侵犯范围越广，伴有淋巴结转移，，PLCγ-2和12-LOX的蛋白表达越高。而AA在癌组织中的含量较癌旁组织中的含量偏高，与喉癌的临床分期、病理分化程度、淋巴结转移相关，癌症临床分期越高、病理分化程度越差、伴有淋巴结转移，AA含量越高，而与年龄无关。上述结果提示，脂质代谢与喉癌的发生、发展和转移关系紧密，PLCγ-2、12-LOX和AA三者结合，可作为判断喉癌治疗效果和预后的一项指标，对临床肿瘤的诊断、治疗和预后评估提供相应的分子生物学依据。
[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.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R739.65

【参考文献】