LINC00467 induces melanoma deterioration by targeting miR-485-5p/p21 activated kinase 1.

Background: The purpose of the current research was to investigate the biological roles of LINC00467 in inducing melanoma deterioration. Methods: Differential level of LINC00467 in melanoma tissues and its prognostic value were analyzed in GEPIA, which were further confirmed in clinical samples we collected. Regulatory effects of LINC00467 on proliferation, migration and invasion capacities of A375 and SKMEL1 cell lines were examined by a series of functional experiments. Potential downstream targets of LINC00467 were identified through dual-luciferase reporter assay, and their synergistic role in melanoma process was finally explored by rescue experiments. Results: LINC00467 was up-regulated in melanoma samples, but it did not have a prognostic potential in melanoma. LINC00467 has the capacities to stimulate proliferation, migration and invasion of A375 and SKMEL1 cell lines. The feedback loop LINC00467/miR-485-5p/PAK1 was identified, which was responsible for inducing melanoma deterioration. Conclusions: LINC00467 stimulates proliferation, migration and invasion capacities of melanoma via targeting miR-485-5p to upregulate PAK1, which provides potential targets for treatment of melanoma.

Ring finger 220 promotes the stemness and progression of colon cancer cells via Ubiquitin specific peptidase 22-BMI1 axis.

Colorectal cancer (CRC) is ranked as the third most common malignancy worldwide. Therefore, it is urgent to screen novel and effective molecular drug targets for colorectal cancer therapeutics. In this study, the specific role and related mechanism underlying Ring finger (RNF) 220 in colon cancer were investigated. Firstly, RT-PCR assay was used to compare differences between expression levels of RNF220 in colorectal tumor and normal tissues. Western blot and RT-PCR assays were applied to examine the protein levels of RNF220 in normal colonic mucosa and colorectal cancer cells. We found that RNF220 was upregulated in colorectal cancer in patients and cell models. RNF220 promoted the proliferation and migration, invasion of colorectal cancer cells through BrdU incorporation, clone formation, transwell and wound healing assays. Spheroid formation and western blot assays illustrated that RNF220 promoted the stemness of colorectal cancer cells. Moreover, we found that RNF220 regulated BMI1 expression through USP22 by western blot. Finally, we discovered that RNF220 facilitated tumor growth in vivo through establishment of subcutaneous xenograft tumor mice model. In conclusion, RNF220 promoted the stemness and progression of colon cancer cells via the USP22-BMI1 axis.

Prognostic and clinicopathological significance of pretreatment mean platelet volume in cancer: a meta-analysis.

OBJECTIVE: Our study aimed to evaluate the prognostic and clinicopathological significance of pretreatment mean platelet volume (MPV) on cancer by using meta-analysis of published studies. DESIGN: Meta-analysis. DATA SOURCES: Relevant studies available before 22 December 2019 were identified by searching MEDLINE, EMBASE. ELIGIBILITY CRITERIA: All published studies that assessed the prognostic and clinicopathological significance of pretreatment MPV on cancer were included. DATA EXTRACTION AND SYNTHESIS: Studies were identified and extracted by two reviewers independently. The HR/OR and its 95% CIs of survival outcomes and clinicopathological parameters were calculated. RESULTS: A total of 38 eligible studies (41 subsets) with 9894 patients with cancer were included in the final meta-analysis. MPV level was not significantly associated with both overall survival (HR 0.98, 95% CI 0.84 to 1.14) and disease-free survival (HR 1.22, 95% CI 0.86 to 1.73) of patients with cancer. Neither advanced nor mixed-stage tumour patients showed significant association between MPV and overall survival (HR 1.36, 95% CI 0.96 to 1.94, HR 0.90, 95% CI 0.74 to 1.09). However, high MPV had the strongest relationship with poor overall survival (HR 2.01; 95% CI 1.08 to 3.41) in gastric cancer, followed by pancreatic cancer (HR 1.54; 95% CI 1.31 to 1.82). Whereas in the subgroup using receiver operating characteristic curve method to define cut-off values, low MPV was significantly related to poor overall survival (HR 0.78, 95% CI 0.64 to 0.95). In addition, MPV had no significant association with age (OR 0.96, 95% CI 0.90 to 1.02), sex (OR 1.04, 95% CI 1.00 to 1.09), depth of cancer invasion (OR 0.90, 95% CI 0.77 to 1.04) and tumour stage (OR 0.91, 95% CI 0.78 to 1.07). CONCLUSIONS: Pretreatment MPV level is of no clearly prognostic significance in cancers and no significant association with clinicopathological parameters of patients with cancers.

