Characterization of the protein expression and localization of hnRNP family members during murine spermatogenesis / 亚洲男科学杂志(英文版)

Mammalian testis exhibits remarkably high transcriptome complexity, and spermatogenesis undergoes two periods of transcriptional cessation. These make the RNA-binding proteins (RBPs) the utmost importance during male germ cell development. Heterogeneous nuclear ribonucleoproteins (hnRNPs) are a large family of RBPs implicated in many steps of RNA processing; however, their roles in spermatogenesis are largely unknown. Here, we investigated the expression pattern of 12 hnRNP family members in mouse testes and found that most detected members are highly expressed in the testis. Furthermore, we found that most of the detected hnRNP proteins (hnRNPD, hnRNPK, hnRNPQ, hnRNPU, and hnRNPUL1) display the highest signals in the nuclei of pachytene spermatocytes, round spermatids, and Sertoli cells, whereas hnRNPE1 exclusively concentrates in the manchette of elongating spermatids. The expression of these hnRNP proteins showed both similarities and specificity, suggesting their diverse roles in spermatogenesis.

The role of human papillomavirus 16 early genes E2 and E6 and heterogeneous nuclear ribonucleoprotein E2 in cervical carcinogenesis and their interaction effect / 中华预防医学杂志

Objective@#To investigate the role of human papillomavirus (HPV) 16 early genes E2 and E6 and heterogeneous nuclear ribonucleoprotein (hnRNP) E2 and their interaction effects in the progression of the cervical cancer.@*Methods@#Women with normal cervix (NC), low cervical intraepithelial neoplasia (CIN Ⅰ) and high cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ) from the cervical lesions cohort in Jiexiu County of Shanxi Province from June 2014 to September 2014, and patients with cervical squamous cell carcinoma (SCC) treated at the Second Hospital of Shanxi Medical University in the same period were enrolled in this study. There were 257 participants, about 67 NC cases (26.07%), 69 CIN Ⅰ cases (26.85%), 68 CIN Ⅱ/Ⅲ cases (26.46%), and 53 SCC cases (20.62%), respectively. The information of demographic characteristics, life health habits and cervical lesions were collected by using the structured questionnaire. Cervical exfoliated cells and cervical biopsy tissues were collected to detect the infection of HPV16 and the protein expression levels of hnRNP E2, HPV16 E2 and E6. According to the median-value of the protein expression levels of hnRNP E2, HPV16 E2 and E6 and E2/E6 ratio in the NC group, the study participants were divided into the high and low expression groups/ratio groups. The multivariate logistic regression model was used to analyze the correlation between HPV16 early gene E2 and E6, hnRNP E2 and cervical cancer. The interaction effect was analyzed by using the generalized multifactor dimensionality reduction (GMDR) model.@*Results@#The ages of NC, CIN Ⅰ, CIN Ⅱ/Ⅲ and SCC groups were (47.00±9.07), (47.64±7.35), (46.37±8.67) and (51.26±8.03) years old, respectively. The multivariate logistic regression model analysis showed that the HPV16 E2 low expression, E6 high expression and E2/E6 low ratio could increase the risk of CIN Ⅱ/Ⅲ, about OR (95%CI) values 11.11 (1.63-75.56), 8.00 (1.28-50.04), and 9.75 (1.22-77.72), respectively and SCC, about OR (95%CI) values 14.22 (2.11-95.88), 10.33 (1.67-64.00), and 12.38 (1.56-97.91), respectively. The hnRNP E2 low expression could increase the risk of CIN Ⅱ/Ⅲ and SCC, about OR (95%CI) values 3.35 (1.39-8.10) and 5.53 (1.54-19.88). The result of GMDR showed that there were interaction effects of the hnRNP E2 low expression, HPV16 E2 low expression and HPV16 E6 high expression in both CIN Ⅱ/Ⅲ and SCC groups.@*Conclusion@#The HPV16 E2 low expression, HPV16 E6 high expression and hnRNP E2 low expression could increase the risk of high-grade cervical intraepithelial neoplasia and cervical cancer, and they might have an important interaction effect in the progression of the cervical cancer.

