子宮內膜癌為已開發國家中最常見的婦科惡性腫瘤之一。目前已知腫瘤中ESR1的表現可以用於預測患者的存活率；然而，幹細胞因子如何影響子宮內膜癌的ESR1表現和對ESR1的標靶治療的反應仍然未知。本研究中，我們發現在子宮內膜癌中，幹細胞因子SOX2與ESR1之表現量呈負相關。更甚者，我們也發現高SOX2表現的患者具較差的存活率，而ESR1高表現的患者具較好的存活率，且兩者可同時用於預測子宮內膜癌患者之存活率。qPCR和immunoblot的結果顯示，子宮內膜癌細胞Ishikawa (ESR1陽性)及其後代Ishikawa-02 (ESR1陰性)除了分別具有較高和較低的SOX2表現外，兩者在細胞形態上具有明顯的差異。另外，我們也發現施予TGF-β會抑制SOX2的表現並增加ESR1的表現，因而誘導癌細胞塑性與細胞型態的改變。藉由clonogenic analysis，我們發現雖然Ishikawa、Ishikawa-02及其後代均對ESR1降解劑—Fulvestrant具有抗性，SOX2表現量較低的細胞卻會對CDK4/6抑制劑—Palbociclib較敏感。總括而言，我們發現了SOX2的表現量具有調控癌細胞可塑性的能力，而TGF-β刺激調控則對ESR1的表現量及子宮內膜癌細胞對標靶治療的敏感性有所影響。

Endometrial carcinoma is one of the most common gynecological cancer malignancies in developed countries, and ESR1 expression in tumors predicts survival outcome in patients. However, how stemness factors affect ESR1 expression and responses toward ESR1 targeted therapy in endometrial carcinoma is unknown. Here, we report that SOX2 stemness factor shows a negative expression pattern with ESR1 in endometrial carcinoma, and SOX2 and ESR1 expression predict a poor and good survival outcome, respectively, in patients. qPCR and immunoblotting analysis showed that endometrial carcinoma cell Ishikawa (ESR1-positive) and its descendent Ishikawa-02 (ESR1-negative) harbor low and high SOX2 expression, respectively, and display distinct morphology. We found that TGF-β stimulation downregulates SOX2, while upregulating ESR1 expression, and induces cancer cell plasticity with altered phenotypical changes in endometrial cancer cells. Clonogenic analysis showed that Ishikawa, Ishikawa-02, and their descendants are all resistant to fulvestrant, an ESR1 degrader, whereas SOX2-low cells are more sensitive to palbociclib, a CDK4/6 inhibitor. Our findings indicate the cancer cell plasticity mediated by SOX2 status and TGF-β stimulation affects ESR1 expression and responses toward targeted therapy in endometrial carcinoma.

目錄

Abstract ..........................................................................................................1

中文摘要............................................................................................................2

致謝...................................................................................................................3

Introduction.....................................................................................................7

Endometrial carcinoma..................................................................................7

SRY (sex determining region Y)-box 2 (SOX2) ............................................7

Estrogen receptor-1 (ESR1) ..........................................................................8

Hormone Therapy...........................................................................................9

Phosphatase and tensin homolog (PTEN) ................................................10

CDK4/6 inhibitor therapy..............................................................................10

Materials and Methods.................................................................................12

Cell Culture...................................................................................................12

Lentiviral Infection.......................................................................................12

Quantitative Real-Time PCR(Q-PCR) .........................................................13

Flow cytometry.............................................................................................13

Immunoblotting............................................................................................13

Clonogenic assay.........................................................................................14

Statistical Analysis.......................................................................................14

Result.............................................................................................................15

Cancer cell plasticity affects SOX2 and ESR1 expression pattern in endometrial carcinoma…….........................................................................15

Effect of TGF-β stimulation on SOX2 and ESR1 expression pattern in endometrial carcinoma……………………………….....................................15

SOX2 negatively regulates ESR1 expression............................................16

SOX2 and ESR1 as prognostic markers associated with survival in endometrial carcinoma patients.................................................................17

SOX2 status and TGF-β stimulation affect the effectiveness of targeted therapies………………………………............................................................17

Discussion....................................................................................................19

Figures..........................................................................................................20

References....................................................................................................30

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