Soluble CD95L (s-CD95L) is a chemoattractant for certain lymphocyte subpopulations. We examined whether this ligand is a prognostic marker for high-grade serous ovarian cancer (HGSOC) and whether it is associated with accumulation of immune cells in the tumor. Serum s-CD95L levels in 51 patients with advanced ovarian cancer were tested by ELISA. IHC staining of CD3,
With the approval of new therapies targeting the PI3K pathway, the detection of PIK3CA mutations has become a key factor in treatment management for HR+/HER2− metastatic breast cancer (MBC). We developed multiplex digital PCR (dPCR) assays to detect and quantify PIK3CA mutations. A first screening assay allows the detection of 21 mutations, with a drop-off system targeting the 542–546 hotspot mutations combined with the simultaneous detection of N345K, C420R, H1047L and H1047R mutations. In the case of a positive result, a sequential strategy based on other assays that we have developped allows for precise mutation identification. Clinical validity was determined by analyzing plasma circulating free DNA (cfDNA) from 213 HR+/HER2− MBC samples, as well as DNA extracted from 97 available matched tumors from 89 patients. Our assays have shown reliable specificity, accuracy and reproducibility, with limits of blank of three and four droplets for the screening assay. Sixty-eight patients (32%) had at least one PIK3CA mutation detectable in their plasma, and we obtained 83.1% agreement between the cfDNA analysis and the corresponding tumors. The high sensitivity and robustness of these new dPCR assays make them well-suited for rapid and cost-effective detection of PIK3CA mutations in the plasma of MBC patients.
<p>Figure 1A: Membranous CD95 expression on ovarian cancer cell lines and primary cancer cells by cytometry Figure 1B: Evaluation of epithelial mesenchymal transition expression (EMT) profile using vimentin and E-cadherin expression on ovarian cancer cell line and primary cancer cells by Western Blot (A). Vimentin and E-cadherin expression ratio mean on ovarian cancer cells based on 3 experiments (B). Figure 1C: ALDH1 activity with or without s-CD95L (100 ng/mL) evaluated by cytometry. Figure 1D: Proliferation of OVCAR3 cell lines using cytometry and G1, G2 and S phase markers E: Proliferation of O170 primary cell lines using cytometry and G1, G2 and S phase markers F: Proliferation of various ovarian cancer cell lines or primary cell lines with or without s-CD95L (100 ng/mL) in medium using cytometry and G1, G2 and S phase markers</p>
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