藉由蛋白質體學分析子宮頸癌 與轉移相關之生物標記分子

根據最新台灣衛生署102年統計，國人死大死因榜首為惡性腫瘤，而子宮頸癌位居於女性癌症中第七位。而根據WHO之2011年統計子宮頸癌為女性癌症中發生率第三高之癌症，每年約有530,000新診斷子宮頸癌之個案，且一半會因此而死亡。癌症死亡率如此高之原因主要由於癌症組織除了會在原部位發展外，到晚期會進行遠端轉移至其他維生器官如肝、腦、肺等等，並極有可能造成病患死亡。而子宮頸癌早期之五年存活率相當高，但當轉移一旦發生時，五年存活率變大幅降低。因此，在本論文研究中將具有不同轉移能力之子宮頸癌細胞株利用蛋白質體學之方式，探討轉移相關機制以及找到轉移相關之生物標記分子。在本實驗中我們利用相同基因背景之HeLa與較具有侵襲能力之HeLa-I5做研究並希望找出具有潛力之轉移相關之蛋白。我們利用二維差異電泳(2D-DIGE)、基質輔助雷射脫附游離飛行時間質譜儀(MALDI-TOF MS)找出差異之蛋白質。在結果中共鑑定出68個蛋白質具有差異，並利用西方點墨法(western blot)驗證。在這些差異蛋白質中，我們選出一個較有潛力之蛋白-progesterone receptor membrane component1(PGRMC1)做後續實驗。並利用核醣核酸干擾技術(RNAi)使PGRMC1基因沉默。我們利用Transwell migration、傷口癒合實驗來觀察其蛋白對細胞移動能力之影響；Transwell invasion觀察細胞對細胞侵襲能力之影響，驗結果可以得知，PGRMC1對於子宮頸癌細胞之移動、侵襲能力以及對於細胞增生的速度有正面影響。本實驗利用蛋白質體學分析法找出在子宮頸癌轉移具有差異之蛋白質，並找出許多參與癌細胞侵入與轉移相關之潛力蛋白如PGRMC1，並希望能提供子宮頸癌與轉移相關的生物標記分子。

關鍵字

轉移；子宮頸癌； PGRMC1 ；蛋白質體學

並列摘要

In Taiwan, Cervical cancer (CC) is the 7th leading cause of death in female cancer. Moreover, according to the WHO's report in 2011, cervical cancer is the third highest cancer incidence in female, around 530,000 new cases of cervical cancer were diagnosed each year with high death rate. The reason for a high mortality of cervical cancer is mainly due to cancer metastasis. When cervical cancer is diagnosed in early stage, the five-year survival rate is very high, but once metastasis occurs, the five-year survival rate reduced significantly. Thus, to manage cervical cancer, finding metastasis biomarkers and understanding the mechanisms are critical. In our current study, we used a pair of cervical adenocarcinoma cell lines, HeLa and its invasive partner, HeLa-I5, as a model system to examine invasive mechanism and find out the potential cellular targets associated with metastasis. We use two-dimensional differential gel electrophoresis difference (2D-DIGE), matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) to identify differentially expressed proteins between HeLa and HeLa-I5. Our proteomics study revealed that 68 proteins show significant difference between the two lines and some of the identified proteins have been validated through immunoblotting. Further study by using RNA interference (RNAi) technique, we found progesterone receptor membrane component1 (PGRMC1) protein is a key player in response to cervical cancer invasion, migration and proliferation. Our data demonstrated that the invasive ability and the migration ability decreased for 2-fold and 1.7-fold, respectively, in the process of PGRMC1 knockdown. To sum up, PGRMC1 has been evidenced to modulate cervical cancer migration, invasion and proliferation. Our data also provided useful diagnostic markers and therapeutic candidates for the treatment of cervical cancer invasion and migration.

並列關鍵字

metastasis ； cervical cancer ； PGRMC1 ； proteomics

參考文獻

2 Williams, N. L., Werner, T. L., Jarboe, E. A. & Gaffney, D. K. Adenocarcinoma of the cervix: should we treat it differently? Current oncology reports 17, 17, doi:10.1007/s11912-015-0440-6 (2015).

3 Freire-de-Lima, L. Sweet and sour: the impact of differential glycosylation in cancer cells undergoing epithelial-mesenchymal transition. Frontiers in oncology 4, 59, doi:10.3389/fonc.2014.00059 (2014).

4 Yates, J. R., 3rd. Mass spectrometry and the age of the proteome. Journal of mass spectrometry : JMS 33, 1-19, doi:10.1002/(SICI)1096-9888(199801)33:1<1::AID-JMS624>3.0.CO;2-9 (1998).

5 Lander, E. S. et al. Initial sequencing and analysis of the human genome. Nature 409, 860-921, doi:10.1038/35057062 (2001).

6 Venter, J. C. et al. The sequence of the human genome. Science 291, 1304-1351, doi:10.1126/science.1058040 (2001).

國際替代計量

藉由蛋白質體學分析子宮頸癌與轉移相關之生物標記分子

全文下載

主題瀏覽