Mitochondrial micropeptide MOXI promotes fibrotic gene transcription by translocation to the nucleus and bridging N-acetyltransferase 14 with transcription factor c-Jun.

Progressive fibrosis is a hallmark of chronic kidney disease, but we lack effective treatments to halt this destructive process. Micropeptides (peptides of no more than 100 amino acids) encoded by small open reading frames represent a new class of eukaryotic regulators. Here, we describe that the micropeptide regulator of ß-oxidation (MOXI) regulates kidney fibrosis. MOXI expression was found to be up-regulated in human fibrotic kidney disease, and this correlated with the degree of fibrosis and loss of kidney function. MOXI was expressed in the cytoplasm and mitochondria of cultured tubular epithelial cells and translocated to the nucleus upon Transforming Growth Factor-ß1 stimulation. Deletion of Moxi protected mice against fibrosis and inflammation in the folic acid and unilateral ureteral obstruction models. As a potential molecular therapy, treatment with an antisense MOXI oligonucleotide effectively knocked-down MOXI expression and protected against kidney fibrosis in both models. Bimolecular fluorescence complementation identified the enzyme N-acetyltransferase 14 (Nat14) and transcription factor c-Jun as MOXI binding partners. The MOXI/Nat14/c-Jun complex enhances basal and Transforming Growth Factor-ß1 induced collagen I gene promoter activity. Phosphorylation at T49 is required for MOXI nuclear localization and for complex formation with Nat14 and c-Jun. Furthermore, mice with a MoxiT49A point mutation were protected in the models of kidney fibrosis. Thus, our studies demonstrate a key role for the micropeptide MOXI in kidney fibrosis and identify a new function of MOXI in forming a transcriptional complex with Nat14 and c-Jun.

Imaging evaluation of para-aortic lymph nodes in cervical cancer.

BACKGROUND: In recent years, much literature has reported the diagnostic value of computed tomography (CT), magnetic resonance imaging (MRI), and positron emission tomography (PET)-CT in para-aortic lymph node metastasis of cervical cancer. PURPOSE: To compare and analyze the para-aortic lymph node presentations found in cervical cancer on different images in order to determine the best precise imaging method for identifying metastatic lymph nodes. MATERIAL AND METHODS: PubMed, Web of Science, MEDLINE, and other databases were searched for the non-invasive detection of metastatic lymph nodes for a comprehensive comparison. RESULTS: Positive lymph nodes on CT are significantly related to the following factors: short axis ≥10 mm; and round or central necrosis. Positive lymph nodes on MRI are significantly related to the following factors: short axis ≥8 mm; inhomogeneous signal intensity; morphology: round, irregular edge, extracapsular invasion, central necrosis, loss of lymph node structure, burrs, or lobes; and ADC value decreases, combined with local actuality. On PET-CT examination, when the short axis of the lymph node is >5 mm, the SUV is >2.5, or the FDG uptake is greater than that of the surrounding tissue, it is a metastatic lymph node. CONCLUSION: In conclusion, different imaging techniques show metastatic lymph nodes in different ways. Combining the patient's medical history with the symptoms of the aforementioned lymph nodes, together with one or more imaging techniques, is important to diagnose para-aortic lymph nodes in cervical cancer.

Efficacy and tolerability of docetaxel and cisplatin plus S-1 for advanced gastric cancer.

PURPOSE: The aim of this study was to evaluate the efficacy and tolerability of docetaxel and cisplatin plus S-1 (DCS) combination chemotherapy in advanced gastric cancer patients. METHODS: Chemo-naive patients with advanced gastric cancer, ECOG performance status of 0 to 1, and adequate organ function were eligible. All patients received docetaxel 75 mg/m(2) and cisplatin 75 mg/m(2) on day 1, plus S-1 orally 40-60 mg bid depending on body surface area on days 1-14, every 21 days. Efficacy and adverse events were evaluated every two cycles. RESULTS: Fifty-nine patients were enrolled from February 2009 to January 2011 and 56 of them were evaluated for efficacy and tolerability. After a median follow up of 17.6 months, the objective response rate (RR) was 75%, the disease control rate (DCR) 83.9%, the median progression free survival (PFS) and overall survival (OS) 6.5 (95% CI, 5.6-7.3) months and 15.5 (95% CI, 13.9-17.0) months, respectively. The median number of chemotherapy cycles was 5. Grade 3 or 4 adverse effects included neutropenia (60.7%), vomiting (14.3%), neurotoxicity (12.5%), thrombocytopenia (10.7%), diarrhea (10.7%), impaired liver function (3.6%), and hand-foot syndrome (1.8%). CONCLUSION: Our study shows that DCS regimen is active against advanced gastric cancer with acceptable toxicities and it may be used as a new choice of first-line chemotherapy for patients with advanced gastric cancer.

