Fig. 7: Knockdown of LFPRLR in malignant human B cells reduces BCL2 and MYC expression. | Communications Biology

Fig. 7: Knockdown of LFPRLR in malignant human B cells reduces BCL2 and MYC expression.

From: Isoform-specific knockdown of long and intermediate prolactin receptors interferes with evolution of B-cell neoplasms

Fig. 7

a PRL secretion level by ELISA and b viable cell number after treatment with rabbit anti-PRL or rabbit normal serum, assessed by MTS assay in malignant human B-cell lines. P-values by unpaired t test at different concentrations of anti-PRL or normal serum in b: SU-DHL-6 (P = 0.0009 in 1:1600 and P < 0.0001 in 1:800, 1:400, 1:200, 1:100 and 1:50), Karpas-422 (P = 0.0283 in 1:1600, P = 0.0016 in 1:800, P = 0.0004 in 1:200 and P < 0.0001 in 1:400, 1:100 and 1:50), VAL (P = 0.0066 in 1:1600, P = 0.0178 in 1:100 and P = 0.0002 in 1:50) and OCI-LY18 (P = 0.0253 in 1:1600, P = 0.0007 in 1:800, P = 0.0051 in 1:100, P = 0.0007 in 1:50). P-values were calculated by unpaired t test (±SD). P values for 1:50 are indicated in the figure. ns non-significant. c, d LFPRLR mRNA levels and expression of MYC, BCL2 and PRL transcripts in malignant human B-cell lines (Karpas-422, SU-DHL-6, VAL and OCI-LY18) treated with 73 nM of control SMO or LFPRLR SMO for 48 h. Each qPCR sample was run in 3 technical replicates. One representative of 3 independent experiments is shown. UBB was used as the house keeping gene in qPCR. P-values are calculated by unpaired t test from 3 technical replicates (±SEM). Cell lines highly sensitive to low concentration (73 nM) of LFPRLR SMO are shown in (c) and cell lines less sensitive to LFPRLR SMO at 73 nM are shown in d. e, f Immunoblotting to compare the expression of MYC and BCL2 proteins in human DLBCL cell lines (Karpas-422, SU-DHL-6, VAL, and OCI-LY18) treated with 73 nM of control SMO or LFPRLR SMO for 48 h. One representative of 3 independent experiments is shown. β-actin was used as the loading control in immunoblotting. Cell lines highly sensitive to low concentration (73 nM) of LFPRLR SMO are shown in e and cell lines less sensitive to LFPRLR SMO at 73 nM are shown in f. g Signaling of PRL through the LF/IFPRLR induces the expression of MYC and BCL2 in overt human DLBCL and B-ALL, thereby promoting malignant B-cell growth and survival. LFPRLR SMO thus represents a promising candidate to treat these malignancies. g was created with BioRender.com.

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