Integrated proteome and physiological traits reveal interactive mechanisms of new leaf growth and storage protein degradation with mature leaves of evergreen citrus trees.

The growth of fruit trees depends on the nitrogen (N) remobilization in mature tissues and N acquisition from the soil. However, in evergreen mature citrus (Citrus reticulata Blanco) leaves, proteins with N storage functions and hub molecules involved in driving N remobilization remain largely unknown. Here, we combined proteome and physiological analyses to characterize the spatiotemporal mechanisms of growth of new leaves and storage protein degradation in mature leaves of citrus trees exposed to low-N and high-N fertilization in the field. Results show that the growth of new leaves is driven by remobilization of stored reserves, rather than N uptake by the roots. In this context, proline and arginine in mature leaves acted as N sources supporting the growth of new leaves in spring. Time-series analyses with gel electrophoresis and proteome analysis indicated that the mature autumn shoot leaves are probably the sites of storage protein synthesis, while the aspartic endopeptidase protein is related to the degradation of storage proteins in mature citrus leaves. Furthermore, bioinformatic analysis based on protein-protein interactions indicated that glutamate synthetase and ATP-citrate synthetase are hub proteins in N remobilization from mature citrus leaves. These results provide strong physiological data for seasonal optimization of N fertilizer application in citrus orchards.

Long non-coding RNA LINC00565 regulates ADAM19 expression through sponging microRNA-532-3p, thereby facilitating clear cell renal cell carcinoma progression.

Proven by publications, long non-coding RNAs (lncRNAs) play critical roles in the development of clear cell renal cell carcinoma (ccRCC). Although lncRNA LINC00565 has been implicated in the progression of various cancers, its biological effects on ccRCC remain unknown. This study aimed to investigate the biological functions of LINC00565, as well as its potential mechanism in ccRCC. Here, the expression data of mature microRNAs (miRNAs) (normal: 71, tumor: 545), messenger RNAs (mRNAs), and lncRNAs (normal: 72, tumor: 539) of ccRCC were acquired from The Cancer Genome Atlas (TCGA) database and subjected to differential expression analysis. Quantitative reverse transcriptase polymerase chain reaction analyzed the expression levels of LINC00565, miR-532-3p, and ADAM19 mRNA. TCGA database, dual-luciferase report detection, and Argonaute 2 RNA immunoprecipitation were utilized to confirm the relationships between LINC00565 and miR-532-3p and between miR-532-3p and ADAM19, respectively. The progression of ccRCC cells was determined via CCK-8, colony formation, scratch healing, and transwell assays. Western blot was applied to detect the protein levels of epithelial-mesenchymal transition markers and ADAM19. We herein suggested that LINC00565 was prominently upregulated in ccRCC tissues and cells. Knockdown of LINC00565 repressed cell progression. We further predicted and validated miR-532-3p as a target of LINC00565, and miR-532-3p could target ADAM19. Knockdown of LINC00565 resulted in ADAM19 level downregulation in ccRCC cells and suppressed miR-532-3p could restore ADAM19 level. Thus, the three RNAs constructed a ceRNA network. Overexpressed ADAM19 could eliminate the anticancer effects caused by knocking down LINC00565 on ccRCC cells. In conclusion, LINC00565 upregulated ADAM19 via absorbing miR-532-3p, thereby facilitating the progression of ccRCC cells.

Long Noncoding RNA MAGI2-AS3 Represses Cell Progression in Clear Cell Renal Cell Carcinoma by Modulating the miR-629-5p/PRDM16 Axis.

The objective of this study was to determine the regulatory mechanism of MAGI2-AS3 in clear cell renal cell carcinoma (ccRCC), thereby supplying a new insight for ccRCC treatment. Expression data in TCGA-KIRC were obtained. Target gene lncRNA for research was determined using expression analysis and clinical analysis. lncRNA's downstream regulatory miRNA and mRNA were predicted by bioinformatics databases. ccRCC cell malignant phenotypes were detected via CCK-8, colony formation, Transwell migration, and invasion assays. The targeting relationship between genes was assessed through dual-luciferase reporter gene analysis. Kaplan-Meier (K-M) analysis was carried out to verify the effect of MAGI2-AS3, miR-629-5p, and PRDM16 on the survival rate of ccRCC patients. MAGI2-AS3 expression in ccRCC tissue and cells was shown to be markedly decreased and its expression to continuously decline with tumor progression. MAGI2-AS3 suppresses ccRCC proliferation and migration. Dual-luciferase assay showed that MAGI2-AS3 binds miR-629-5p and that miR-629-5p binds PRDM16. In addition, functional experiments showed that MAGI2-AS3 facilitates PRDM16 expression by repressing miR-629-5p expression, thereby suppressing ccRCC cell aggression. K-M analysis showed that upregulation of either MAGI2-AS3 or PRDM16 significantly improves ccRCC patient survival, while upregulation of miR-629-5p has no significant impact. MAGI2-AS3 sponges miR-629-5p to modulate PRDM16 to mediate ccRCC development. Meanwhile, the MAGI2-AS3/miR-629-5p/PRDM16 axis, as a regulatory pathway of ccRCC progression, may be a possible therapeutic target and prognostic indicator of ccRCC.