Bisphosphonates and risk of lung cancer: Protocol for a systematic review and meta-analysis.

ABSTRACT: The association between the use of bisphosphonates (BPs) and the risk of lung cancer has been concerned recently. There is no explicit study indicating that whether the use of BPs would affect the risk of lung cancer. So, we conducted a meta-analysis to figure out the relationship between BPs and lung cancer.We searched the databases of PubMed and Embase. The random effects were used to calculate the pooled odds ratios (ORs) and 95% confidence interval (CIs) for the risk of lung cancer in BPs users compared with non-users. The stability of our results was evaluated by the sensitivity analysis. The publication bias was assessed in our study. The data in our study comes from the public database, therefore ethical approval is not necessary. Also, our study did not involve patient consent.Four studies met our inclusion criteria. All the included studies are cohort studies. Our analysis indicated that there was no significant association between the use of BPs and the risk of lung cancer (OR 1.02, 95%CI 0.85- 1.24, I2 71%). In our secondary analysis, the use of alendronate may increase the risk of lung cancer. The pooled OR of 3 studies is (OR 1.10, 95%CI 0.84-1.45, I2 77%), but when we performed a sensitivity analysis, 1 of the OR is (OR 1.23, 95%CI 1.02-1.49, I2 4.1%).This is the most detailed meta-analysis on this topic. And there was no significant association between the use of BPs and lung cancer. However, exposure to alendronate may increase the risk of lung cancer. More studies are needed to confirm our findings.

miR-30b-5p releases HMGB1 via UBE2D2/KAT2B/HMGB1 pathway to promote pro-inflammatory polarization and recruitment of macrophages.

BACKGROUND AND AIMS: Oxidized low-density lipoprotein (ox-LDL) is a key risk factor for atherosclerosis, but there are few reports on ox-LDL-mediated inflammation after injury. In this study, we investigated the effects of ox-LDL on endothelial injury and macrophage polarization and recruitment using human aortic endothelial cell line HAEC cells. METHODS: Changes in miRNA levels after ox-LDL treatment were assessed with qRT-PCR. Luciferase experiments were performed to verify the interaction between miRNA and protein, and co-IP and ubiquitination experiments to detect proteins interactions. Cell phenotype was assessed by cytometry and Western blot. RESULTS: qRT-PCR data indicated that ox-LDL treatment up-regulates the expression of miR-30b-5p. Luciferase test and ubiquitination assay showed miR-30b-5p can bind to UBE2D2 and reduce its ubiquitination ability to degrade KAT2B. The up-regulated KAT2B promotes the acetylation of HMGB1, acetylated HMGB1 dissociates from SIRT1, exit the nucleus, and it is secreted from the cell. Flow cytometry and transwell experiments showed that HMGB1 secreted from HAEC can induce pro-inflammatory (M1-like) polarization and recruitment of RAW264.7 cells. CONCLUSIONS: Our results indicate that ox-LDL activates the UBE2D2/KAT2B pathway by upregulating miR-30b-5p, thus acetylating HMGB1, which is then secreted from the cell, thereby promoting pro-inflammatory cell polarization and recruitment of macrophage.

MicroRNA-494-3p Promotes Cell Growth, Migration, and Invasion of Nasopharyngeal Carcinoma by Targeting Sox7.

