Non-coding RNA nc886 mediates TGF-β-induced mesothelial-to-mesenchymal transition

다국어 초록 (Multilingual Abstract)

Peritoneal dissemination is a common characteristic of ovarian cancer metastasis. Recently, an increasing number of studies have suggested that peritoneal mesothelial cells turn into carcinoma-associated fibroblasts through mesothelial-to-mesenchymal transition (MMT), which plays a role in ovarian cancer metastasis. However, molecular and cellular mechanism underlying the MMT in ovarian cancer microenvironment has not yet been elucidated. We found that nc886, which has been implicated in poor prognosis of patients with ovarian cancer, induced MMT resulting in the increased invasion of mesothelial cells. nc886 was enhanced in mesothelial cells stimulated by TGF-β which is highly secreted by tumor-associated macrophages. TGF-β-induced MMT and mesothelial cell invasion was significantly attenuated by nc886 knockdown. Knockdown of Smad4 significantly inhibited TGF-β-induced expression of nc886 and MMT makers. Notably, nc886 mediated the TGF-β-associated miR-200s expression in mesothelial cells. Moreover, miR-200b was associated with nc886-induced MMT and mesothelial cell invasion. Taken together, these results suggest that nc886 may play a key role in macrophage-induced ovarian cancer metastasis by mediating the TGF-β-induced MMT through the miR-200s/ZEB1.

목차 (Table of Contents)

• 1. ABSTRACT……………………………………………… 4
• 2. INTRODUCTION…………………………………………… 5
• 3. MATERIALS AND METHODS…………………………. 7
• 3.1. Materials…………………………….…….……… 7
• 3.2. Cell culture……… 7
• 3.3. Transfection…………………………………………. 8
• 3.4. Western blot …………………………….……………… 9
• 3.5. RT-PCR………………………………………………… 9
• 3.6. Invasion assay…………………………………………… 10
• 3.7. Statistical analysis……………………………………… 11
• 4. RESULT……………………………………………………. 14
• 4.1. nc886 induces the mesenchymal transition of mesothelial cells.………………………………………………. 14
• 4.2. nc886 is involved in TGF-β-induced MMT…………… 17
• 4.3. nc886 upregulation and MMT by TGF-β are associated with the Smad pathway…………………………………. 20
• 4.4. nc886 inhibits the expression of miR-200s regulated by TGF-β…………………………………………………. 22
• 4.5. nc886 promotes MMT and mesothelial cell invasion by regulating miR-200b…………………………………. 25
• 5. DISCUSSION…………………………………………. 29
• 6. REFERENCES…………………………………………. 33
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