以酵素免疫分析法探討p90，p16及IMP1之自體抗體作為食道癌腫瘤標記的可行性

Study of autoantibody against to p90, p16 and IMP1 as tumor markers in human esophageal cancer by ELISA

張雅惠(Yea-Huey Chung)

指導教授 ： 陳凌雲

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2007年)

https://doi.org/10.6834/CSMU.2007.00082

摘要

食道癌為全世界第八大癌症。在台灣地區，所有癌症死亡原因中食道癌排名第九位。大多數的癌症患者到晚期才被確立診斷，往往造成預後不佳的情況，故食道癌病人早期的診斷具有重要的意義。有些學者採用SEREX方法，用癌症患者的血清尋找腫瘤抗原，有腫瘤特異性抗原(tumor-specific antigen, TSA)或腫瘤相關抗原(tumor- associated antigen, TAA)，這些物質含量的高低，可協助診斷癌症的變化或預後追蹤的工具。本實驗室王世吉學長以ELISA篩檢30位食道癌病患血清中IMP1、Cyclin B1、Ras、CSK、Erbb2、p90及p16七個腫瘤相關自體抗體的表現，結果陽性率較高的有p90為10%，p16為26.6%，IMP1為26.6%，似乎p90、p16及IMP1可能可以作為診斷食道癌的腫瘤標幟物，為了進一步確認，本實驗將食道癌病患的檢體數目從30位增加到120位，本實驗除了用原液血清之外，也用硫酸銨沉澱血清蛋白以提高敏感度，並以兩個不同的cutoff value，分別為正常的cutoff value (Mean+2SD)及較低的cutoff value (Mean+SD)，來比較正常人及病人的陽性率，並比較原液及硫酸銨血清之差異。實驗結果顯示，以正常的cutoff value (Mean+2SD)所得之p90、p16與IMP1自體抗體的陽性率，在正常人原液血清分別為3.33%、0%、0%，在病人原液血清中分別為3.33%、5.00%及8.33%。而以較低的cutoff value (Mean+SD)分析p90、p16與IMP1在正常人中分別為23.33%、23.33%、20.00%，在病人為9.17%、14.17%、21.67%。明顯發現cutoff value降低時，會使陽性率增加但準確度降低。而硫酸銨處理的血清，如以正常的cutoff value (Mean+2SD)分析p90、p16與IMP1的陽性率，在正常人分別為0%、0%、3.33%，在病人則為3.33%、5.83%、10.00%；若以較低的cutoff value (Mean+SD)分析，則正常人為16.67%、20.00%、20.00%，在病人則為10.00%、15.83%、25.00%。亦明顯發現cutoff value降低時，會使陽性率增加但準確度降低。我們為了增加陽性率的準確度，我們進一步交叉分析所有結果，發現經硫酸銨沉澱的血清，有自體抗體p90或p16表現的正常人或病人中，在正常人中只有10%會同時表現p90與p16，但在病人中則有高達50%同時表現p90與p16。顯示能以p90與p16同時存在來篩選食道癌罹患之可能性。此結果，在原液血清中則不存在。

關鍵字

食道癌 ； 腫瘤標記

並列摘要

Esophageal cancer is the eighth major cancer worldwide. Esophageal cancer ranks the ninth cause of death from all cancers in Taiwan. Most cancer patients were not established to diagnose until later period. They were in the situation with poor prognosis. Therefore, it is important in the early diagnosis of esophageal cancer patients. Some scholars were looking for the tumor antigens with the cancer patients' sera by SEREX. The level of tumor antigens which are tumor-specific antigens or tumor-associated antigens can help diagnose the change or prognosis of the cancer as the tracked tool. The person who we went to school with in our laboratory had screened seven autoantibodies against TAAs including IMP1, Cyclin B1, Ras, CSK, Erbb2, p90, p16 in 30 esophageal cancer patients’ sera by ELISA. Three autontibody to p90, p16 or IMP1 presents in high positive reactors. The percentages of positive reactors to p90, p16 or IMP1 were higher relatively. The percentages were 10%, 26.6% and 26.6% respectively. It seems that p90, p16 and IMP1 might be diagnosed as tumor markers of the esophageal cancer. Because the specimens are too little, we increase from 30 to 120 specimens to confirm if p90, p16 and IMP1 are useful tumor markers in esophageal cancer. We use both original sera and sera treated with ammonium sulfate that precipitates the serum proteins by improving sensitivity. Two different cutoff values are normal cutoff value (Mean +2SD) and lower cutoff value (Mean +SD) respectively. We compare the percentage of positive reactors in patients and healthy individuals. And we contrast between original sera and sera from ammonium sulfate. By normal cutoff value (Mean +2SD), the results show that autoantibody to p90, p16 and IMP1 were found in 3.33%, 0%, 0% of healthy individuals’ and 3.33%, 5.00%, 8.33% of patients’ original sera respectively. And the lower cutoff value (Mean +SD) analyses p90, p16 and IMP1. They appear 23.33% , 23.33% , 20.00% in healthy individuals and 9.17%, 14.17%, 21.67% in patients respectively. We find that lower cutoff value increases percentages of positive reactors but decreases the accuracy obviously. And normal cutoff value (Mean+2SD) analyses p90, p16 or IMP1 in sera treated with the ammonium sulfate. They appear 0%, 0%, 3.33% in healthy individuals and 3.33%, 5.83%, 10.00% in patients respectively. By lower cutoff value (Mean +SD), percentage of positive reactors are 16.67%, 20.00%, 20.00% in healthy individuals and 10.00%, 15.83%, 25.00% in patients. We also find that lower cutoff value increases the percentages of positive reactors but decreases the accuracy obviously. In order to increase the accuracy of positive reactors, we cross the all results. We find the sera treated with ammonium sulfate. Among the healthy individuals or patients appear p90 or p16 autoantibody, p90 and p16 which appear at the same time are 10% in healthy individuals and 50% in patients. It is possible that both of p90 and p16 appear at the same time to screen the esophageal cancer. These results are absent in the original sera.

並列關鍵字

esophageal cancer ； tumor marker

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