BACKGROUND: There is mounting evidence that microRNAs play an important role in nasopharyngeal carcinoma, which is widely prevalent in South China and is the most prevalent metastatic cancer among head and neck cancers. Recently, it has been shown that miR-494 is involved in the progression and prognosis of nasopharyngeal carcinoma. However, little is known about the function and mechanism of miR-494-3p in nasopharyngeal carcinoma. In the present study, we aimed to investigate the effects of miR-494-3p on the migration and invasion of nasopharyngeal carcinoma and to further explore the underlying mechanisms of these processes. METHODS: The expression levels of miR-494-3p and Sox7 in nasopharyngeal carcinoma specimens and nasopharyngeal carcinoma cell lines were measured using quantitative reverse transcription polymerase chain reaction. Luciferase reporter assay, quantitative reverse transcription polymerase chain reaction, and Western blotting were used to confirm whether Sox7 was a direct target of miR-494-3p. Additionally, the roles of miR-494-3p and Sox7 on cell proliferation, migration, and invasion of nasopharyngeal carcinoma were analyzed by Cell Counting Kit-8 (CCK-8) assay, wound healing assay, and Boyden chamber assay, respectively. RESULTS: Our study demonstrated that miR-494-3p was commonly upregulated in nasopharyngeal carcinoma specimens and nasopharyngeal carcinoma cell lines compared with nontumor nasopharyngeal epithelial tissue or nasopharyngeal cells (NP69). Moreover, miR-494-3p negatively regulated Sox7 at the posttranscriptional level by binding to a specific site in the Sox7 3'-untranslated region. In addition, synthetic miR-494-3p mimics significantly promoted proliferation, migration, and invasion of S18 and S26 nasopharyngeal carcinoma cells, while a synthetic miR-494-3p inhibitor resulted in suppressed nasopharyngeal carcinoma cell migration and invasion. CONCLUSION: miR-494-3p promotes nasopharyngeal carcinoma cell growth, migration, and invasion by directly targeting Sox7. Our results suggest that miR-494-3p might be a potential therapeutic target for nasopharyngeal carcinoma.

Matrine suppresses the migration and invasion of NSCLC cells by inhibiting PAX2-induced epithelial-mesenchymal transition.

Non-small cell lung cancer (NSCLC) is the major cause of deaths among all the cancer types worldwide. Most of the NSCLC is diagnosed at an advanced stage and the 5-year overall survival rate is low. The reason for the low survival rate of patients with NSCLC is mainly due to distant metastasis. Matrine, a traditional Chinese medicine, has been shown a significant anti-proliferation and anti-invasive effect in tumors. However, little is known on the anti-invasive mechanism of matrine in lung cancer. Therefore, we tried to investigate the molecular mechanism of matrine on the invasive ability of NSCLC cells in vitro. Cell Counting Kit-8 assay was used to evaluate the cell viability. Transwell assay was used to detect the migration and invasion abilities. Microarray assay was used to analyze the differentiated expression genes with or without matrine treatment. Western blotting and real-time polymerase chain reaction were applied to detect the expressions of PAX2, E-cadherin and N-cadherin. Our study showed that matrine could suppress the proliferative activity of NSCLC cells in a dose- and time-dependent manner. Further investigation discovered that the migration and invasion of NSCLC cells were significantly inhibited by treatment with different concentrations of matrine. Microarray assay, real-time polymerase chain reaction and western blotting showed that matrine could significantly decrease the expression of PAX2. In addition, epithelial-mesenchymal transition and related proteins were decreased. In conclusion, matrine may block PAX2 expression to interfere with epithelial-mesenchymal transition signaling pathway that ultimately inhibit the migration and invasion of NSCLC cells in vitro. Matrine might serve as a potential agent for NSCLC treatment.

[Clinicel study on treatment of advanced primary liver cancer by Yanshu injection combining with chemotherapy].