Effect of hnRNP K and its interaction with HPV16 on cervical intraepithelial neoplasia / 中华流行病学杂志

Objective To investigate the effect of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and its interaction with human papillomavims 16 (HPV16) on cervical intraepithelial neoplasia (CIN).Methods The participants included 67 women with normal cervix (NC),69 women with CIN Ⅰ and 68 women with CIN Ⅱ/Ⅲ in a community cohort of pathologically diagnosed women established in Jiexiu of Shanxi province,from June 2014 to June 2015.A structured questionnaire was used to collect the demographic data of the subjects and the related factors of cervical lesions.Cervical exfoliated cells and cervical tissues from biopsy or surgery were selected.The infection status of HPV16 was detected by flow-through hybridization.The protein expression levels of hnRNP K were evaluated by Western blot.SPSS 23.0 software was used to collate and analyze the data.To study the differences in demographic characteristics,related factors,hnRNP K protein and HPV16 infection among NC,CIN Ⅰ and CIN Ⅱ / Ⅲ groups,X2 test,trend x2 test,and Kruskal-Wallis H test were conducted.Multiple comparisons of hnRNP K protein in three groups were completed by using the Bonferroni method.The OR and its 95％CI of hnRNP K,HPV16 and CIN were calculated by using the unconditional logistic regression models.Two-way interactions between hnRNP K protein and HPV16 infection on CIN were analyzed by using additive model and related indicators.Results HPV16 infection rates were 10.4％ in women with normal cervix,14.5％ in women with CIN Ⅰ and 41.2％ in women with CIN Ⅱ/Ⅲ,respectively.The differences among three groups were significant (P＜0.001).Moreover,the infection rates of HPV16 gradually increased with the increasing severity of CIN (trend x2=18.512,P＜0.001).The differences in protein expression of hnRNP K among three groups were significant (H=48.138,P＜0.001) and the expressionincreased with the development of cervical lesionss (trend x2=21.765,P＜0.001).Results from the interaction analysis indicated that there were additive effects between high expression of hnRNP K protein and HPV16 in CIN Ⅱ/Ⅲ group compared with normal group (API=0.639,95％CI:0.083-1.196).In contrast,no such additive effect was found in CIN Ⅰ group.Conclusions HPV16 infection and over-expression of hnRNP K protein were associated with the increased risk of cervical intraepithelial neoplasia.There might be interaction between hnRNP K protein overexpression and HPV16 infection existed on the progress of CIN Ⅰ/Ⅲ.

Effect of hnRNP K and its interaction with HPV16 on cervical intraepithelial neoplasia / 中华流行病学杂志

Objective To investigate the effect of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and its interaction with human papillomavims 16 (HPV16) on cervical intraepithelial neoplasia (CIN).Methods The participants included 67 women with normal cervix (NC),69 women with CIN Ⅰ and 68 women with CIN Ⅱ/Ⅲ in a community cohort of pathologically diagnosed women established in Jiexiu of Shanxi province,from June 2014 to June 2015.A structured questionnaire was used to collect the demographic data of the subjects and the related factors of cervical lesions.Cervical exfoliated cells and cervical tissues from biopsy or surgery were selected.The infection status of HPV16 was detected by flow-through hybridization.The protein expression levels of hnRNP K were evaluated by Western blot.SPSS 23.0 software was used to collate and analyze the data.To study the differences in demographic characteristics,related factors,hnRNP K protein and HPV16 infection among NC,CIN Ⅰ and CIN Ⅱ / Ⅲ groups,X2 test,trend x2 test,and Kruskal-Wallis H test were conducted.Multiple comparisons of hnRNP K protein in three groups were completed by using the Bonferroni method.The OR and its 95％CI of hnRNP K,HPV16 and CIN were calculated by using the unconditional logistic regression models.Two-way interactions between hnRNP K protein and HPV16 infection on CIN were analyzed by using additive model and related indicators.Results HPV16 infection rates were 10.4％ in women with normal cervix,14.5％ in women with CIN Ⅰ and 41.2％ in women with CIN Ⅱ/Ⅲ,respectively.The differences among three groups were significant (P＜0.001).Moreover,the infection rates of HPV16 gradually increased with the increasing severity of CIN (trend x2=18.512,P＜0.001).The differences in protein expression of hnRNP K among three groups were significant (H=48.138,P＜0.001) and the expressionincreased with the development of cervical lesionss (trend x2=21.765,P＜0.001).Results from the interaction analysis indicated that there were additive effects between high expression of hnRNP K protein and HPV16 in CIN Ⅱ/Ⅲ group compared with normal group (API=0.639,95％CI:0.083-1.196).In contrast,no such additive effect was found in CIN Ⅰ group.Conclusions HPV16 infection and over-expression of hnRNP K protein were associated with the increased risk of cervical intraepithelial neoplasia.There might be interaction between hnRNP K protein overexpression and HPV16 infection existed on the progress of CIN Ⅰ/Ⅲ.

