Raloxifene Protects Cisplatin-Induced Renal Injury in Mice via Inhibiting Oxidative Stress.

PURPOSE: Cisplatin is one of the most widely used antineoplastic drugs but has limited therapeutic effects due to nephrotoxicity. The aim of this study was to determine the possible renoprotective effect of the antioxidant raloxifene on cisplatin-induced nephrotoxicity in mice. MATERIALS AND METHODS: Cisplatin-induced acute renal injury was established in female C57 mice that were treated with saline (normal control) or raloxifene over a 7-day period. The body weight of the mice was recorded. Histopathological examinations of the kidney tissues were performed using H&E, PAS staining and TEM. The histomorphology of liver and other organs was observed by H&E staining. The serum levels of creatinine, blood urea nitrogen (BUN), alanine transaminase (ALT) and glutamic oxalacetic transaminase (AST) were analyzed by specific kits. Superoxide dismutase (SOD) and glutathione (GSH) activity, and the content of malondialdehyde (MDA) in the kidney, liver homogenates and HK-2 cells were measured by WST-8 and thiobarbituric acid colorimetric methods. Moreover, the mitochondrial structures of HK-2 cells were performed using TEM. The viability and proliferation of HK-2 cells were examined by CCK-8 and EdU incorporation assays. The mitochondrial membrane potential was measured by JC-1 fluorescence. RESULTS: Raloxifene significantly reduced the levels of serum creatinine, urea, ALT and AST in the cisplatin-treated mice, and alleviated cisplatin-induced renal and hepatic tissue injury. Furthermore, raloxifene also increased the activity of GSH and SOD in the renal tissues and HK-2 cells, and reduced MDA levels, thereby limiting oxidative stress in the kidney. CONCLUSION: Raloxifene protected against cisplatin-induced nephrotoxicity by activating the antioxidant system, along with alleviating liver damage. It should be considered as a potential adjuvant in cisplatin-based chemotherapeutic protocols.

Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-ß1/STAT3 Axis in vitro.

AIM: Basal epithelial cells are absent in distant prostate cancer. This study aimed to investigate whether basal epithelial cells could suppress migration and invasion of prostate cancer cells to become a new treatment strategy for prostate cancer. MAIN METHODS: Basal epithelial cells were identified by immunofluorescence with anti-p63. Wound healing assays or transwell assays were used to explore the effects of basal epithelial cells, TGF-ß1, SB431542 (inhibitor of TGF-ß type I receptor) or stattic (inhibitor of phosphorylated STAT3) on migration or invasion of mouse prostate cancer cell (RM-1). Concentration of TGF-ß1 was measured by ELISA assay. HE staining was used to investigate cell morphology. Immunocytochemistry with anti-p63 was used to identify basal epithelial cells. Levels of STAT3, p-STAT3 (Ser727) and proteins associated with EMT were measured with Western blot assay. Cell proliferation was measured with MTT or CCK8 assay. RESULTS: Normal basal epithelial cells acquired from mouse prostate were specific to anti-p63 and more than 90%. Basal epithelial cells and RM-1 could both secrete TGF-ß1. Basal epithelial cells and TGF-ß1 promoted the migration and invasion of RM-1 through changing the cell morphology and up-regulating expression of ZEB1, N-cadherin, vimentin, snail and p-STAT3 (Ser727), at the same time down-regulating E-cadherin of RM-1. SB431542 strongly suppressed migration, invasion as well as the expressions of EMT relevant proteins and p-STAT3 (Ser727) of co-cultured RM-1. In addition, stattic suppressed proliferation, migration and invasion of non-treated RM-1 and co-cultured RM-1. CONCLUSION: Our study suggests that normal basal epithelial cells might stimulate the migration and invasion of RM-1 by TGF-ß1/STAT3 axis which could be suppressed by inhibitor of TGF-ß receptor and inhibitor of p-STAT3. So, basal epithelial cells might not become a treatment strategy for prostate cancer, but our results could provide some researching references for other diseases which include basal epithelial cells such as prostatic intraepithelial neoplasia, prostatic hyperplasia, cervical cancer, or urinary bladder cancer.