Clinical efficacy of neoadjuvant chemohormonal therapy combined with laparoscopic radical prostatectomy in high-risk Prostate Cancer.

Objectives: To evaluate the clinical efficacy of neoadjuvant chemohormonal therapy (NCHT) combined with laparoscopic radical prostatectomy in high-risk prostate cancer (PCa). Methods: A randomized controlled trial was used in this study. Eighty patients with high-risk PCa treated in Tangshan Gongren Hospital from January 2017 to January 2019 were selected and randomly divided into two groups. The control group was given neoadjuvant endocrine therapy, while the research group was added NCHT to the control group. Three months later, the patients of two groups underwent laparoscopic radical prostatectomy. The changes of surgical indicators, adverse drug reactions, incidence of lower urinary tract symptoms, biochemical recurrence rate after follow-up, PSA progression-free survival and incidence of surgical complications were compared between the two groups. Results: After NCHT, the PSA level and prostate volume in the research group decreased significantly than those in the control group (P = 0.00). Surgical duration, postoperative hospital stay and retention time of drainage tube were significantly shorter and intraoperative blood loss was significantly less in the research group than those in the control group (P = 0.00). The incidence of lower urinary tract symptoms, biochemical recurrence and surgical complications in the research group were significantly lower than those in the control group, and the early recovery rate of urinary control and progression-free survival were significantly better than those in the control group (P < 0.05). Conclusion: NCHT combined with laparoscopic radical prostatectomy is a safe and effective treatment for high-risk PCa, which is worthy of promotion in clinical practice.

PRDM16, negatively regulated by miR-372-3p, suppresses cell proliferation and invasion in prostate cancer.

Prostate cancer (PCa) is one of the most prevalent malignant tumours. The alternation of microRNAs (miRNAs) expression is associated with prostate cancer progression, whereas its way to influence progression of prostate cancer remains elusive. The expression levels of PRDM16 mRNA and miR-372-3p in PCa cell lines were analysed using qRT-PCR. The protein expression of PRDM16 in PCa cell lines was also analysed using Western blot. CCK-8, wound healing and Transwell assays were applied to examine cell proliferation, migration, and invasion in prostate cancer cells, respectively. Dual-luciferase reporter assay was utilised to validate the interaction between miR-372-3p and PRDM16. In the present study, markedly decreased PRDM16 mRNA and protein expression levels were observed in prostate cancer cells. PRDM16 overexpression hampered cellular proliferation, migration, and invasion, while silencing PRDM16 had the opposite effect. Moreover, miR-372-3p could target the regulation expression of PRDM16. Rescue experiments demonstrated that upregulating miR-372-3p conspicuously restored the inhibitory effect of increased PRDM16 on cell proliferation, migration, and invasion in PCa. Overall, our study clarifies the biological role of miR-372-3p/PRDM16 axis in prostate cancer progression, which may be effective biomarkers for clinical treatment of prostate cancer.

miR-21-5p serves as a promoter in renal cell carcinoma progression through ARHGAP24 downregulation.

Renal cell carcinoma (RCC) is a highly recurrent aggressive tumor. This study works for the regulation of miR-21-5p on RCC cell functions and novel ideas for therapies of RCC. Isoform expression quantification data were offered by The Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma (TCGA-KIRC) to investigate differentially expressed miRNAs. The way miR-21-5p works on biological functions of RCC was examined with MTT and Transwell assays. The downstream targets of miR-21-5p were predicted using bioinformatics analysis. The binding of two researched objects was verified by the dual-luciferase method. TCGA data manifested a considerably high level of miR-21-5p in RCC tissue, while ARHGAP24 was significantly lowly expressed. miR-21-5p bound ARHGAP24 and stimulated RCC cell functions, whereas ARHGAP24 mimic could reverse such promotion. This work observed miR-21-5p, a stimulator in RCC, and it deteriorated this cancer via repressing its downstream target gene ARHGAP24 expression.

MicroRNA-155-5p Targets NR3C2 to Promote Malignant Progression of Clear Cell Renal Cell Carcinoma.