Erythroblast transformation-specific 2 correlates with vascular smooth muscle cell apoptosis in rat heterotopic heart transplantation model.

BACKGROUND: Cardiac allograft vasculopathy (CAV) decreases the long-term survival of heart transplantation recipients. Vascular smooth muscle cell (VSMC) apoptosis is an important pathological feature of CAV. Erythroblast transformation-specific 2 (Ets-2), as a transcription factor, participates in cell apoptosis and plays an important role in organ transplantation. METHODS: Hearts from Wistar-Furth (WF:RT1u) rats were heterotopically transplanted into Lewis (Lew:RT1(l)) rats without immunosuppression. Additional syngeneic heterotopic cardiac transplantations were performed in Lewis rats. HE staining was used to identify CAV. Ets-2 expression was examined by western blot. Ets-2 tissue location was examined by immunohistochemical assay and double immunostaining. Cleaved caspase 3 expression was detected by western blot. Co-localization of Ets-2 and cleaved caspase 3 was detected by double immunostaining. Ets-2, p53, cleaved caspase 3 and Bcl-xl expression in rat VSMC line A7R5 was examined after Ets-2 siRNA transfection. TUNEL assay was applied to detect A7R5 apoptosis with or without ETS-2 siRNA transfection. Immunoprecipitation was performed to explore the interaction between Ets-2 and p53. RESULTS: Ets-2 expression decreased in the allograft group but had no obvious change in the isograft group. Meanwhile, the phenomenon of CAV was observed in the allograft group and there is neointima formation in the isograft group which is not obvious compared with allograft group. Additionally, Ets-2 expression was opposite to VSMC apoptosis in the allograft group. In vitro, Ets-2 siRNA transfection in A7R5cells resulted in enhanced cell apoptosis. Finally, Ets-2 interacted with p53. CONCLUSIONS: Ets-2 might inhibit VSMC apoptosis via p53 pathway. The results further elucidate the molecular mechanism of VSMC apoptosis after heart transplantation during CAV and provide theoretical basis for seeking new specific drug targets for CAV prevention and treatment.

Overcoming the diffraction limit of imaging nanoplasmonic arrays by microspheres and microfibers.

Super-resolution microscopy by microspheres emerged as a simple and broadband imaging technique; however, the mechanisms of imaging are debated in the literature. Furthermore, the resolution values were estimated based on semi-quantitative criteria. The primary goals of this work are threefold: i) to quantify the spatial resolution provided by this method, ii) to compare the resolution of nanoplasmonic structures formed by different metals, and iii) to understand the imaging provided by microfibers. To this end, arrays of Au and Al nanoplasmonic dimers with very similar geometry were imaged using confocal laser scanning microscopy at λ = 405 nm through high-index (n~1.9-2.2) liquid-immersed BaTiO3 microspheres and through etched silica microfibers. We developed a treatment of super-resolved images in label-free microscopy based on using point-spread functions with subdiffraction-limited widths. It is applicable to objects with arbitrary shapes and can be viewed as an integral form of the super-resolution quantification widely accepted in fluorescent microscopy. In the case of imaging through microspheres, the resolution ~λ/6-λ/7 is demonstrated for Au and Al nanoplasmonic arrays. In the case of imaging through microfibers, the resolution ~λ/6 with magnification M~2.1 is demonstrated in the direction perpendicular to the fiber with hundreds of times larger field-of-view in comparison to microspheres.