Endometrial clear cell carcinoma invading the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor: A case report.

BACKGROUND: To investigate the clinicopathological features of endometrial clear cell carcinoma that has invaded the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor. CASE SUMMARY: A case of endometrial clear cell carcinoma invading the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor was collected and analyzed using pathomorphology and immunohistochemistry. Endometrial clear cell carcinoma cells were distributed in a solid nest, papillary, shoe nail-like, and glandular tube-like distribution. There was infiltrative growth, and tumor cells had clear cytoplasm and obvious nuclear heteromorphism. The cancer tissue was necrotic and mitotic. The cancer tissue invaded the right oviduct. The ipsilateral oviduct also had an adenomatoid tumor. The adenomatoid tumor was arranged in microcapsules lined with flat or cubic cells that were surrounded by smooth muscle tissue. The adenomatoid tumor cells were round in shape. CONCLUSION: Clear cell carcinoma of the endometrium can invade the oviduct and occur simultaneously with tubal adenomatoid tumors. Upon pathological diagnosis, one should pay close attention to distinguishing whether an endometrial clear cell carcinoma is invading the oviduct or whether it is accompanied by an adenomatoid tumor of the oviduct. Immunohistochemistry is helpful to differentiate these two disease entities. Endometrial clear cell carcinomas express Napsin-A and P16 and are negative for estrogen receptor and progesterone receptor. The presence of endometrial clear cell carcinoma does not affect the expression of CK and calretinin in adenomatoid tumors.

The clinicopathologic and immunohistochemical features of villoglandular adenocarcinoma of uterine cervix.

AIM: Villoglandular adenocarcinoma (VGA) of the uterine cervix is a variant of endocervical adenocarcinoma. However, the clinicopathologic and immunohistochemical features of VGA are still unclear. The aim of this study was to investigate the clinicopathologic and immunohistochemical features of VGA. MATERIALS AND METHODS: A total of 20 VGA patients were identified among 852 patients diagnosed with cervical cancer and enrolled in this study. The immunohistochemical levels of Ki-67, P53, P16, progesterone receptor (PR), carcinoembryonic antigen (CEA), vimentin (Vim), and estrogen receptor (ER) were measured by immunohistochemistry. RESULTS: VGA was prevalent in younger women and presented favorable prognosis. Ki-67, P16, and CEA were highly expressed in VGA tissues, while PR expression was hardly to be detected. The positive rates of Ki-67, CEA, and P16 were 90.0%, 90.0%, and 85.0%, respectively, which were significantly higher compared with PR (5.0%, P < 0.001). In addition, the positive rates of P53, Vim, and ER in VGA tissues were 55.0%, 50.0%, and 40.0%, respectively. However, the expression levels of Ki-67, P53, P16, PR, CEA, Vim, and ER were not significantly associated with clinical features (P > 0.05). CONCLUSION: These data indicate that VGA is a rare cervical adenocarcinoma, which is prevalent in younger women, and presents favorable prognosis. Detection of Ki-67, P53, P16, PR, CEA, Vim, and ER would be beneficial for the diagnosis of VGA.

[Prognostic value of circulating tumor cells and disseminated tumor cells in patients with esophageal cancer: a meta-analysis].

OBJECTIVE: To explore the correlations of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) with the clinicopathological characteristics, prognostic events, and survival outcomes in esophageal cancer (EC) patients. METHODS: The PubMed, Web of Science, Embase database and Cochrane database were searched for studies reporting the outcomes of interest. The studies were selected according to established inclusion/exclusion criteria. Meta-analysis of the studies was performed using Review Manager 5.3 and Stata12.0 software with the odds ratio (OR), risk ratio (RR) , hazard ratio (HR) , and 95% confidence interval (95% CI) as the effect indexes. RESULTS: Nineteen studies involving a total of 1766 patients were included in the analysis. Significant correlations of CTCs and DTCs were found with the clinicopathological parameters including the tumor stage (OR=1.95), depth of invasion (OR=1.99), lymph node metastasis (OR=2.44SEN), distal metastasis (OR=5.98SEN), histological differentiation (OR=1.67) and lymphovascular invasion (OR=4.48). CTCs and DTCs were also correlated with the prognostic events including relapse (RR=6.86SEN) and metastasis (RR=3.22) and with the survival outcomes including the overall survival (OS) overall analysis (HR=3.46) and disease-free survival/progression-free survival (DFS/PFS) overall analysis (HR=3.00). CONCLUSION: CTCs and DTCs are significantly associated with an advanced tumor stage, depth of tumor invasion, lymph node metastasis, distant metastasis before therapy, differentiation, lymphovascular invasion, relapse and metastasis in patients with EC. They are also significantly correlated with a poorer survival for OS and DFS/PFS to serve as clinical and prognostic predictors in patients with EC.

