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背景:绒毛膜细胞癌是一种具有高度侵袭能力的滋养细胞肿瘤。叉头蛋白D3作为胚胎干细胞和滋养干细胞的转录因子,在胚胎发生、肿瘤发生和进展等很多生理、病理状态下发挥着重要作用。目的:研究叉头蛋白D3在绒毛膜细胞癌恶性生物学特性中的作用及可能作用机制。方法:利用短发夹RNA干扰叉头蛋白D3在人绒毛膜细胞癌细胞株JAR中的表达,体外细胞增殖、迁移/侵袭实验检测JAR的细胞功能学改变,体内动物成瘤实验检测肿瘤生长情况。Agilent转录表达芯片初筛叉头蛋白D3调控下游基因并通过定量实时PCR验证。结果与结论:与原代培养的正常妊娠滋养细胞相比,绒癌细胞株JAR中叉头蛋白D3的转录和蛋白水平表达均显著增高。短发夹RNA干扰叉头蛋白D3表达抑制了体外JAR的细胞增殖、迁移和侵袭功能,同时抑制了裸鼠体内肿瘤的生长,降低了肿瘤细胞β-人绒毛膜促性腺激素分泌。通过初筛和后期验证,发现7个受叉头蛋白D3调控的局部黏附分子(ITGA5,ITGB6,THBS4,COL6A3,VTN,NRXN3和NLGN1),同时短发夹RNA干扰叉头蛋白D3表达后,JAR细胞内局部黏附激酶活性降低。提示过表达的叉头蛋白D3通过调控局部黏附分子表达和局部黏附激酶活性进而增强JAR的恶性生物学特性。
Background: Choriocarcinoma is a highly invasive trophoblastic tumor. As a transcription factor of embryonic stem cells and nourishing stem cells, forkhead protein D3 plays an important role in many physiological and pathological states such as embryogenesis, tumorigenesis and progression. Objective: To investigate the role of forkhead protein D3 in the malignant biological characteristics of human choriocarcinoma and its possible mechanism. Methods: Short hairpin RNA (shRNA) was used to interfere the expression of forkhead protein D3 (JAR) in human choriocarcinoma cell line JAR. Cell proliferation, migration and invasion were assayed in vitro. Cell viability was assayed in JAR and tumor growth in vivo . Primer with Agilent Transcriptional Expression Cholesterol D3 Regulates Downstream Genes and Validated by Quantitative Real-Time PCR. RESULTS AND CONCLUSION: Compared with the normal gestational trophoblast cultured in primary culture, the expression of forkhead protein D3 in JAR choriocarcinoma cell line was significantly increased. Short hairpin RNA interference expression of forkhead protein D3 inhibits the proliferation, migration and invasion of JAR cells in vitro, while inhibiting the growth of tumors in nude mice and reducing the β-human chorionic gonadotropin secretion in tumor cells. Seven primary adhesion molecules (ITGA5, ITGB6, THBS4, COL6A3, VTN, NRXN3 and NLGN1) regulated by the forkhead protein D3 were found by primary screening and later validation. Short hairpin RNA interfered the expression of the forkhead protein D3 Local intracellular adhesion kinase activity decreased. Tip overexpression of forkhead protein D3 through the regulation of local adhesion molecule expression and local adhesion kinase activity and thus enhance the malignant biological characteristics of JAR.