LINC00205 modulates the expression of EPHX1 through the inhibition of miR-184 in hepatocellular carcinoma as a ceRNA.

Several studies have shown that low expression of epoxide hydrolase 1 (EPHX1) is closely associated with varying human cancers, including hepatocellular carcinoma (HCC). This study aims to explore the potential mechanism of EPHX1 silencing and revealed a novel regulatory pathway in the pathogenesis of HCC. In this study, micro ribonucleic acid (miR)-184 was predicted and validated to be a regulator of EPHX1 through experiments, and its expression was negatively correlated with the messenger RNA (mRNA) levels of EPHX1 in primary tumors. Elevation of EPHX1 suppressed cell proliferation and migration as well as cell cycle progression, and induced apoptosis, while downregulation of miR-184 exhibited the opposite effect on cellular processes. Moreover, LINC00205 interacted with miR-184 and was markedly downregulated in tumors. The effects of the miR-184 inhibitor on cell proliferation, apoptosis, and migration were reversed in part by the transfection with LINC00205 small interfering RNAs. In addition, LINC00205 acted as a molecular sponge to positively regulate the mRNA and protein levels of EPHX1 via regulating miR-184. The tumorigenicity of HCC cells was enhanced by LINC00205 shRNA but diminished by overexpression of EPHX1 in vivo. Clinically, the EPHX1 expression in patients with HCC was markedly downregulated. Taken together, the results of this study suggest that as a competing endogenous RNA, LINC00205 may regulate EPHX1 by inhibiting miR-184 in the progression of HCC and that targeting the LINC00205/miR-184/EPHX1 axis may provide a treatment protocol for patients.

MicroRNA-27a-3p inhibits cell viability and migration through down-regulating DUSP16 in hepatocellular carcinoma.

MicroRNA (miRNA or miR) has been shown to play an important role in the initiation and development in many different cancers. Here, we demonstrated down-regulated expression of miR-27a-3p in hepatocellular carcinoma (HCC) tissues in comparison with that in adjacent normal liver tissues based on the TCGA database. Cells viability and apoptosis was measured by CCK-8 and flow cytometry assay. Cell invasion and migration was measured by Transwell and wound healing assay. The effect of miR-27a-3p on DUSP16 expression was evaluated by luciferase assays, and western blot assay. miR-27a-3p up-regulation by transfection with miR-27a-3p mimics attenuated SMMC-7721 and HepG2 cell viability, invasion as well as migration, obviously. Moreover, we found that dual specificity phosphatase 16 (DUSP16), also known as mitogen-activated protein kinase phosphatase 7 (MKP-7), is a target of miR-27a-3p. DUSP16 expression was obvious decrease by miR-27a-3p at both transcriptional and protein levels in both SMMC-7721 and HepG2 cells. DUSP16 expression in tissues of HCC was up-regulated in comparison with that in tissues of adjacent liver based on the TCGA database. Overexpression of DUSP16 significantly reversed the cell changes in viability, invasion and migration which resulted from miR-27a-3p up-regulation in SMMC-7721 and HepG2 cells. Our findings contribute to current understanding of the functions of miR-27a-3p and suggest a mechanism by which miR-27a-3p plays an anti-tumor role in the development of HCC by targeting DUSP16.

G protein-coupled estrogen receptor enhances melanogenesis via cAMP-protein kinase (PKA) by upregulating microphthalmia-related transcription factor-tyrosinase in melanoma.

OBJECTIVE: This study investigated the role and mechanism of action of G protein-coupled estrogen receptor (GPER) in melanogenesis. METHODS: GPER expression was detected in the A375 human melanoma cell line and B16 mouse melanoma cell line. Cell proliferation, melanin content, tyrosinase (TYR) activity, cyclic adenosine monophosphate (cAMP) level, and TYR and microphthalmia-related transcription factor (MITF) expression were measured. GPER activation was altered by agonist and antagonist treatment and its expression was downregulated by gene silencing. Estradiol-induced melanin synthesis and the activation of related signaling pathways were suppressed by inhibiting GPER via antagonist treatment. The relationship between GPER and TYR was evaluated in clinical chloasma samples by immunohistochemistry. RESULTS: Upregulation of GPER in A375 cells promoted melanogenesis, favored as indicated by increases in TYR and MITF expression and TYR activity. GPER activated melanin production via the cAMP-protein kinase (PK) A pathway, suggesting that GPER plays an important role in estrogen-induced melanin synthesis. The effect of GPER activation on cAMP-MITF-TYR signaling was also demonstrated in B16 cells. A significant association was observed between GPER and TYR expression in chloasma skin lesions relative to normal skin. CONCLUSION: GPER enhances melanin synthesis via cAMP-PKA-MITF-TYR signaling and modulates the effects of estrogen in melanogenesis. GPER is therefore a potential drug target for chloasma treatment.