Plasma glutamate-modulated interaction of A2AR and mGluR5 on BMDCs aggravates traumatic brain injury-induced acute lung injury.

The bone marrow-derived cell (BMDC)-associated inflammatory response plays a key role in the development of acute lung injury (ALI). Activation of adenosine A2A receptor (A2AR) is generally considered to be antiinflammatory, inhibiting BMDC activities to protect against ALI. However, in the present study, we found that in a mouse model of neurogenic ALI induced by severe traumatic brain injury (TBI), BMDC A2AR exerted a proinflammatory effect, aggravating lung damage. This is in contrast to the antiinflammatory effect observed in the mouse oleic acid-induced ALI model (a nonneurogenic ALI model.) Moreover, the A2AR agonist CGS21680 aggravated, whereas the antagonist ZM241385 attenuated, the severe TBI-induced lung inflammatory damage in mice. Further investigation of white blood cells isolated from patients or mouse TBI models and of cultured human or mouse neutrophils demonstrated that elevated plasma glutamate after severe TBI induced interaction between A2AR and the metabotropic glutamate receptor 5 (mGluR5) to increase phospholipase C-protein kinase C signaling, which mediated the proinflammatory effect of A2AR. These results are in striking contrast to the well-known antiinflammatory and protective role of A2AR in nonneurogenic ALI and indicate different therapeutic strategies should be used for nonneurogenic and neurogenic ALI treatment when targeting A2AR.

Adenosine A2A receptors in both bone marrow cells and non-bone marrow cells contribute to traumatic brain injury.

Adenosine A2A receptors (A(2A)Rs) in bone marrow-derived cells (BMDCs) are involved in regulation of inflammation and outcome in several CNS injuries; however their relative contribution to traumatic brain injury (TBI) is unknown. In this study, we created a mouse cortical impact model, and BMDC A(2A)Rs were selectively inactivated in wild-type (WT) mice or reconstituted in global A(2A)R knockout (KO) mice (i.e. inactivation of non-BMDC A(2A)Rs) by bone marrow transplantation. When compared with WT mice, selective inactivation of BMDC A(2A)Rs significantly attenuated the neurological deficits, brain water content and cell apoptosis at 24 h post-TBI as global A(2A)R KO did. However, compared with the A(2A)R KO mice, selective reconstitution of BMDC A(2A)Rs failed to reinstate brain injury, indicating the contribution of the non-BMDC A(2A)R to TBI. Furthermore, the protective outcome by selective inactivation of BMDC A(2A)R or broad inactivation of non-BMDC A(2A)Rs was accompanied with reduced CSF glutamate level and suppression of the inflammatory cytokines interleukin-1, or interleukin-1 and tumor necrosis factor-alpha. These findings demonstrate that inactivation of A(2A)Rs in either BMDCs or non-BMDCs is sufficient to confer the protective effect as global A(2A)R KO against TBI, indicating the A(2A)R involvement in TBI by multiple cellular mechanisms of A(2A)R involvement including inhibition of glutamate release and inflammatory cytokine expressions.

Local glutamate level dictates adenosine A2A receptor regulation of neuroinflammation and traumatic brain injury.

