AK4P1 is a cancer-promoting pseudogene in pancreatic adenocarcinoma cells whose transcripts can be transmitted by exosomes.

Adenylate kinase 4 pseudogene 1 (AK4P1) is a processed pseudogene whose function in cancer biology remains largely underexplored. Bioinformatics analysis suggested an association between the expression levels of adenylate kinase 4 (AK4) gene and AK4P1, as well as a clinical significance in relation to the increased transcription levels of AK4P1 in pancreatic adenocarcinoma (PAAD). In the present study, the expression levels of AK4P1 and AK4 were compared by RT-qPCR and western blotting between PAAD tissue and paired adjacent tissue. The level of AK4P1 transcript was compared between the circulating exosomes derived from patients with PAAD and those derived from healthy donors. Overall survival of the patients with PAAD with high or low expression levels of AK4P1 or AK4 was compared. AK4 gene expression level, in vitro cell viability and gemcitabine-induced apoptosis in PAAD cells with or without AK4P1 overexpression were also assessed using Cell Counting Kit-8 and TUNEL assays. It was identified that the transcription level of AK4P1 and the expression level of AK4 in PAAD tissue were significantly higher compared with those in paired non-cancerous tissue specimens. Transcription levels of AK4P1 and AK4 showed a significant relationship in PAAD. Circulating exosomes derived from patients with PAAD showed significantly higher level of AK4P1 transcript compared with that from circulating exosomes derived from blood samples of healthy donors. Patients with high expression of AK4P1 or AK4 exhibited significantly reduced overall survival compared with those with low expression. AK4P1 overexpression significantly upregulated the expression levels of AK4 in PAAD cells and rescued the viability and survival under gemcitabine challenge decreased by AK4 knockdown but not that by AK4 knockout. Treatment with exosomes secreted by AK4P1-overexpressing PAAD cells but not with those secreted by wild-type PAAD cells significantly increased the viability and survival under gemcitabine challenge of the recipient cells. These results suggested that AK4P1 affects the cellular biological functions of PAAD cells in vitro by upregulating the expression level of AK4. AK4P1 transcripts with elevated expression levels can be transmitted between PAAD cells through exosomes and exert pro-oncogenic effects in recipient cells.

Interleukin-17 receptor D is a favorable biomarker of glioblastoma.

BACKGORUND: Glioblastoma (GBM) is the most frequent glioma in adults. The prognosis of GBM is very poor and new prognostic biomarkers are in urgent need to better select high-risk patients and guide the individual treatments. METHODS: In our study, we compared the expression of Interleukin-17 receptor D (IL17RD) between GBMs and normal tissues from TCGA database, and detected IL17RD mRNA in 17 fresh GBM pairs with qPCR. With immunohistochemistry, we investigated the expression of IL17RD in 156 GBM tissues and further evaluated its clinical significance. The associations between IL17RD and clinicopathological factors were assessed by chi-square test. The prognostic significance of IL17RD was evaluated by univariate analysis with Kaplan-Meier method, and by multivariate analysis with Cox-regression Hazard model. RESULTS: The TPMs and mRNAs of IL17RD in GBM were substantially lower than those in normal brain tissues. The rates of low or high expression of IL17RD accounted for 41.67% and 58.33% respectively. IL17RD was significantly associated with higher survival rates of GBM. The 3-year overall survival rates of patients with low and high IL17RD were 7.2% and 19.5% respectively. In the Cox-regression model, the IL17RD expression was defined as an independent prognostic biomarker of GBM. Patients with high IL17RD expression had a more favorable outcome than those with low IL17RD. CONCLUSIONS: High IL17RD expression was an independent prognostic indicator of GBM, suggesting a more favorable prognosis. Our results suggested that IL17RD detection may help find the high-risk patients which may receive more severe surveillance and more individual treatments.

Regulation of microRNA miR-197-3p/CDC28 protein kinase regulatory subunit 1B (CKS1B) axis by Circular RNA hsa_circ_0000285 promotes glioma progression.

Circular RNA circ_0000285 is differentially expressed in several malignancies; however, its role in gliomas is under investigation. Reverse transcription quantitative polymerase chain reaction was conducted to evaluate the expression of circ_0000285, miR-197-3p, and CDC28 protein kinase regulatory subunit 1B (CKS1B) in glioma tissues and cells. Cell Counting Kit-8 and Transwell invasion assays coupled with Western blotting analysis using anti-Bax and anti-Bcl-2 antibodies were performed to evaluate cell proliferation, invasion, and apoptosis. Luciferase reporter and AGO2 RNA immunoprecipitation assays were conducted to verify the interaction between miR-197-3p and circ_0000285 or CKS1B. Xenograft tumor growth was evaluated in mice. We noted that circ_0000285 was highly expressed in glioma tissues and cells and that circ_0000285-silencing retarded tumor growth both in vitro and in vivo. This effect was mediated by the binding of circ_0000285 to miR-197-3p, which silenced CKS1B, an essential driver of glioma cell proliferation and invasion. Thus, circ_0000285 boosted glioma progression by regulating the miR-197-3p/CKS1B axis, highlighting a novel competing endogenous RNA circuit of glioma progression.