Spinal astrocytic activation contributes to mechanical allodynia in a rat chemotherapy-induced neuropathic pain model.

Chemotherapy-induced neuropathic pain (CNP) is the major dose-limiting factor in cancer chemotherapy. However, the neural mechanisms underlying CNP remain enigmatic. Accumulating evidence implicates the involvement of spinal glia in some neuropathic pain models. In this study, using a vincristine-evoked CNP rat model with obvious mechanical allodynia, we found that spinal astrocyte rather than microglia was dramatically activated. The mechanical allodynia was dose-dependently attenuated by intrathecal administratration of L-α-aminoadipate (astrocytic specific inhibitor); whereas minocycline (microglial specific inhibitor) had no such effect, indicating that spinal astrocytic activation contributes to allodynia in CNP rat. Furthermore, oxidative stress mediated the development of spinal astrocytic activation, and activated astrocytes dramatically increased interleukin-1ß expression which induced N-methyl-D-aspartic acid receptor (NMDAR) phosphorylation in spinal neurons to strengthen pain transmission. Taken together, our findings suggest that spinal activated astrocytes may be a crucial component of the pathophysiology of CNP and "Astrocyte-Cytokine-NMDAR-neuron" pathway may be one detailed neural mechanisms underlying CNP. Thus, inhibiting spinal astrocytic activation may represent a novel therapeutic strategy for treating CNP.

Anti-apoptotic effects of hyperoside via inhibition of NR2B-containing NMDA receptors.

Hyperoside (Hyp) is a flavonoid compound isolated from a folk remedy, Rhododendron ponticum L. leaves. It has been shown to have neuroprotective effects both in vivo and in vitro. However, little is known about the effects of Hyp on the neuronal apoptosis induced by glutamate. The present study showed that Hyp significantly attenuated, in a concentration-dependent manner, the apoptosis induced by the exposure of cultured neurons to NMDA. Western blot analysis revealed that Hyp antagonized the expression of excess NR2B-containing NMDA receptors; however, it had no effect on the expression of NR2A-containing NMDA receptors. Our results demonstrate that the neuroprotective effect of Hyp owes, at least partially, to its differential modulation of NR2A- and NR2B-containing NMDA receptors.

Hemodynamic features of Doppler ultrasonography in patients with portal hypertension: intraoperative direct measurement of portal pressure in the portal venous system.

OBJECTIVE: The purpose of this study was to investigate the relationship between a series of portal hemodynamic parameters obtained with Doppler ultrasonography and portal pressure measured directly from patients with portal hypertension (PHT). METHODS: Fifty-seven patients with a clinical diagnosis of PHT who accepted surgical therapy were investigated. The portal pressure was measured directly intraoperatively. Relevant parameters were compared and measured, including the hepatic artery pulsatility index (HAPI), hepatic artery resistive index (HARI), splenic artery resistive index, splenic artery pulsatility index (SpAPI), congestion index (CI) of the portal vein, hepatic buffer index (HBI), liver vascular index (LVI), and PHT index (PHI). RESULTS: Doppler parameters for the postprandial HAPI, SpAPI, CI, LVI, HBI, and PHI were statistically different in patients with PHT and healthy control subjects (P<0.05). The portal pressure was significantly correlated with the HARI (r=0.699; P<.001), HAPI (r=0.582; P<.001), LVI (r=-0.501; P=.003), HBI (r=0.441; P=.009), and Child-Pugh scores (r=0.589; P=.044). CONCLUSIONS: The HAPI, LVI, and HBI are indicative indices in patients with PHT, suggesting that color Doppler ultrasonography can be used as a noninvasive evaluation method for PHT degree. The changes in the HAPI, LVI, and HBI that accompany the increase in portal pressure can reflect hepatic resistance and hepatic artery buffer capacity accurately.

