RESUMEN
Glypican-3 (GPC3) is overexpressed in hepatocellular carcinomas and hepatoblastomas and represents an important therapeutic target but the biologic importance of GPC3 in liver cancer is unclear. To date, there are limited data characterizing the biological implications of GPC3 knockout (KO) in liver cancers that intrinsically express this target. Here, we report on the development and characterization of GPC3-KO liver cancer cell lines and compare to them to parental lines. GPC3-KO variants were established in HepG2 and Hep3B liver cancer cell lines using a lentivirus-mediated CRISPR/Cas9 system. We assessed the effects of GPC3 deficiency on oncogenic properties in vitro and in murine xenograft models. Downstream cellular signaling pathway changes induced by GPC3 deficiency were examined by RNAseq and western blot. To confirm the usefulness of the models for GPC3-targeted drug development, we evaluated the target engagement of a GPC3-selective antibody, GC33, conjugated to the positron-emitting zirconium-89 (89Zr) in subcutaneous murine xenografts of wild type (WT) and KO liver cancer cell lines. Deletion of GPC3 significantly reduced liver cancer cell proliferation, migration, and invasion compared to the parental cell lines. Additionally, the tumor growth of GPC3-KO liver cancer xenografts was significantly slower compared with control xenografts. RNA sequencing analysis also showed GPC3-KO resulted in a reduction in the expression of genes associated with cell cycle regulation, invasion, and migration. Specifically, we observed the downregulation of components in the AKT/NFκB/WNT signaling pathways and of molecules related to cell cycle regulation with GPC3-KO. In contrast, pMAPK/ERK1/2 was upregulated, suggesting an adaptive compensatory response. KO lines demonstrated increased sensitivity to ERK (GDC09994), while AKT (MK2206) inhibition was more effective in WT lines. Using antibody-based positron emission tomography (immunoPET) imaging, we confirmed that 89Zr-GC33 accumulated exclusively in GPC3-expression xenografts but not in GPC3-KO xenografts with high tumor uptake and tumor-to-liver signal ratio. We show that GPC3-KO liver cancer cell lines exhibit decreased tumorigenicity and altered signaling pathways, including upregulated pMAPK/ERK1/2, compared to parental lines. Furthermore, we successfully distinguished between GPC3+ and GPC3- tumors using the GPC3-targeted immunoPET imaging agent, demonstrating the potential utility of these cell lines in facilitating GPC3-selective drug development.
RESUMEN
BACKGROUND: Submucosal myomas are associated with infertility and may be treated by hysteroscopic resection. OBJECTIVE: The aim of this retrospective study was to analyze 37 subfertile patients who unnderwent hysteroscopic myomectomy in a tertiary care center with particular regard to their postprocedure reproductive outcome. MATERIALS AND METHODS: The entire patient group (n = 37) underwent the procedure between March 2004 and March 2010. The submucosal myomas were type 0 (n = 27), type 1 (n = 8), and type 2 (n = 2). The mean myoma size was 2.1 cm; mean duration of the procedure was 54 mins and mean follow-up was 26 ± 10 months. 22 patients had one or more associated infertility factors. RESULTS: The complication rate was 5.4%. 11 patients (29.7%) conceived after the procedure. The pregnancy rate was better when myoma was the exclusive etiology of infertility (40%), when the myoma was completely intracavitary (33.3%), when the lesion was ≥ 30 mm in size (50%), and there were no associated intramural fibroids. CONCLUSION: Hysteroscopic myomectomy is a safe procedure to enhance fertility especially in cases with unexplained infertility.
RESUMEN
OBJECTIVE: To report a case of pseudoaneurysm causing massive hematuria with hemodynamic instability occurring after oocyte retrieval. DESIGN: Case report. SETTING: Tertiary care infertility clinic. PATIENT(S): A 34-year-old woman with endometriosis undergoing oocyte aspiration for IVF. INTERVENTION(S): Blood transfusion, cystoscopy, and resection and cauterization of pseudoaneurysm. MAIN OUTCOME MEASURE(S): Control of hematuria and hemodynamic stability. RESULT(S): Massive hematuria occurred after oocyte retrieval. Blood transfusion was given. Cystoscopy was done, and a bleeding pseudoaneurysm was seen. It was resected and cauterized. CONCLUSION(S): Pseudoaneurysm after oocyte retrieval has not been reported before. It resulted in massive hematuria causing hemodynamic instability. Immediate resuscitation and cystoscopy should be performed on all patients who present with persistent hematuria after oocyte retrieval. Injury to surrounding structures should always be kept in mind during oocyte retrieval.