GNG12 Targeted by miR-876-5p Contributes to Glioma Progression Through the Activation of the PI3K/AKT Signaling Pathway.

Glioma is one of the most aggressive malignancies and has a poor survival rate. G protein subunit gamma 12 (GNG12), a member of G protein family, has been reported to participate in cancer disorders. However, the role and functional mechanism of GNG12 in glioma are not fully understood. The expression of GNG12 mRNA and miR-876-5p was measured by qRT-PCR. The level of GNG12 protein was measured by western blot. Cell proliferation and cell migration were monitored by CCK-8 assay and wound healing assay. The role of GNG12 on tumorigenicity in vivo was determined by animal models. The interaction between GNG12 and miR-876-5p was validated by dual-luciferase reporter experiment. The phosphorylation levels of PI3K and AKT were monitored by western blot. The upregulated expression of GNG12 was identified in tumor tissues and cells of glioma. GNG12 knockdown inhibited glioma cell growth and migration, and slowed tumor development in vivo. Besides, GNG12 knockdown weakened the phosphorylation levels of PI3K and AKT. GNG12 was verified to be a target of miR-876-5p whose enrichment suppressed the expression and function of GNG12. MiR-876-5p repressed glioma cell proliferation, migration, and the activity of PI3K/AKT signaling by targeting GNG12. MiR-876-5p-targeted GNG12 contributes to the malignant development of glioma by increasing the PI3K/AKT signaling activity.

A rare case of NIPT discrepancy caused by the placental mosaicism of three different karyotypes, 47,XXX, 47,XX,+21, and 48,XXX,+21.

BACKGROUND: Placental mosaicism is one of the major reasons for noninvasive prenatal testing (NIPT) discrepancy. Herein, we discovered a rare case of placenta with complex karyotypes that caused false-positive and false-negative results in noninvasive prenatal testing. METHODS: Next-generation sequencing (NGS) and Quantitative fluorescent polymerase chain reaction (QF-PCR) were performed on the cord blood sample, fetal tissues, and eight placental biopsies. Fluorescent In Situ Hybridization (FISH) and karyotyping were also carried to confirm the fetal genome status. RESULTS: The results suggested that the fetal chromosome was 47,XXX and the placenta had three karyotypes of 48,XXX,+21, 47,XX,+21, and 47,XXX. QF-PCR indicated that the extra chromosome 21 and chromosome X were all from the father. It is speculated that the zygote may have 48,XXX,+21 karyotype and trisomy rescue could be the main mechanism for the development of the homogeneous fetus and complex mosaic placenta. CONCLUSION: Overall, the complicated nature of our case underlines the importance of discussing with parents the possibility of both atypical and discordant results during preconfirmatory amniocentesis counseling and consent.

Negative interference in serum HBsAg ELISA from rheumatoid factors.

BACKGROUND: RF(Rheumatoid factor) is usually thought to cause positive interference in immunoassay. Recently, our study showed that high-concentration RFs caused negative interference as well as positive interference in serum HBsAg(Hepatitis B surface antigen) ELISA(Enzyme-linked immunosorbent assay), but it is unclear that RF causing negative interference is an anomaly produced by a certain ELISA kit or a common property of most of HBsAg ELISA kits. METHODS: Serum models were made by blending HBsAg-positive sera and high- or moderate-concentration RFs sera at the ratio of 1: 9, then one-step and two-step ELISA were adopted to determine HBsAg in serum models. RESULTS: No matter what kind of kit used, one-step ELISA showed that HBsAg S/CO( sample/cut off) values in serum models were significantly lower than original values. Bivariate correlations tests showed decline rates of HBsAg S/CO Values were not associated to serum RF concentrations ranging from 288 to 3560 IU/mL. HBsAg converted to be negative in 69.80% serum models with original-value ranging from 1.00 to 10.00, and in 2.68% serum models with higher original-value. RF causing decline of HBsAg S/CO value provided by one-step ELISA was more obvious than that provided by two-step ELISA. CONCLUSIONS: It is concluded that susceptibility of all HBsAg ELISA assays to interference from RF, leading to predominantly lower and in some cases "false-negative" results, and moreover, the lower the original HBsAg S/CO Value, the higher the false-negative rate.