復發性大腸直腸癌診斷標誌暨診斷晶片之研發

Molecular Markers and Development of Diagnostic Multigene Biochip for Relapsed Colorectal Cancer

張鈺堂(Yu-Tang Chang)

指導教授 ： 王照元

高雄醫學大學/醫學院/醫學研究所博士班/博士(2018年)

摘要

本研究主要以大腸直腸癌患者癌細胞所具有之基因作為候選生物標誌，利用加權酵素晶片陣列平台 (Weighted Enzymatic Chip Array Platform)，針對大腸直腸癌患者的周邊血液進行追蹤檢測，研發高效能檢測早期復發 (術後一年內) 與復發的大腸直腸癌患者周邊血中循環腫瘤細胞相關生物標誌晶片。我們以在大腸直腸癌患者癌組織中過度表現的十九個mRNA多基因生物晶片，確認能比傳統血清CEA更有效預測大腸直腸癌患者的手術後復發。接著以不同的組合排列，結果顯示ELAVL4、PTTG1、BIRC5、PDE6D、CHRNB1、MMP13、PSG2七個基因標誌是最佳組合，所建構的多基因生物標誌晶片，針對早期復發的預測，接收者操作特徵曲線 (receiver operating characteristic curve；ROC curve) 下面積 (Area under the Curve of ROC；AUC) 為0.854，準確度為83.2%；而針對復發的預測，AUC為0.884，準確度為88.2%。七個標誌組合的生物晶片，可提早3.8個月預測早期復發，提早10.4個月預測復發。相較於傳統以單一腫瘤標誌為檢測標的方式，此生物標誌晶片顯著提升了檢測效度。晶片陽性的五年整體存活率為33.7%，陰性五年整體存活率為95.6% (P < 0.001)；晶片陽性的五年無病存活率為30.9%，陰性為91.9% (P < 0.001)。本研究結果顯示，利用多基因生物晶片檢測血液循環生物標誌比傳統血清CEA可更有效預估大腸直腸癌患者的早期復發與復發，晶片上基因標誌的組合與其對於患者復發檢測的能力顯著相關，且個別生物標誌的表現亦可同時提供患者預後狀況的評估。

關鍵字

分子偵測 ； 大腸癌 ； 術後早期復發 ； 術後復發 ； 循環腫瘤細胞

並列摘要

We previously constructed a multigene biochip with 19 candidate genes. Compared with conventional serum carcinoembryonic antigen detection, our multigene chip aided more accurate and earlier prediction of postoperative relapse during stage I-III colorectal cancer (CRC) patient surveillance. The second study was to optimize a multigene biochip for detecting the risk of postoperative early relapse and relapse in patients with CRC. Of the 19 circulating biomarkers, ELAVL4, PTTG1, BIRC5, PDE6D, CHRNB1, MMP13, and PSG2, which presented significant predictive validity, were selected for combination. The expression of the 7-biomarker biochip resulted in area under the receiver operating characteristic curve values of 0.854 (95% confidence interval [CI]: 0.756-0.952) for early relapse and 0.884 (95% CI: 0.830-0.939) for relapse. The median lead times prior to the detection of postoperative early relapse and relapse were 3.8 and 10.4 months, respectively. From 19 circulating biomarkers, we optimized 7 contemporary circulating biomarkers. The prediction model used for the early and accurate identification of Taiwanese patients with CRC having a high risk of postoperative early relapse and relapse seems to be feasible and comparable.

並列關鍵字

molecular detection ； relapse ； early relapse ； circulating tumor cell

參考文獻

Allard WJ, Matera J, Miller MC, Repollet M, Connelly MC, Rao C, Tibbe AG, Uhr JW, Terstappen LW. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004; 10: 6897-6904.

Almog N. Molecular mechanisms underlying tumor dormancy. Cancer Lett. 2010; 294: 139-146.

Alunni-Fabbroni M and Sandri MT. Circulating tumour cells in clinical practice: methods of detection and possible characterization. Methods. 2010; 50: 289-297.

Amado RG, Wolf M, Peeters M, Van Cutsem E, Siena S, Freeman DJ, Juan T, Sikorski R, Suggs S, Radinsky R, Patterson SD, Chang DD. Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J Clin Oncol. 2008; 26: 1626-1634.

André T, Boni C, Navarro M, Tabernero J, Hickish T, Topham C, Bonetti A, Clingan P, Bridgewater J, Rivera F, de Gramont A. Improved overall survival with oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment in stage II or III colon cancer in the MOSAIC trial. J Clin Oncol. 2009; 27: 3109-3116.