BACKGROUND: The molecular heterogeneity of clear cell renal cell carcinoma (ccRCC) leads to a high mortality of the disease, which seriously threatens the life of patients. Therefore, this study explored the functional significance and mechanism of microRNA-155-5p and nuclear receptor subfamily 3 group C member 2 (NR3C2) in the regulation of ccRCC. METHODS: Expression levels of microRNA-155-5p and NR3C2 mRNA in ccRCC cells were analyzed by qRT-PCR, and the protein expression of NR3C2 in human ccRCC cells was measured by Western blot. Biological functions were determined through a series of in vitro experiments. The interaction between microRNA-155-5p and NR3C2 was tested by luciferase reporter gene assay. In addition, the effect of overexpressed or silenced microRNA-155-5p on cell phenotypes was evaluated in ccRCC cells. RESULTS: Experimental data suggested that overexpression or silencing of microRNA-155-5p in ccRCC could boost or suppress cancer cell proliferation and other malignant behaviors. Rescue experiments revealed that microRNA-155-5p facilitated the proliferation, migration, and invasion and suppressed the apoptosis of ccRCC by directly inhibiting the expression of NR3C2. CONCLUSIONS: This is the first study to generate new insights into the role of microRNA-155-5p/NR3C2 interaction in promoting the process of ccRCC, and it is possible to bring a turning point for the treatment of ccRCC.

Inhibition of microRNA-300 inhibits cell adhesion, migration, and invasion of prostate cancer cells by promoting the expression of DAB1.

Prostate cancer (PC) is the most common malignancy in men. As per recent findings, microRNA-300 (miR-300) were found to be overexpressed in numerous types of cancers. In this study, we aimed to explore the effects of miR-300 on the adhesion, invasion, and migration of PC cells by targeting Disabled 1 (DAB1). Firstly, the regulatory role of miRNAs on DAB1 was predicted by screening PC-related differentially expressed genes (DEGs). Immunohistochemistry was applied to determine the positive protein expression of DAB1, after which the target relationship between miR-300 and DAB1 was examined. Loss-of-function and gain-of-function experiments were conducted to determine cell proliferation, adhesion, migration, invasion capability, and cell cycle of PC cells. Our data illustrated that DAB1 had a low expression, while miR-300 was expressed at a relatively high level in PC tissues. Moreover, our clinicopathological analysis revealed that there was a correlation between miR-300 and tumor, node, metastases stage, Gleason score, and lymph node metastasis of PC patients. DAB1 was also found to be poorly expressed in PC based on the findings from the microarray analysis. The results from dual-luciferase reporter gene assay corroborated that miR-300 interacts with DAB1. Importantly, overexpression of miR-300 and/or si-DAB1 resulted in the enhancement of RAC1, MMP2, MMP9, CyclinD1, and CyclinE expressions, whereas the expression of DAB1 and Rap was reduced in PC cells, thus suggesting that down-regulated miR-300 suppressed proliferation, adhesion, migration, and invasion of PC cells. Collectively, our results provided evidence that down-regulation of miR-300 inhibits the adhesion, migration, and invasion of PC cells.

Systematic analysis on the GSTM1 null phenotype and prostate cancer risk in Chinese people.

OBJECTIVE: Glutathione S-transferase M 1 (GSTM1) is implicated as a risk factor for prostate cancer. However, this issue is not clear in Chinese population. This systemic analysis was conducted to evaluate the effect of GSTM1 null genotypes on prostate cancer risk in Chinese. METHODS: Published studies investigating the associations between GSTM1 null genotypes and the risk of prostate cancer in China were identified by using a predefined search strategy. Main statisticals were pooled and estimated according to the primarily reported data. RESULTS: The prevalence of the GSTM1 null genotype was higher in prostate cancer patients than in controls, with significance. CONCLUSION: The GSTM1 null genotypes is associated with increased risk of prostate cancer in Chinese.

[Inhibitory effect of dendritic cells induced activated cytotoxicity T lymphocyte combined with MAGE-1 nonapeptide on transplanted human hepatocyte carcinoma in nude mice].

OBJECTIVE: To observe the inhibitory effect of dendritic cells (DCs) activated cytotoxicity T lymphocyte (CTL) combined with MAGE-1 nonapeptide on transplanted human hepatocyte carcinoma (HCC) in nude mice. METHODS: A model of HCC transplanted tumor was established by injecting BEL-7402 cell line HCC cells subcutaneously on the back of nude mice. Successful transplantation rate was 73%. Specific CTLs (1 x 10(6)), which were activated by DCs combined with MAGE-1 nonapeptide, were injected into the site of transplanted tumor (group A, n = 5). Another group of 17 mice were treated with same amounts of different kinds of cells, and they were divided into groups B, C, D, E, and F. The growth of tumor was observed, and pathological examination was also done. RESULTS: (1) The activated lymphocytes induced by DCs combined with MAGE-1 nonapeptide could suppress the growth of tumor and reduce the tumor size. In group A, 5/5 mice survived for at least two weeks, while the tumors grew rapidly and the majority of the mice died within two weeks in other groups (groups B, C, D, E, F) (P < 0.01). (2) Extensive necrosis and apoptosis were found in transplanted tumors in group A. CONCLUSIONS: The DCs combined with MAGE-1 nonapeptide could not only inhibit the growth of HCC, but also result in produce death and apoptosis of HCC, hence preventing tumor metastasis and recurrence. The mechanism underlying tumor immunization resulted from DCs might be enhanced in apoptosis of tumor cells. MAGE-1 nonapeptide combined with DCs might be a potential novel tumor vaccine for the treatment of HCC.