Project description:Evaluation of X-linked gene expression in MEFs isolated from STC1 wild type and knock out mice X chromosome inactivation (XCI) is initiated in cis by the Xist RNA, which coats the inactive X chromosome (Xi) from which it is produced. We performed a large-scale RNA interference screen to identify trans-acting XCI factors (XCIFs) that comprise regulators of cell signaling and transcription. We find that the XCIFs promote Xist expression and/or localization to the Xi. One of the XCIFs, STC1, is a glycoprotein found in both the cytoplasm and nucleus and whose function is poorly understood. A homozygous mouse knockout of STC1 has a defect in XCI but surprisingly is phenotypically normal. We performed transcriptome profiling (RNA-Seq) experiments to determine whether the expression levels of X-encoded genes were elevated in Stc1-/- female MEFs. In these experiments, RNA was prepared from three independent cultures of Stc1+/+ or Stc1-/- female MEFs. RNA samples were processed and amplified followed by deep sequencing. The similarity of X-linked gene expression between Stc1+/+ and Stc1-/- MEFs was statistically significant. The vast majority of autosomal genes were also expressed at comparable levels in Stc1+/+ and Stc1-/- MEFs. Sequenced mRNA isolated from the STC1+/+ or STC1-/- MEFs.
Project description:Evaluation of X-linked gene expression in MEFs isolated from STC1 wild type and knock out mice X chromosome inactivation (XCI) is initiated in cis by the Xist RNA, which coats the inactive X chromosome (Xi) from which it is produced. We performed a large-scale RNA interference screen to identify trans-acting XCI factors (XCIFs) that comprise regulators of cell signaling and transcription. We find that the XCIFs promote Xist expression and/or localization to the Xi. One of the XCIFs, STC1, is a glycoprotein found in both the cytoplasm and nucleus and whose function is poorly understood. A homozygous mouse knockout of STC1 has a defect in XCI but surprisingly is phenotypically normal. We performed transcriptome profiling (RNA-Seq) experiments to determine whether the expression levels of X-encoded genes were elevated in Stc1-/- female MEFs. In these experiments, RNA was prepared from three independent cultures of Stc1+/+ or Stc1-/- female MEFs. RNA samples were processed and amplified followed by deep sequencing. The similarity of X-linked gene expression between Stc1+/+ and Stc1-/- MEFs was statistically significant. The vast majority of autosomal genes were also expressed at comparable levels in Stc1+/+ and Stc1-/- MEFs.
Project description:identify the potential partners of STC1 at the protein level, we performed mass spectrometry on B16-F10 tumor cells stably expressing FLAG-tagged STC1.
Project description:Resistance to temozolomide (TMZ) is one of the major challenges for glioblastoma (GBM) therapy while the underlying mechanisms demand further exploration. Tumor-repopulating cells (TRCs) have been proven to be involved in chemotherapy resistentce. We first enriched GBM TRCs by culturing DBTRG cells in 3D soft fibrin gels and performed RNA-seq. By anlyzing the differentially expressed genes (DEGs) between TRCs and 2D conventionally cultured DBTRG cells, we identified the glycoprotein gene Stanniocalcin-1 (STC1), which is highly expressed in TRCs. Our analyses using patient data from CGGA disclosed that high STC1 expression was associated with poor prognosis, high glioma grade and TMZ therapy resistance. Both our in vitro and in vivo expreriments showed that overexpression of STC1 promoted while knockdown of STC1 inhibited GBM cell proliferation and TMZ resistance. Our mechanistic study revealed that STC1 elevated the phosphoralation of STAT3 to increase MGMT expression, which inhibited the TMZ-induced DNA damage and apoptosis. Our study provides rationale for targeting STC1 to overcome TMZ resistance.
Project description:Analysis of genes regulated by STC1 down-regulation in mouse 4T1 derived clone, 4T1ch9. STC1 expression is associated with tumor growth and metastasis. This study looks at genes affected when STC1 expression is down-regulated by STC1 shRNA.
Project description:Analysis of genes regulated by STC1 down-regulation in mouse 4T1 derived clone, 4T1ch9. STC1 expression is associated with tumor growth and metastasis. This study looks at genes affected when STC1 expression is down-regulated by STC1 shRNA. Total RNA isolated from shRNA transduced cells subjected to puromycin selection for 5-6 days.
Project description:Stanniocalcin 1 (STC1) plays an integral role in the metastasis of ovarian cancer. However, the functional role of STC1 in lipid metabolism is not fully understood. Single-cell sequencing assays verified that STC1 expression was significantly enhanced in ovarian cancer tissues compared with para-carcinoma tissues, and it was further up-regulated in peritoneal metastasis tissues compared with tumor tissues. In vitro and in vivo experiments demonstrated that STC1 promoted cell proliferation and metastasis by enhancing lipid metabolism. Mechanistically, STC1 directly bound to integrin β6 (ITGB6) and activated the PI3K signaling pathway. Moreover, STC1 was directly regulated by FOXC2 and FOXC2 was up-regulated and positively correlated with STC1 in ovarian cancer. Notably, STC1 knockdown had a synergistic effect with cisplatin (DDP) chemotherapy.Overall, these findings reveal that STC1 increases metastasis by promoting lipid metabolism via the FOXC2/ITGB6 signaling axis and may be a potential target for chemotherapy-resistant ovarian cancer.
Project description:We sequenced and analyzed the genome of a highly inbred miniature Chinese pig strain, the Banna Minipig Inbred Line (BMI). we conducted whole genome screening using next generation sequencing (NGS) technology and performed SNP calling using Sus Scrofa genome assembly Sscrofa11.1.