Table_1_RETRACTED: LncRNA TCF7 Promotes Epithelial Ovarian Cancer Viability, Mobility and Stemness via Regulating ITGB8.docx

This study aimed to investigate the carcinogenic role of long non-coding RNA T-cell factor 7 (lnc-TCF7) in epithelial ovarian cancer (EOC). Lnc-TCF7 overexpression and shRNA plasmids were transfected into SKOV3 and OVCAR3 cells, followed by measurement of cell proliferation, migration, invasion, apoptosis, stemness, and mRNA profile (via microarray). Besides, lnc-TCF7 expression was measured in tumor and adjacent tissues from 76 EOC patients. Lnc-TCF7 was upregulated in EOC cell lines; its overexpression increased cell proliferation, migration, invasion, but decreased apoptosis and promoted CD44, CD133 expressions, CD44+CD133+ cell proportion, spheres formation efficiency and drug resistance to cisplatin in SKOV3 and OVCAR3 cells. Besides, lnc-TCF7 ShRNA exhibited opposite effects comparing with its overexpression. Microarray analysis revealed 267 mRNAs were modulated by lnc-TCF7 dysregulation, among which ITGB8 was the most dysregulated one, which was validated by subsequent western blot and RT-qPCR. Furthermore, ITGB8 overexpression not only induced proliferation, migration, invasion and stemness, but also attenuated the effect of lnc-TCF7 ShRNA on these functions in SKOV3 and OVCAR3 cells. In addition, lnc-TCF7 was upregulated in tumor tissues and correlated with higher pathological grade, tumor size, International Federation of Gynecology and Obstetrics (FIGO) stage and worse overall survival in EOC patients. Conclusively, lnc-TCF7 regulates multiple oncogenic pathways, promotes proliferation, migration, invasion, stemness via upregulating ITGB8. It also correlates with advanced tumor features and poor prognosis in EOC, implying its potential as a target for EOC treatment.

本研究旨在探讨长链非编码RNA T细胞因子7（long non-coding RNA T-cell factor 7，lnc-TCF7）在上皮性卵巢癌（epithelial ovarian cancer, EOC）中的致癌作用。将lnc-TCF7过表达质粒与短发夹RNA（short hairpin RNA, shRNA）质粒转染至SKOV3及OVCAR3细胞中，随后检测细胞增殖、迁移、侵袭、凋亡、干细胞特性，并通过基因芯片（microarray）分析mRNA表达谱。此外，本研究还对76例EOC患者的肿瘤组织及癌旁组织进行了lnc-TCF7的表达量检测。研究发现，lnc-TCF7在EOC细胞系中呈高表达；其过表达可增强SKOV3与OVCAR3细胞的增殖、迁移与侵袭能力，抑制细胞凋亡，并上调CD44、CD133的表达，提升CD44+CD133+细胞比例、球体形成效率及顺铂（cisplatin）耐药性。与之相反，lnc-TCF7的shRNA干扰可逆转上述效应。基因芯片分析显示，共有267个mRNA的表达受lnc-TCF7表达失调的调控，其中整合素β8（ITGB8）的表达失调最为显著，后续的蛋白质印迹（western blot）与实时定量聚合酶链反应（RT-qPCR）验证了这一结果。进一步研究表明，ITGB8过表达不仅可诱导细胞增殖、迁移、侵袭及干细胞特性，还可削弱lnc-TCF7 shRNA对SKOV3及OVCAR3细胞上述功能的调控作用。此外，lnc-TCF7在肿瘤组织中呈高表达，且与EOC患者的更高病理分级、更大肿瘤体积、国际妇产科联盟（International Federation of Gynecology and Obstetrics, FIGO）分期较晚及较差的总生存率显著相关。综上，lnc-TCF7可通过上调ITGB8调控多条致癌通路，促进细胞增殖、迁移、侵袭及干细胞特性；其还与EOC的进展期肿瘤特征及不良预后相关，提示其有望成为EOC治疗的潜在靶点。