發(fā)布時間：2018-07-23 19:25

【摘要】：目的:本項研究主要利用免疫組織化學(xué)(Immunohistochemistry)和蛋白質(zhì)印跡(Western Blot)兩種實驗方法來檢測內(nèi)源性硫化氫合成酶胱硫醚-γ-裂解酶(CSE)在行肺癌根治性切除術(shù)后并經(jīng)過病理驗證的肺癌組織與癌旁正常組織(此癌旁正常組織距離癌組織至少5cm以上)兩組病例標(biāo)本中的表達(dá)水平及對其臨床意義進(jìn)行研究。方法:本項實驗研究按照選取標(biāo)準(zhǔn)嚴(yán)格選取了從2010年1月?2012年1月期間在青島大學(xué)附屬青島市市立醫(yī)院醫(yī)院行肺癌根治手術(shù)后的肺癌病理標(biāo)本125例和距離癌組織5 cm以上癌旁正常肺組織標(biāo)本125例(其中病理組織標(biāo)本的結(jié)果已經(jīng)由青島大學(xué)附屬青島市市立醫(yī)院醫(yī)院病理科閱片后明確診斷)。所有選取病例手術(shù)前均未實行化學(xué)藥物和放射治療。應(yīng)用免疫組化技術(shù)和Western Blot檢測兩種實驗技術(shù)檢測胱硫醚-γ-裂解酶在125例肺癌組織及相應(yīng)癌旁正常黏膜的表達(dá)情況,并通過門診回訪或者電話咨詢進(jìn)行隨訪。利用SPSS 13.0統(tǒng)計學(xué)軟件分析本實驗研究相關(guān)資料:肺癌組織胱硫醚-γ-裂解酶的表達(dá)和其在癌旁正常肺癌組織中的表達(dá)存在明顯差異,胱硫醚-γ-裂解酶的表達(dá)情況與肺癌患者病理及臨床因素之間的關(guān)系。采用Kaplan-Meier法進(jìn)行生存分析并繪制生存曲線圖,用Log-rank檢驗其生存差異性。其中P0.05時表示差異存在統(tǒng)計學(xué)意義。結(jié)果:(1)胱硫醚-γ-裂解酶在肺癌組織明顯表達(dá),陽性表達(dá)主要在表達(dá)細(xì)胞質(zhì)中,呈棕黃色。胱硫醚-γ-裂解酶在肺癌組織和癌旁正常肺癌組織中的表達(dá)水平存在差異:肺癌組織中胱硫醚-γ-裂解酶的表達(dá)水平顯著高于其在癌旁正常組織的表達(dá),兩者之間表達(dá)水平差異有統(tǒng)計學(xué)意義(P0.05)。同時蛋白質(zhì)印跡Western Blot法檢測再次證明肺癌組織中胱硫醚-γ-裂解酶的表達(dá)水平明顯比肺癌旁正常組織中的表達(dá)水平高。(2)在肺癌組織中胱硫醚-γ-裂解酶的表達(dá)水平與病理及臨床因素之間的關(guān)系和肺癌患者5年生存率均存在差異:胱硫醚-γ-裂解酶的表達(dá)情況與TNM分期情況、周圍淋巴結(jié)發(fā)生轉(zhuǎn)移與否、腫瘤大小、腫瘤分化程度有關(guān)(P0.05);胱硫醚-γ-裂解酶低表達(dá)的肺癌患者5年生存率明顯高于胱硫醚-γ-裂解酶高表達(dá)的肺癌患者(P0.05)。結(jié)論:胱硫醚-γ-裂解酶在肺癌組織中呈現(xiàn)高水平表達(dá),可能在肺癌的發(fā)生、浸潤轉(zhuǎn)移等演變過程中發(fā)揮重要作用,胱硫醚-γ-裂解酶在肺癌細(xì)胞中的表達(dá)及胱硫醚-γ-裂解酶對肺癌細(xì)胞增殖和凋亡的影響將有助于了解胱硫醚-γ-裂解酶對肺癌腫瘤中的生物學(xué)效應(yīng),可為以后胱硫醚-γ-裂解酶和肺癌方面的更深層次的研究提供一定程度的研究思路和理論基礎(chǔ),同時對分子致病機(jī)制的深入研究可為肺癌的預(yù)防與治療提供一些策略和手段,因此以胱硫醚-γ-裂解酶為靶點的研究新思路將可作為評估肺癌早期發(fā)生及預(yù)后情況的參考指標(biāo)。
[Abstract]:Objective: to detect endogenous hydrogen sulfide synthase cystathithione- 緯 -lyase (CSE) by immunohistochemical (Immunohistochemistry) and Western blot (Western Blot) after radical resection of lung cancer. The expression level and clinical significance of the two groups of lung cancer tissues and adjacent normal tissues (at least above 5cm) were studied. Methods: according to the selection criteria, 125 lung cancer specimens from January 2010 to January 2012 after radical lung cancer surgery were selected from Qingdao City Hospital affiliated to Qingdao University. 125 cases of normal lung tissue above 5 cm (the results of pathological tissue were diagnosed by the pathology department of Qingdao Municipal Hospital affiliated to Qingdao University). All the selected cases were not treated with chemotherapy and radiotherapy before operation. Immunohistochemical and Western Blot techniques were used to detect the expression of cystein-緯 -lyase in 125 cases of lung cancer and its adjacent normal mucosa, and followed up by out-patient visit or telephone consultation. The data of this study were analyzed by SPSS 13.0 software. The expression of cystathithione- 緯 -lyase in lung cancer tissues was significantly different from that in adjacent normal lung cancer tissues. Relationship between expression of cystathithione-緯-lyase and pathological and clinical factors in patients with lung cancer. Kaplan-Meier method was used to analyze the survival and draw the survival curve, and Log-rank was used to test its survival difference. Among them, P0.05 time indicates that the difference is statistically significant. Results: (1) cystathithione- 緯 -lyase was obviously expressed in lung cancer tissues, and the positive expression was mainly in the cytoplasm of lung cancer. The expression level of cystathithione- 緯 -lyase was significantly higher in lung cancer tissues than in adjacent normal tissues, and the expression level of cystathithione- 緯 -lyase in lung cancer tissues was significantly higher than that in adjacent normal tissues. There was significant difference in expression level between the two groups (P0.05). At the same time, Western blot Western Blot assay showed that the expression level of cystathithione- 緯 -lyase in lung cancer tissues was significantly higher than that in normal tissues adjacent to lung cancer. (2) the expression level of cystathion- 緯 -lyase in lung cancer tissues was similar to that in lung cancer tissues. The relationship between pathological and clinical factors and the 5-year survival rate of lung cancer patients were different: the expression of cystathion-gamma lyase and the TNM stage. The 5-year survival rate of lung cancer patients with low expression of cystathion- 緯-lyase was significantly higher than that of lung cancer patients with high expression of cystathion- 緯-lyase (P0.05). Conclusion: cystathithione- 緯 -lyase is highly expressed in lung cancer, which may play an important role in the development of lung cancer, invasion and metastasis. The expression of cystathithione- 緯 -lyase in lung cancer cells and the effect of cystathion- 緯 -lyase on proliferation and apoptosis of lung cancer cells will be helpful to understand the biological effects of cystathion- 緯 -lyase on lung cancer tumors. It may provide a certain degree of theoretical basis for the further research on cystathion-gamma lyase and lung cancer, and provide some strategies and means for the prevention and treatment of lung cancer. Therefore, the new idea of cystathithione- 緯 -lyase as a target may be used as a reference index to evaluate the early occurrence and prognosis of lung cancer.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R734.2

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