Fig. 6: Downregulating neural lineage marker ARHGEF2 gene expression decreased C4-2B MDVR and H660 cell viability. | Communications Medicine

Fig. 6: Downregulating neural lineage marker ARHGEF2 gene expression decreased C4-2B MDVR and H660 cell viability.

From: Activation of neural lineage networks and ARHGEF2 in enzalutamide-resistant and neuroendocrine prostate cancer and association with patient outcomes

Fig. 6

a FPKM value of ARHGEF2, LHX2 and EPHB2 gene expression in 22RV1, LNCaP95, MSKCC-EF1 and H660 cells from GSE154576 dataset (n = 2). b Relative mRNA level of ARHGEF2, LHX2, and EPHB2 in C4-2B parental, MDVR, and H660 cells (n = 4). c Knocking down ARHGEF2 by siRNA decreased the expression of the markers of neuroendocrine including CHGA, NSE, and SYP in C4-2B MDVR and H660 cells. C4-2B MDVR and H660 cells were transfected with siRNAs for ARHGEF2 for 48 h, total cell lysates were collected and subjected to western blot analysis. d Knocking down ARHGEF2 by siRNA inhibited cell viability of C4-2B MDVR and H660 cells. C4-2B MDVR and H660 cells were transiently transfected with siRNAs targeting ARHGEF2 and viable cells were quantified using CellTiter Glo assay (n = 5). e qPCR analysis of AR targets and neuroendocrine markers and ARHGEF2 in NEPC LuCaP49 patient-derived xenograft (PDX) tumors and prostate adenocarcinoma LuCaP35CR tumors (n = 3). f Downregulation of ARHGEF2 by siRNA inhibits the growth of organoids derived from LuCaP49 PDX tumors. LuCaP49 organoids were seeded in a 96-well plate in the format of 3D Matrigel and then transfected with 50 nM ARHGEF2 siRNA and cultured for 14 days. The viability of the organoids was analyzed by CellTiter Glo and visualized by LIVE/DEAD staining (n = 4). Green = Calcein staining of live cells, Red = Ethidium homodimer-1 staining of dead cells. Scale bar 100 µm. *p < 0.05, **p < 0.01, ***p < 0.001 (mean and SD in a, b, d, e, f) using one-way ANOVA with multiple-comparisons test.

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