Stem and Progenitor-Like Cells Contribute to the Aggressive Behavior of Human Epithelial Ovarian Cancer

Stem and Progenitor-Like Cells Contribute to the Aggressive Behavior of Human Epithelial Ovarian Cancer. shorter PFS and OS reaching borderline significance (= 0.06; = 0.07). LIN-28-and SOX-2 positive cells were detected in all eight pts AT. Patients and Methods 79 POC pts were studied for DTCs before therapy (BT) and after therapy (AT) using immunocytochemistry. Eight pts harboring at least five DTCs AT were further analyzed on two additional slides by four-fold immunofluorescence staining for DAPI, Cytokeratin (CK), SOX-2 or LIN-28, CD45 and CD34 (Cy5). A stem-like tumor cell was classified as Dapipos, CD45neg, CD34neg, SOX-2pos/LIN-28pos and CKpos or CKneg. Conclusions Crizotinib hydrochloride Stem cell associated proteins are expressed in DTCs that are present AT and their presence seem to be correlated with a worse outcome. Additional therapeutic regimens may be necessary to eliminate these cells. = 0.02) and patients initially DTCneg BT but DTCpos AT had a significant shorter PFS (= 0.03) (Table ?(Table22 and Physique ?Physique1).1). The persistence of DTCs resulted in a shorter PFS and OS reaching borderline significance (= 0.06; = 0.07). Open in a separate window Physique 1 Kaplan-Meier analysis for the correlation of PFS (ACD) and OS (ECH) with DTC detectionPatients initially DTCneg before therapy but DTCpos after therapy had a significant shorter PFS (= 0.03) (Physique ?(Figure1D).1D). A. PFS DTCpos/pos, B. PFS DTneg/neg, C. PFS DTCpos/neg, D. PFS DTCneg/pos, E. OS DTCpos/pos, F. OS DTCneg/neg, G. OS DTCpos/neg, H. OS DTCneg/pos. Evaluation of LIN28- and SOX-2-positive cells Staining of patient samples is shown in Figures ?Figures22C5. Controls are shown in Supplementary Figures S1CS3. A DTC was classified as a stem-like tumor cell if it had the following staining characteristics: Dapipos, CD45neg, CD34neg, SOX-2pos/LIN-28pos and CKpos or CKneg (Figures ?(Figures22C5). The Kasumi cell line was used to establish CD34 expression (Supplementary Physique 1) and BM samples from healthy donor patients for CD34- and CD45-expression (Supplementary Figures 2 and 3). Open in a separate window Physique 2 Representative four-fold immunofluorescence staining for CKpos/LIN-28pos cells after therapy of patient No1(A) Cell nuclei were stained with Dapi. (B) Indicates a CKpos cell. (C) Crizotinib hydrochloride Alludes a cell with LIN-28pos phenotype. (D) Shows CD34pos and/or CD45pos cells. (E) Indicates a merge of a DTC with the phenotype Dapipos, CKpos, LIN-28pos, CD34neg and CD45neg, magnification at 63. Open in a separate window Physique 5 Representative four-fold immunofluorescence staining for CKneg/SOX-2pos cells after therapy of patient No1(A) Cell nuclei were stained with Dapi. (B) Indicates a CKneg cell. (C) Alludes a cell with SOX-2pos phenotype. (D) Shows CD34pos and/or CD45pos cells. (E) Indicates a merge of a DTC with the phenotype Dapipos, CKneg, SOX-2pos, CD34neg and CD45neg, magnification at 63. Open in a separate window Physique 3 Representative four-fold immunofluorescence staining for CKneg/LIN-28pos cells after therapy of patient No1(A) Cell nuclei were stained with Dapi. (B) Indicates a CKneg cell. (C) Alludes a cell with LIN-28pos phenotype. (D) Shows CD34pos and/or CD45pos cells. (E) Indicates a merge of two DTCs with the phenotype Dapipos, CKneg, LIN-28pos, CD34neg and/or CD45neg, magnification at 63. Open in a separate window Physique 4 Representative four-fold immunofluorescence staining for CKpos/SOX-2pos cells after therapy of patient No1(A) Cell nuclei were stained with Dapi. (B) Indicates a CKpos cell. (C) Alludes a cell with SOX-2pos phenotype. (D) Shows CD34pos and/or CD45pos cells. (E) Indicates a merge of a DTC with the phenotype Dapipos, CKpos, SOX-2pos, CD34neg and/or CD45neg, magnification at 63. Detection of LIN-28- and SOX-2-positive cells DTCs from 10 patients were analyzed BT and AT for SOX-2 and LIN-28 positive cells AT (Table ?(Table3;3; columns 1 and 2; Supplementary Table 1). 8/10 patients had at least five DTCs as detected by immunocytochemistry using A45/B-B3. In addition, 2/10 patients (patient 2 and 5) were DTCneg AT but DTCposBT. As apparent from Table ?Table3,3, Crizotinib hydrochloride AT CKpos/LIN-28pos cells were detected in 9/10 patients [median 2 cells (range 1C5)] and CKneg/LIN-28pos cells in 7/10 patients [median 3 cells (range 1C11)], respectively. CKpos/SOX-2pos cells were detected in 6/8 patients [median 2 cells (range-0C4)] and CKneg/SOX-2pos cells were found in 7/8 patients [median 4 cells (range 1C11)]. Patients two and five, who were characterized as DTCneg AT by immunocytochemistry but were positive BT (37 and 6 DTCs, respectively) were included in our analysis for stem cell-associated markers. Interestingly, these two patients harbored 1C2 LIN-28pos and SOX-2pos Crizotinib hydrochloride cells in their BM AT. Thus, we evaluated LIN-28-/SOX-2-positive cells BT in cases vice versa, DTCneg BT but DTCpos AT (patients 1, 3 and 4) as well as in patient number 6 6 with persistent DTCs. As shown in Table ?Table3,3, in patients who switched initially DTCneg before but Rabbit polyclonal to MTH1 becoming DTCpos AT (patients 1, 3 and 4), in patients who switched from.