Differential Gene Expression Profiles of PPP2R5C-siRNA-Treated Malignant T Cells

Abstract

Recently, alterations in the expression pattern of PPP2R5C associated with malignant transformation have been characterized, and PPP2R5C overexpression was demonstrated in leukemias. To confirm the role of PPP2R5C in proliferation and its molecular mechanism, three PPP2R5C-siRNAs and a scrambled nonsilencing siRNA control were used to treat Molt-4 and Jurkat T cells. After nucleofection, PPP2R5C expression and biological consequences based on a highly efficient and specific PPP2R5C-siRNA were demonstrated by qRT-PCR, CCK-8 assay, Annexin V/PI, and flow cytometry. The global gene expression profile of PPP2R5C-siRNA-treated Jurkat T cells was established. A significant reduction in the PPP2R5C mRNA level was observed at 24 to 72 h in Molt-4 and Jurkat T cells with all of the PPP2R5C-siRNAs. The proliferation rate of Molt-4 and Jurkat T cells transfected with different PPP2R5C-siRNAs was significantly decreased at 72 h compared with the control ( p < 0.05). However, the transfected cells did not show a significant increase in Annexin V/PI-positive cells (apoptosis). The highly efficient PPP2R5C-siRNA2 was used to treat Jurkat T cells for gene expression profile analysis. In total, 439 genes were upregulated, and 524 genes were downregulated at least twofold in PPP2R5C-siRNA-treated Jurkat T cells. Changes in signaling pathway genes closely related to the TCR, Wnt, calcium, MAPK, and p53 signaling pathways were observed. In conclusion, the suppression of PPP2R5C by RNA interference could effectively inhibit the proliferation of leukemic T cells, the PPP2R5C-siRNA treatment altered gene expression profiles, and the differential expression of the glycogen synthase kinase 3 beta (GSK-3b), ataxia telangiectasia mutated (ATM), and Mdm2 p53 binding protein homolog (MDM2) genes may play an important role in the effects of PPP2R5C knockdown in Jurkat T cells.

Characteristics of TCRζ, ZAP-70, and FcɛRIγ Gene Expression in Patients with T- and NK/T-Cell Lymphoma

“…PCR was performed in a 20 mL volume with *1 mL of cDNA, 0.5 mM of each primer pair, 9 mL of 2.5 · Real Master Mix (Tiangen Biotech, Beijing, China), and 9 mL of dH 2 O. The primer details are listed in Table 3 and the sequences and PCR conditions have been previously described (Huang et al, 2012;Li et al, 2012b;Chen et al, 2013). The relative amount of the genes of interest and the b2M reference gene was measured in two independent assays.…”

Loss of PPARγ in endothelial cells leads to impaired angiogenesis

“…Furthermore, RNA sequencing analysis suggests a significant attenuation of CCND1 expression in PPARγ-silenced PMVECs (Table S1B). CCND2 is commonly upregulated upon Wnt activation associated with proliferation, (Yasui et al, 2006), whereas downregulation of PPP2R5C inhibits cell proliferation (Chen et al, 2013). Moreover, Fig.…”

Evaluation of PPP2R5C gene expression in Iranian patients with B-Acute lymphoblastic leukemia and its association with clinical and laboratory findings

“…Two different mutations in lung cancer and intestinal cancer can disrupt B56γ -P53 interaction (12,20). It has been showed that PPP2R5C suppression by RNAi can inhibit the proliferation of the Molt4 and Jourkat T cells (21). It also has been showed that the proliferation of the PPP2R5C-siRNAtreated CML cell line was decreased in K562 Cell line (22).…”