探討ESM1調控FGFBP1參與人類子宮頸癌癌化和轉移之分子機轉

Exploring of the molecular mechanism of ESM1 modulating FGFBP1 expression involvement in tumorigenesis and metastasis of human cervical carcinoma

林怡安(Yi-An Lin)

指導教授 ： 謝逸憲

中山醫學大學/醫學院/醫學研究所/碩士(2023年)

https://doi.org/10.6834/csmu202300020

摘要

子宮頸癌是種能有效預防的癌症，但晚期子宮頸癌發生遠處轉移的現象是造成患者預後不佳最主要的原因，因此尋找子宮頸癌的生物標靶是目前醫療上重要的突破口。ESM1是由肺臟或腎臟內皮細胞所分泌的蛋白聚醣，目前研究指出ESM1於多種癌症中會高度表達，然而子宮頸癌和ESM1的關聯性仍未釐清，於是本次研究目的在探討ESM1於子宮頸癌中扮演的角色，並分析ESM1影響癌細胞的生物機轉。首先GEPIA資料庫顯示，高度表達ESM1之子宮頸癌患者整體存活率較低，且腫瘤的ESM1表現量高於正常組織；以西方墨點法和qPCR發現HeLa和SiHa細胞中ESM1表現量較高，ELISA實驗中也顯示相似的結果，於是將帶有shESM1質體之病毒液感染這兩株細胞以此抑制ESM1表達。後續發現抑制ESM1能有效降低子宮頸癌細胞生長及轉移能力，過度表達ESM1則誘導癌細胞增殖及轉移。透過RNA定序發現抑制ESM1會使FGFBP1基因表達下降，處理ESM1重組蛋白會使FGFBP1表達受到誘導。研究結果顯示抑制ESM1會使PKCα、FAK、Src和ERK磷酸化下降，並減少c-Jun和c-Fos的表達，利用免疫共沉澱發現FGFBP1會與ESM1相互結合。透過資料庫比對顯示FGFBP1 promoter上具有c-Jun的結合位點，推測ESM1透過c-Jun調控FGFBP1轉錄活性，以此調控癌細胞的生長及轉移。綜合以上研究結果，ESM1在調控子宮頸癌進程中扮演重要角色，未來還需探討更詳細的分子機制，期待ESM1成為新型標靶應用於子宮頸癌轉移治療。

關鍵字

子宮頸癌 ； 轉移 ； ESM1 ； FGFBP1 ； c-Jun

並列摘要

Cervical carcinoma is a largely preventable disease, but advanced cervical cancer is prone to distant metastasis, which is the main reason for the poor prognosis of patients. Therefore, looking for biomarkers in order to treat tumor progression is a therapeutic breakthrough point. Endothelial cell specific molecule 1 (ESM1) is mainly expressed in the endothelial cells in human lung and kidney tissues. Several studies have indicated that ESM1 is significantly overexpressed in many cancer types. However, the biological function and the mechanism of ESM1 in cervical carcinoma are still unknown. In this study, we found that the expression of ESM1 was significantly increased in the cervical tumor tissues and cervical cancer cell lines. Elevated ESM1 expression was significantly associated with an independent prognostic factor for the overall survival with patients of cervical cancer. Herein, we will explore the inhibition of human cervical cancer cells' metastasis and its related mechanism of ESM1. ESM1 knockdown showed that inhibit the proliferation, migration and invasion of HeLa and SiHa cells. Furthermore, we demonstrated that ESM1 knockdown significantly reduced the expression of phospho-ERK and phospho-FAK, which might participate in the regulation of cell metastasis. Treatment with human recombinant ESM1 protein induced the metastasis of HeLa and C33a cells in knockdown ESM1 cells. QPCR and western blot have confirmed that ESM1 knockdown can reduce the mRNA and protein expression of FGFBP1. In addition, ESM1 knockdown decreased the expression of c-Fos and c-Jun, the latter has a binding site on the promoter of FGFBP1. It is speculated that ESM1 modulates the transcriptional activity of FGFBP1 by decreasing the expression of c-Jun to regulate the proliferation and metastasis of cervical cancer cells. In conclusion, our study demonstrates the ESM1 may be a novel target for advanced cervical cancer patients and involved in tumorigenesis and metastasis of human cervical cancer cells, but detail molecular mechanism is need investigation in future.

並列關鍵字

Cervical cancer ； Metastasis ； ESM1 ； FGFBP1 ； c-Jun

參考文獻

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