[Hyperthermia enhanced the killing effect of 5-fluorocytosine on human colon cancer cell line transfected with cytosine deaminase gene].

OBJECTIVE: To investigate whether hyperthermia can enhance the killing effect of 5- fluorocytosine (5- FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue- specific cytosine deaminase (CD) gene in vitro,and study its mechanism. METHODS: Human colorectal carcinoma cell lines SW480 transfected with G1CEACDNa were cultured. The proliferated colonies were treated with the combined therapy of 5-FC and hyperthermia at a temperature of 43 degrees C for 30 min. After eight days, MTT was used to calculate the cellular survival rate,to analyze the killing effect of 5-FC combined with hyperthermia on SW480 cells transfected with CD gene. Flow cytometry was performed to analyze the cellular cycle and transmission electron microscope was used to observe the morphologic changes of SW480 cells after thermochemotherapy. RESULTS: Hyperthermia combined with 5-FC had an enhanced killing effect on SW480-CEACD cells than 5-FC alone (P< 0.05, t =2.403, n=9). Flow cytometry revealed that the proportion of S stage cell increased in the group treated with hyperthermia and 5- FC (P< 0.001, t =7.158, n=6). Transmission electron microscope showed apoptosis after thermo- chemotherapy. CONCLUSIONS: Hyperthermia can improve the anti- tumor effect of 5- FC on human colorectal carcinoma cell lines SW480 transfected with CD gene, and the cells were blocked at S stage of cellular cycle and apoptosis was induced following thermochemotherapy.

[Effect of compound branch chain amino acids injection (Aminic) on protein metabolism in patients with radical resection of cardiac carcinoma].

OBJECTIVE: To evaluate the effect of compound branch chain amino acids injection (Aminic) on protein metabolism in patients with radical resection of cardiac carcinoma. METHODS: Eighty patients with radical resection of cardiac carcinoma were randomly divided into two groups:study group (40 cases) with compound branch chain amino acids injection (Aminic), and control group(40 cases) with amino acids(MPR- F). The patients received total parenteral nutrition with equal calorie and nitrogen from 1st-7th days after operation by central or peripheral vein with infusing time over 12 hours per day. The change of body weight was observed. The serum levels of total protein, albumin,pre-albumin,transferring, and fibronectin were measured. RESULTS: The body weight was decreased in the two groups after operation, while compared with the study group,the body weight was significantly decreased in the control group (P=0.0250). The positive nitrogen balance recovered two days earlier in study group than that in the control group. Decrease of total protein,albumin were more severe in the control group than those in study group (P=0.0446,P=0.0314 respectively). Compared with the control group,the level of Valine was increased (P=0.0073),and the level of Arginine was decreased more obviously in the study group (P= 0.0212). CONCLUSION: The compound branch chain amino acids injection (Aminic) is more beneficial for patients with radical resection of cardiac carcinoma to correct negative nitrogen balance,it can also inhibit decomposition of muscles protein,and improve nutritional conditions.

[Expression of matrix metalloproteinase 2 in oral verruvous carcinoma and squamous cell carcinoma].

OBJECTIVE: To determine the expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma. METHODS: Thirty cases were divided into 3 groups: verruvous carcinoma (n = 10), well-differentiated squamous cell carcinoma (n = 15) and moderately or poorly differentiated squamous cell carcinoma (n = 5). Reverse transcription polymerase chain reaction (RT-PCR) was used to test the expression of MMP2 mRNA in the carcinoma tissues and matched normal tissues from 3 groups above. RESULTS: The expression of MMP2 mRNA in the carcinoma tissues was significantly higher than that in their matched normal tissues (P < 0.05). The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P < 0.05). However, the expression of MMP2 mRNA was not obviously different between well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P > 0.05). CONCLUSION: The expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma tissues was significantly higher than that in their matched normal tissues. The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in squamous cell carcinoma.