Global deletion of the RNA decapping enzyme Dcp2 postnatally in male mice results in infertility.

The post-transcriptional regulation of gene expression plays an important role in many essential biological processes. The RNA decapping enzyme Dcp2 is a crucial enzyme involved in RNA degradation. Dcp2 proteins are highly expressed in the testis and brain in adult mice. This study aimed to investigate the in vivo functions of Dcp2. An inducible Dcp2 knockout mouse model was established. No obvious health abnormalities were observed after postnatal global deletion of Dcp2 in male mice. However, Dcp2-deleted male mice were infertile and showed Sertoli cell vacuolization and germ cell degeneration. Dcp2 deletion resulted in testicular atrophy, reduced number of epididymal sperm, and increased apoptosis in seminiferous tubules. However, spermatocyte-specific deletion of Dcp2 did not compromise the fertility. The findings of this study indicated that Dcp2 was important for spermatogenesis and male fertility.

MicroRNA-141-3p/200a-3p target and may be involved in post-transcriptional repression of RNA decapping enzyme Dcp2 during renal development.

The RNA decapping enzyme Dcp2 is a crucial enzyme involved in the process of RNA turnover, which can post-transcriptionally regulate gene expression. Dcp2 has been found to be highly expressed in embryonic, but not adult, kidneys. Here we showed that Dcp2 mRNA was expressed, but Dcp2 proteins were absent, in mouse kidneys after postnatal day 10 (P10). In kidneys of adult Dcp2-IRES-EGFP knock-in mice, Dcp2 was undetectable but EGFP was expressed, indicating that Dcp2 mRNA was not completely silenced in adult kidneys. Using luciferase reporter assays, we found that miR-141-3p/200a-3p directly targeted the 3' UTR of Dcp2 mRNA. Overexpression of miR-141-3p and miR-200a-3p downregulated endogenous Dcp2 protein expression. Furthermore, miR-141-3p and miR-200a-3p expression was low in embryonic kidneys but increased dramatically after P10 and was negatively correlated with Dcp2 protein expression during renal development. These results suggest miR-141-3p/200a-3p may be involved in post-transcriptional repression of Dcp2 expression during renal development. ABBREVIATIONS: IRES: internal ribosome entry site; EGFP: enhanced green fluorescent protein; UTR: untranslated region.

[Analysis of microchimerism in living renal transplantation between mothers and children relationship].

OBJECTIVE: To compare the microchimerismic and rejection rates in living donor kidney transplant recipients in mother and child relations and other relations. METHODS: This retrospective single-center study enrolled 130 recipients to receive allografts from living related donors from 2004 to 2008 at our hospital. They were followed up for 1 - 5 years. The demographic data of the study population were analyzed by basic statistical methods. A total of 43 recipient blood samples were collected for the detection of microchimerism by the assays of short tandem repeat (STR) and sex-determining region-y gene (SRY) polymerase chain reaction (PCR). RESULTS: The 1-year patient/graft survival rates were 93.8% and 92.3% respectively. And there was no significant differences between mother and child group and other relative group. Forty-six biopsy samples were collected from 46 recipients. Twenty-six (20.0%) cases had the occurrences of acute rejection episodes in different Banff degrees as proven by biopsy. 53.8% (14/26) cases were mother and child renal transplantation, higher than other relative (46.2%, 12/26). The mother donor kidney transplant recipients had about a twice higher rejection rate (30.4% vs 14.3%, P = 0.028) and a twice higher microchimerismic rate (25.0% vs 14.8%) than other relative. CONCLUSION: Compared with other relations, the mother donor kidney recipients tend to have higher rates of microchimerism and acute rejection. And the special immune effect in mothers and children renal transplantation may influence its outcomes.

[Preliminary exploration in proteomic-based mechanistic analysis of patients with acute renal allograft rejection].

OBJECTIVE: To explore the application of proteomics in the mechanistic analysis of acute rejection (AR). METHODS: Quantified proteomics with isobaric tags for relative and absolute quantitation (iTRAQ) labeling was utilized to identify the protein profiling between the transplantation patients with (n = 5) or without AR (n = 8) from 2008 to 2010. RESULTS: Among the 179 identified proteins, 66 proteins in AR patients had at least a 2-fold change as compared with those without AR. The results demonstrated the dominant processes and responses associated with inflammation and complement activation. It was consistent with the underlying immune rejection associated with AR. Moreover, the results also indicated that high-coagulation state existed in AR patients. A number of transcription factors were identified in AR patients, including nuclear factor-κB, signal transducer and activator of transcription 1, signal transducer and activator of transcription 3. The analysis of transcription regulation networks suggested that the cross-talks among these key transcription factors might play an important role in the acute response and activation of coagulation system. CONCLUSION: The application of proteomics provides a new strategy of mechanistic analysis in AR.

miRNA-1297 induces cell proliferation by targeting phosphatase and tensin homolog in testicular germ cell tumor cells.

To investigate the role of miR-1297 and the tumor suppressor gene PTEN in cell proliferation of testicular germ cell tumors (TGCT). MTT assays were used to test the effect of miR-1297 on proliferation of the NCCIT testicular germ cell tumor cell line. In NCCIT cells, the expression of PTEN was assessed by Western blotting further. In order to confirm target association between miR-1297 and 3'-UTR of PTEN, a luciferase reporter activity assay was employed. Moreover, roles of PTEN in proliferation of NCCIT cells were evaluated by transfection of PTEN siRNA. Proliferation of NCCIT cells was promoted by miR-1297 in a concentration-dependent manner. In addition, miR-1297 could bind to the 3'-UTR of PTEN based on luciferase reporter activity assay, and reduced expression of PTEN at protein level was found. Proliferation of NCCIT cells was significantly enhanced after knockdown of PTEN by siRNA. miR-1297 as a potential oncogene could induce cell proliferation by targeting PTEN in NCCIT cells.