Journal: Cell death & disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism.

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: Fig. 4 Downregulation of CHKα mediates the inhibitory effect of DHRS2 on the growth and invasion of OC cells. A The protein expression levels of CHKα in the designated group. After treated with 10 μM choline for 3 days or transfection with CHKα, B the viability of the cells was detected by MTS, C the invasive capability of the cells was determined by cell invasion assay, and D the protein levels of p-AKT and perilipin1 were detected by Western blotting. E–G After treated with actinomycin D (5 μg/ml), CHKα mRNA levels were measured by RT-qPCR, and the percentage of remaining mRNAs in the designated group were plotted. H RIP assay showing that DHRS2 can bind to CHKα mRNA in both OVCAR3 and HO-8910 cells. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences (p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The antibodies to detect CHKα (13520-1-AP, 1:500), AKT (10176-2-AP, 1:1000), and phospho-AKT (S473) (66444-1-Ig, 1:1000) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Expressing, Transfection, Invasion Assay, Western Blot, Quantitative RT-PCR

Journal: Cell death & disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism.

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: Fig. 5 DHRS2 impairs tumor growth in vivo by interfering with the CHKα-AKT axis and choline metabolism. A The growth curve of OVCAR3-CON and OVCAR3-DHRS2 cells in vivo. Female BALB/c nu/nu mice were subcutaneously inoculated with OVCAR3-CON and OVCAR3- DHRS2 cells (n = 6 per group). Tumor volume was examined every other day and shown in the graph. B At the end of the experiment, the mice were sacrificed and the tumors were separated. C Tumor mass of each group was measured and shown in the graph. D 11C-Choline micro-PET/CT was performed and standardized uptake value (SUV) intensity was observed in the designated groups. E Representative images of Oil Red O staining in tumor tissues of the designated group. F The protein levels of DHRS2, CHKα, p-AKT, AKT, and perilipin1 in tumor tissues of the designated group. G Representative images of tumor sections in each group stained with indicated antibodies. Antibody staining is in brown and nuclear counter staining is in blue. Scatter diagram shows Histoscore for the indicated antibody staining in tumor samples. Survival analysis from TCGA dataset assessed by the Kaplan–Meier method for OC patients with high or low signature of H DHRS2 and I CHKα genes. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences (p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The antibodies to detect CHKα (13520-1-AP, 1:500), AKT (10176-2-AP, 1:1000), and phospho-AKT (S473) (66444-1-Ig, 1:1000) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: In Vivo, Micro-PET, Staining

Journal: Cell death & disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism.

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: Fig. 6 DHRS2 hampers the invasion and metastasis of OC cells. A Representative images of spheroids formed by OVCAR3 cells in Matrigel (left). The invasive spheroids were featured by scattered protrusions formed on the spheroid surface, as indicated by yellow arrows. Scale bar, 50 μm. Proportion of two spheroid types in OVCAR3-CON and OVCAR3-DHRS2 cells (right) (p < 0.05). Stable OVCAR3-CON and OVCAR3-DHRS2 cells were injected into the abdominal cavity, respectively (n = 6 per group). B During the experiment, the body weight of the mice was determined every 3 days and shown in the graph. C Representative intraperitoneal metastatic tumors in the designated group. D, E The amount and mass of the metastatic tumors were shown in the graph. F The protein levels of DHRS2, CHKα, p-AKT, and perilipin1 in tumor tissues of the designated group. G Representative images of tumor sections in each group stained with indicated antibodies. H The representative images of DHRS2 staining in OC tissues and non-tumor tissues. The scatter diagram shows Histoscore for DHRS2 staining in the designated samples. Data are shown as mean values ± S.D. of independent, triplicate experiments. The asterisks (*,**,***) indicate significant differences (p < 0.05, p < 0.01, p < 0.001, respectively).

Article Snippet: The antibodies to detect CHKα (13520-1-AP, 1:500), AKT (10176-2-AP, 1:1000), and phospho-AKT (S473) (66444-1-Ig, 1:1000) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Injection, Staining

Journal: Cell death & disease

Article Title: DHRS2 inhibits cell growth and metastasis in ovarian cancer by downregulation of CHKα to disrupt choline metabolism.

doi: 10.1038/s41419-022-05291-w

Figure Lengend Snippet: Fig. 7 Schematic illustration of DHRS2 inhibiting OC growth and invasion by regulating choline metabolism. Left, CHKα mRNA is translated to protein, which promotes the activation of choline metabolism and AKT pathway, thereby promoting the growth and invasion of OC. Right, DHRS2 can directly bind to CHKα mRNA to accelerate its degradation. Correspondingly, CHKα protein expres- sion is downregulated, resulting in the disruption of choline metabolism and AKT signaling. These effects lead to the inhibition of OC growth and metastasis. OC, ovarian cancer.

Article Snippet: The antibodies to detect CHKα (13520-1-AP, 1:500), AKT (10176-2-AP, 1:1000), and phospho-AKT (S473) (66444-1-Ig, 1:1000) were purchased from Proteintech (Rosemont, IL, USA).

Techniques: Activation Assay, Disruption, Inhibition