Progress of heterogeneous-nuclear ribonucleoproteins A1 in digestive system neoplasms / 肿瘤研究与临床

Heterogeneous-nuclear ribonucleoproteins A1 (hnRNP A1) is one of the important members of hnRNPs family. The function of hnRNP A1 is closely related to RNA transcription, mRNA translation, splicing of splicing sites, splicing site selection, pre-RNA maturation and degradation, cell proliferation and transformation. Overexpression of hnRNP A1 in various types of digestive system tumors is associated with poor prognosis and may serve as an early biomarker for cancer. This article reviews the recent advances in the development of hnRNP A1 and characterizes their roles in cellular and gene expression and in the development of human tumors, highlighting that hnRNP A1 is likely to be an important indicator for tumor prognosis and a potential drug target for the treatment.

Heterogeneous Nuclear Ribonucleoprotein A2B1 Exerts a Regulatory Role in Lipopolysaccharide-stimulated 38B9 B Cell Activation

Major histocompatibility complex (MHC) class II molecules, which are recognized for their primary function of presenting an antigen to the T cell receptor, are involved in various signaling pathways in B cell activation. We identified heterogeneous nuclear ribonucleoprotein (hnRNP) A2B1 as an MHC class II molecule-associated protein involved in MHC class II-mediated signal transduction in lipopolysaccharide (LPS)-stimulated 38B9 B cells. Although the function of hnRNP A2B1 in the nucleus is primarily known, the level of hnRNP A2B1 in the cytoplasm was increased in LPS-stimulated 38B9 cells, while it was not detected in the cytoplasm of non-treated 38B9 cells. The silencing of hnRNP A2B1 expression using siRNA disturbed B cell maturation by regulation of mitogen-activated protein kinase signaling, NF-κB activation, and protein kinase B activation. These results suggest that hnRNP A2B1 is associated with MHC class II molecules and is involved in B cell activation signaling pathways in LPS-stimulated 38B9 cells.

Cellular Localization Analysis of Enhanced Green Fluorescent Protein Tagged hnRNP A1 Under Stress / 天津医药

Objective To construct eukaryotic enhanced green fluorescent protein (EGFP) expressing recombinant plasmid, pEGFP-C1-hnRNP A1, which contains coding sequence of human hnRNP A1 (heterogeneous nuclear ribonucleo-protein A1), and to perform cellular localization analysis of EGFP tagged hnRNP A1 under stress. Methods Total RNA was isolated from HeLa cell used for synthesis of first-strand cDNAs using reverse primers that are specific for the 3′-un-translated region of hnRNP A1. hnRNP A1 gene fragments were then amplified by touch-down PCR from those cDNAs and inserted into pEGFP-C1 fluorescent bearing vector through EcoRⅠ/BamHⅡdouble enzyme digestion and T4 DNA Ligase connection. The recombinant pEGFP-C1-hnRNP A1 plasmid was transfected into HeLa cells and green fluorescent tagged fusion proteins was examined by Western blot and confocal fluorescence microscopy. Co-localization of EGFP-hnRNP A 1 with poly (A)+mRNA (the marker of the stress granules), or DCP1a (the marker of processome) were detected by RNA fluores-cence in situ hybridization and immunofluorescence. Results The pEGFP-C1-hnRNP A1 was sequenced and digested cor-rectly by restriction single/double enzyme. The green fluorescent fusion protein was also detected in transfected HeLa cell by Western blot and confocal fluorescence microscopy. EGFP-hnRNP A1 co-localizes with poly(A)+mRNA, but not DCP1a. Conclusion Recombinant eukaryotic plasmid of pEGFP-C1-hnRNP A1 was constructed successfully and expressed effec-tively. EGFP tagged hnRNP A1 takes part in forming stress granules.