Pyruvate kinase isoenzyme M2 expression correlates with survival of cardiomyocytes after allogeneic rat heterotopic heart transplantation.

The aim of our study was to assess correlations between PKM2 and the survival of cardiomyocytes after heart transplantation in rat. The PKM2, Bcl-xl, active caspase-3 proteins were detected by western blot, and PKM2 was testified by immunohistochemistry and immunofluorescence. At the same time, active caspase-3, α-actinin, VCAM-1, and CD4 were detected by immunofluorescence. Compared with rare expression in syngeneic Lewis rat hearts, the PKM2 protein level in allogeneic hearts was detected at various survival times after transplantation, which prominently expressed on day five postoperatively. In addition, we examined the expression of Bcl-xl and active caspase-3 in allogeneic hearts, which has a similar expression pattern with PKM2. Immunohistochemical and immunofluorescent methods displayed that PKM2 was widely expressed in cardiac tissue, and active caspase-3 was also expressed in cardiomyocytes. However, the PKM2 was not expressed in T cells and other immune response cells. These results suggested that PKM2 may regulate the survival of cardiomyocytes in acute rejection after heart transplantation in rat.

Expression of HSPA12B in acute cardiac allograft rejection in rats.

HSP70 may play a more important role in regulating antigen-specific immune response than other HSPs; however, HSPA12B production in transplanted heart remains obscure, which was identified as the newest member of the HSP70 family. In the current study, we performed a heart transplantation model in adult rats and investigated the dynamic changes of HSPA12B expression in the cardiac grafts. The cardiac grafts of allogeneic (Wistar-Lewis rat) and syngeneic (Lewis-Lewis rat) rat models were subjected to histopathological and immunohistochemical analyses for HSPA12B expression on days 0-7 after operation. We also examined the expression profiles of active caspase-3, whose changes were correlated with the expression profiles of HSPA12B. Our results demonstrated that HSPA12B protein exhibited biphasic patterns in transplanted heart. The first expression phase correlated with ischemical reperfusion injury over 2 days post-transplant. The second peak of HSPA12B expression was found only in allografts on day 5, concurrent with the expression of caspase-3. Immunohistochemical assay showed that compared with rare expression in isografts, there were significant protein expressions of HSPA12B and caspase-3 in heart allografts from day 5 to 7 post-transplant. Furthermore, double immunofluorescence staining for active caspase-3 and HSPA12B in isografts and allografts at day 5 post-transplant were analyzed and colocalization of HSPA12B/active caspase-3 was detected in allografts. In conclusion, this is the first description of HSPA12B expression in acute cardiac allograft rejection. Our results suggested that HSPA12B might play crucial roles in heart pathophysiology after transplantation.

Correlation of serum- and glucocorticoid-regulated kinase 1 expression with ischemia-reperfusion injury after heart transplantation.

IRI of a transplanted heart may result in serious early and late disadvantageous effects such as increased allograft immunogenicity, primary graft dysfunction, and initiation of fibroproliferative cascades that compromise the survival of the recipient. Sgk-1 has recently been linked to cell growth and survival. It has been reported that through a renal transplantation model, Dexa increases Sgk-1 expression and therefore protects from renal IRI. In our current study, we aim to assess the expression of Sgk-1 and its protective effects on cardiomyocyte IRI after heart transplantation. Heart allograft model was performed from Wistar into Lewis, and isograft model was from Lewis into Lewis. Grafts were then harvested at one, six, 12, or 24 h post-transplantation for Sgk-1 expression analyses. In some groups, part donors were treated with Dexa 2 h prior at doses of 0.05, 0.5 and 2 mg/BWkg, respectively. Sgk-1 expression was markedly increased in grafted heart 6-12 h post-transplantation in both the allogenic and isogenic models. Immunostaining experiments confirmed that Sgk-1 was expressed in cardiomyocytes rather than infiltrated immune cells. Furthermore, Dexa treatment significantly increased Sgk-1 expression and the donor cardiomyocyte injury was greatly minimized by Dexa treatment. These results suggest that induction of Sgk-1 might explain some of the beneficial impact of corticosteroids in IRI and hence might have therapeutic implications.

