PDF file - 1114K, S1. RT-PCR analysis illustrates the stable miR-106a expression in two benign (FTE187, T29) and malignant (SKOV3, HEY) cell lines.1 supplementary table S2. Growth curve of SKOV3 cell line with (red square) and without (blue diamonds) miR-106a overexpression in different doses of paclitaxel (A) and cisplatin (B)treatment in vitro. S3. Immunohistochemical stain of CD133 in four xenografts of SKOV3 tumor cells with (left panel) and without (right panel) miR-106a overexpression. S4. The side-population, major population of normal (T29) and malignant (HEY) cell lines with (top) and without (bottom) miR-106a overexpression were analyzed by flow cytometer. S5. Hematoxylin and eosin stained slides illustrate four xenografts of SKOV3 tumor masses without (left panel) and with (right panel) miR-106a overexpression. Supplementary Table 1 The published data show upregulation of miR-106a and its family members in ovarian cancer.
ARTICLE ABSTRACT
The degree of differentiation in human cancers generally reflects the degree of malignancy, with the most undifferentiated cancer being also the highest grade and the most aggressive. High-grade serous ovarian carcinomas (HGSOC) are poorly differentiated and fast-growing malignancies. The molecular mechanisms underlying the poor differentiation of HGSOC has not been completely characterized. Evidence suggests that miRNA, miR are dysregulated in HGSOC. Therefore, we focused on those miRNAs that are relevant to tumor differentiation. Expression profiling of miRNAs in HGSOC, indicated miR-106a and its family members were significantly upregulated. Upregulation of miR-106a was further validated by real-time reverse transcriptase PCR (qRT-PCR) and miRNA in situ hybridization in a large cohort of HGSOC specimens. Overexpression of miR-106a in benign and malignant ovarian cells significantly increased the cellular proliferation rate and expanded the side-population fraction. In particular, SKOV3 cells with miR-106a overexpression had significantly higher tumor initial/stem cell population (CD24- and CD133-positive cells) than control SKOV3 cells. Among many miR-106a predicated target genes, p130 (RBL2), an retinoblastoma (Rb) tumor suppressor family member, was not only confirmed as a specific target of miR-106a but also related to tumor growth and differentiation. The importance of mir-106a and RBL2 was further demonstrated in vivo, in which, SKOV3 cells overexpressing miR-106a formed poorly differentiated carcinomas and had reduced RBL2 levels. To our knowledge, this is the first study of miR-106a mediating proliferation and tumor differentiation in HGSOC.Implications: The current study suggests that the RB tumor suppressor pathway is a critical regulator of growth and differentiation in HGSOC. Mol Cancer Res; 11(11); 1314–25. ©2013 AACR.
