尿液VEGF在診斷及追蹤人類攝護腺癌及膀胱癌之研究

A VEGF Biomarker Analysis of Urine for Diagnosis and Prognosis of Human Prostatic Carcinoma and Bladder Cancer

林朝欽(Tsao-Chin Lin)

指導教授 ： 林克亮

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2009年)

摘要

背景：惡性腫瘤在過去幾十年來一直位居台灣死亡排行榜上的首位。血管新生(angiogenesis)是腫瘤增生變大、延伸侵犯周邊組織、甚至於遠處轉移等腫瘤生長過程所需的條件。而血管內皮細胞生長因子(VEGF)則在血管新生及固態腫瘤扮演著重要角色，也是不可或缺的因子。本研究希望藉著收集不具侵襲性的尿液檢體並測定尿液中VEGF濃度來了解血管內皮生長因子分別在攝護腺癌及膀胱癌病人身上的相關性及重要性，以及其在致癌過程裡所扮演的角色，找出影響濃度變化與臨床特徵表現的關係，並藉著追蹤測定VEGF濃度之變化來比較血管內皮生長因子在攝護腺癌及膀胱癌病人治療前、中、後的影響，因而有機會用來評估腫瘤惡化的程度，以作為疾病預後之評估參考。
方法：本實驗收集攝護腺癌病例8位之檢體、膀胱癌病例10位之尿液檢體。另找11位正常成年人為陰性對照組。以市售商品套組(R&D systems)，利用Enzyme-Linked Immunosorbent Assay ( ELISA)測定VEGF評估作為攝護腺癌、膀胱癌的篩檢標記。與正常檢體的VEGF濃度做比較，然後再利用統計軟體分析其間的差異。
結果：本研究共計收集61人次檢體進行研究分析。測定尿液中VEGF protein的結果發現，攝護腺癌、膀胱癌患者VEGF濃度均高於沒有罹患任何癌症的人(p=0.001, p=0.012; Mann-Whitney檢定)。VEGF濃度在8位攝護腺癌患者的平均濃度為96.80 pg/ml、在10例膀胱癌患者的平均濃度為271.13 pg/ml ，另11位陰性對照組的VEGF值大都不高，平均濃度為2.60 pg/ml。攝護腺癌之格里森級數及膀胱癌分級與尿液VEGF濃度成正相關性，但個案太少(只有8例和10例)，顯示VEGF自我分泌能力強者，其疾病大多是比較惡化，是為一不好的疾病預後因子。
討論：本研究雖是初步的實驗研究，收集的病例不多，但仍提供了重要的訊息顯示VEGF在攝護腺癌、膀胱癌中亦是扮演重要角色。期望在後續的研究中，能收集單一特定疾病，做治療前、治療後VEGF值的檢測，以期能夠有效的提供疾病的預後評估及做為疾病治療的參考。這些VEGF腫瘤標誌非常具有潛力可以作為惡性疾病的診斷工具。

關鍵字

攝護腺癌 ； 膀胱癌 ； 血管內皮細胞生長因子

並列摘要

Background: Malignant tumors are at the top leading cause of death in Taiwan in the past decades. Angiogenesis is not only essential for tumor growth but is also implicated in invasion of the cancer cells into the circulation, and growth of dormant micro-metastases into frank metastatic lesions. Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis as well as in solid tumors. It also has a role for VEGF in Malignant tumors; although has not been fully elucidated. This study will examine the VEGF secretary activity of malignant cells in the patients with prostate cancer or bladder cancer. The urine samples were obtained before, during and after treatment. The purposes of this study are to assay the VEGF value by ELISA method and its correlation with disease prognosis in various prostate cancer or bladder cancer; and to evaluate the VEGF concentrations between prostate cancer and bladder cancer.

Method: The urine samples were collected from 8 prostate cancer and 10 bladder cancer patients in this study. It also was compared to 11 normal samples. An enzyme-linked immunosorbent assay (ELISA) was used to quantify VEGF concentrations. Urine samples will be tested using a commercial Elisa system for VEGF, (R&D systems). Identification of VEGF tumor markers for malignant disease using Enzyme-Linked Immunosorbent Assay technologies. The differential VEGF concentrations were statistically analyzed using bioinformatic softwares.

Results: Our research samples were 61 specimens. The VEGF value was low in 11 cases of Negative control(2.60 pg/ml), no obvious auto-secretary activity of those cells. In 8 cases of prostatic carcinoma and 10 cases of
bladder cancer, the VEGF urine level were 96.80 pg/ml(p=0.001) and
271.13 pg/ml(p=0.012) respectively( Mann-Whitney test). High VEGF level decreased significantly during treatment . However, the cases were too small to had exact predict value (only 8 and 10 cases). Patients were with high VEGF auto-secretary activity and also had bad prognosis.

Conclusion: Although our study is a primary result, study cases are varied, but it still provide important information that VEGF has an important role in prostate cancer and bladder cancer. We will process further research of single and specific disease in the future to analyze the exact correlation of VEGF and malignant tumor diseases before, during and after treatment. The VEGF tumor-specific markers show great potential for the detection of the Malignant tumors.

並列關鍵字

Prostatic Carcinoma ； Bladder Cancer ； VEGF

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