Upregulation of α-enolase in acute rejection of cardiac transplant in rat model: implications for the secretion of interleukin-17.

Acute allograft rejection remains a major problem in solid organ transplantation. The enzyme α-enolase has been shown to induce an immune response in cardiac transplantation. In this study, we investigated the role of α-enolase in acute allograft rejection in a rat model of heart transplantation. Hearts from either (WF: RT1(u) ) or (Lew: RT1(1) ) rats were transplanted into (Lew: RT1(1) ) rats. No rejection occurred in the isograft group, for which the median survival time was >168 days, whereas the median survival time of the allograft group was significantly less at 10 ± 2.1 days (n = 8 per group, p < 0.001). Increased inflammation was observed in allografts, including increased α-enolase expression and increased numbers of infiltrating CD4(+) T cells (p < 0.05). By immunohistochemical staining, we confirmed that α-enolase was expressed not only in myocardial cells but also in the infiltrating lymphocytes. However, on the fifth day after transplantation, α-enolase expression was no longer observed in the lymphocytes (n = 3, p < 0.001). In contrast, no lymphocytes were found in isografts after transplantation (n = 3, p < 0.001). α-enolase expression was increased in lymphocytes, which are implicated in the acute rejection of cardiac transplants. Intragraft α-enolase inhibition may be useful as an adjuvant therapy to systemic immunosuppression in heart transplantation.

Polycomb group oncogene RING1 is over-expressed in non-small cell lung cancer.

Ring finger protein 1 (RING1) have recently been reported to be related to aggressive tumor features in Prostate Cancer and urothelial carcinoma of the bladder. However, the role of RING1 in non-small-cell lung cancer (NSCLC) tumorigenesis has never been elucidated. So we aimed at investigating the potential role of RING1 in NSCLC. RING1 expression was evaluated by Immunoblot in 8 paired fresh lung cancer tissues and immunohistochemistry on 69 paraffin-embedded sections from 2006 to 2009. Furthermore, flow-cytometry and RNA interference were performed to analyse the role of RING1 in A549 cells. We showed that the expression level of RING1 was significant increased in lung cancer as compared with the adjacent normal tissue. High expression level of RING1 was associated with TNM stage (P = 0.013), and RING1 was positively related with proliferation marker Ki67 (P < 0.05). Moreover, RING1 knockdown induces growth suppression of human lung cancer cells through G1/S cell cycle phase arrest in vitro. Kaplan-Meier survival curves showed that high expression level of RING1 was associated with poor prognosis (P = 0.03). On the basis of these results, we suggested that RING1 protein expression may be a favorable independent prognostic parameter for non-small cell lung cancer.

Fiber coupling to BaTiO3 glass microspheres in an aqueous environment.

Compact microspheres with high-quality (Q) whispering gallery modes are required for many applications involving liquid immersion, such as sensing nanoparticles and studying resonant radiative pressure effects. We show that high-index (1.9 and 2.1) barium titanate glass (BTG) microspheres are perfect candidates for these applications due to their high-Q (∼10(4) in the 1100-1600 nm range) resonances evanescently excited in spheres with diameters of 4-15 µm. By reattaching the spheres at different positions along a tapered optical fiber, we show that the coupling constant exponentially increases with thinner fiber diameters. We demonstrate the close to critical coupling regime with intrinsic Q=3×10(4) for water immersed 14 µm BTG spheres.