During brain injury, extracellular adenosine and glutamate levels increase rapidly and dramatically. We hypothesized that local glutamate levels in the brain dictates the adenosine-adenosine A(2A) receptor (A(2A)R) effects on neuroinflammation and brain damage outcome. Here, we showed that, in the presence of low concentrations of glutamate, the A(2A)R agonist 3-[4-[2-[[6-amino-9-[(2R,3R,4S,5S)-5-(ethylcarbamoyl)-3,4-dihydroxy-oxolan-2-yl]purin-2-yl]amino]ethyl]phenyl]propanoic acid (CGS21680) inhibited lipopolysaccharide (LPS)-induced nitric oxide synthase (NOS) activity of cultured microglial cells, an effect that was dependent on the protein kinase A (PKA) pathway. However, in high concentrations of glutamate, CGS21680 increased LPS-induced NOS activity in a protein kinase C (PKC)-dependent manner. Thus, increasing the local level of glutamate redirects A(2A)R signaling from the PKA to the PKC pathway, resulting in a switch in A(2A)R effects from antiinflammatory to proinflammatory. In a cortical impact model of traumatic brain injury (TBI) in mice, brain water contents, behavioral deficits, and expression of tumor necrosis factor-alpha, interleukin-1 mRNAs, and inducible NOS were attenuated by administering CGS21680 at post-TBI time when brain glutamate levels were low, or by administering the A(2A)R antagonist ZM241385 [4-(2-{[5-amino-2-(2-furyl)[1,2,4]triazolo[1,5-a][1,3,5]triazin-7-yl]amino}ethyl)phenol] at post-TBI time when brain glutamate levels were elevated. Furthermore, pre-TBI treatment with the glutamate release inhibitor (S)-4C3HPG [(S)-4-carboxy-3-hydroxyphenylglycine] converted the debilitating effect of CGS21680 administered at post-TBI time with high glutamate level to a neuroprotective effect. This further indicates that the switch in the effect of A(2A)R activation in intact animals from antiinflammatory to proinflammatory is dependent on glutamate concentration. These findings identify a novel role for glutamate in modulation of neuroinflammation and brain injury via the adenosine-A(2A)R system.

[Establishment of a selective inactivation adenosine A2A receptors mice model].

OBJECTIVE: To establish a mice model with selective inactivation adenosine A2A receptors (A2ARs) in peripheral white blood cells (PWBC). METHODS: A2ARs were selectively inactivated in PWBCs by transplanting bone marrow cells (BMCs) from A2AR knockout (KO) mice into their wild type (WT) littermates after a single total body irradiation of 9.5 Gy or fractionated total body irradiation of 6.2 Gy x 2. The efficiency of reconstitution of bone marrow-derived cells in chimeric mice was assessed. RESULTS: PCR band patterns changed from the recipient pattern (one band of 330 bp) to the donor (two bands of 300 and 330 bp) pattern. Immunohistochemistry analysis showed that 10.21% of cells were A2AR+ in PWBCs in KO--> WT mice, whereas 96.72% of cells were A2AR+ in WT mice. The survival rates of mice irradiated with 6.2 Gy x 2 and transplanted with more than 6 x 10(6) BMCs were about 91%. CONCLUSION: A murine model of selective inactivation adenosine A2A receptors in PWBCs was established successfully.

[Expression of dopamine receptor D2 and adenosine receptor A2A in human retinal pigment epithelium].

OBJECTIVE: The aim of this study was to identify the presence of dopamine receptor D2 and adenosine receptor A2A in human retinal pigment epithelium (RPE) in order to determine whether the RPE is a potential site of action for these two receptors. METHODS: RPE Cells were isolated and cultured. Reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry were performed to detect the expression of D2 and A2A receptors in the RPE. RESULTS: RT-PCR analysis showed that mRNA of D2 and A2A receptors were expressed in the RPE. D2 and A2A receptors were distributed in the RPE by immunocytochemistry. CONCLUSIONS: This study demonstrated the presence of D2 and A2A receptors in human RPE at both mRNA and protein levels. D2 and A2A receptors may play an important role in the maintenance of the function of human retinal pigment epithelium. Furthermore, their cooperation appeared to be important in the development of axial myopia and retinal and choroidal neovascularization.