OBJECTIVE: To study the effects of Yanshu injection on the combined treatment in the advanced primary liver cancer. METHOD: Eighty-five cases of advanced primary liver cancer were treated with Yanshu injection combining with chemotherapy or only chemotherapy. The curative effects, pain genesic rate, one year survival rate, survival quality of life and cell immune functions were observed. RESULT: The remission rate and one year survival rate of the trial group were 60.5% and 51.2%, respectively, and were significantly higher than those (45.2% and 40.5%) of the control group (P < 0.05). The pain relief rate of the trial group was significantly higher than that of the control group (P < 0.05). The improvement of the quality of life was higher than that of the contral group (P < 0.01). The ability of the T-cell subgroup and NK-cell of the trail group were significantly difference between pre-and post-treatment (P < 0.01 or 0.05); however, that of the control group was no obviously change. CONCLUSION: Yanshu injection combination with chemotherapy can raise the curative effect, one year survival rate and cellular immune function, reduce pain genesic rate and toxicity of chemotherapy, and improve the quality of life of the patients with advanced primary liver cancer, which is worthy to be recommended for clinical application.

Clinical significance of microRNA-155 expression in hepatocellular carcinoma.

The present study aimed to evaluate the expression of microRNA-155 (miR-155) in hepatocellular carcinoma (HCC) and adjacent normal tissues, and assess its correlation with clinicopathological characteristics of this tumor type. miR-155 expression was detected in 40 HCC tissue samples and 40 samples of adjacent tumor-free tissue using fluorescent reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The association between miR-155 expression, clinicopathological features and 1-year relapse-free survival (RFS) in HCC and adjacent normal tissue samples was analyzed. RT-qPCR results revealed that, in 25 cases (62.5%), miR-155 expression levels were significantly increased in HCC tissues compared with the expression levels observed in pericarcinomatous tissues (P<0.05). miR-155 expression was observed to be significantly correlated with vessel invasion, Edmonson classification and clinical stage (P<0.05). However, miR-155 expression was not significantly correlated with gender, age, tumor size, tumor number, hepatitis B virus DNA copy number, cirrhosis or concentration of α-fetoprotein (P>0.05). A positive correlation was observed between late TNM classification of malignant tumor stage and 1-year RFS (P<0.05). Patients exhibiting high miR-155 expression levels were observed to exhibit a lower 1-year RFS than that of patients with reduced expression of miR-155 (48 vs. 73.3%), however this difference was not statistically significant (P=0.105). Additionally, correlations were observed between miR-155 expression and reduced differentiation, increased invasiveness and late stages of HCC. The current results demonstrated that miR-155 may be involved in the tumorigenesis of HCC and may be associated with clinical characteristics of HCC patients. Additional studies are required to clarify the mechanism of miR-155.

[Synergistic Antitumor Effect of Amorphigenin Combined with Cisplatin in Human Lung Adenocarcinoma A549/DDP Cells].

BACKGROUND: Amorphigenin, a rotenoid compouns, from seeds of Amorpha fruticosa, has been shown to possess anti-proliferation activities in several cancer cells. To explore the antitumor effects of amorphigenin on cisplatin-resistant human lung adenocarcinoma A549/DDP cells and explore the underlying mechanisms. METHODS: CCK-8 assay was used to measure the proliferation of A549/DDP cells; Colony formation assay was used to measure the colony formation of A549/DDP cells; Flow cytometry assay was used to detect the apoptosis rates; Western blot analysis was used to explore the expression of apoptosis-related proteins (caspase-3 protein, PARP protein) and lung resistance protein (LRP). RESULTS: Our results demonstrated that amorphigenin could inhibit the proliferation of A549/DDP cells with a inhibition concentration of 50% cell growth (IC50) at 48 h of (2.19±0.92) µmol/L. Amorphigenin could inhibit the colony formation ability and induce apoptosis of A549/DDP cells; Furthermore, amorphigenin combined with cisplatin showed synergistic proliferation-inhibitory effect and apoptosis-promoting effect in A549/DDP cells; reduced the expression of LRP of A549/DDP cells. CONCLUSIONS: Amorphigenin remarkably inhibits the proliferation and induces apoptosis in A549/DDP cells. Combination of amorphigenin with cisplatin had the synergistic inhibitory effect on A549/DDP cells by downregulating the expression of LRP.
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Clinical significance of axin and ß-catenin protein expression in primary hepatocellular carcinomas.

The aim of the present research was to investigate clinicopathologic correlations of immunohistochemically- demonstrated axin (axis inhibition) and ß-catenin expression in primary hepatocellular carcinomas (HCCs), in comparison with paraneoplastic, cirrhotic and normal liver tissues. Variation in Axin expression across groups were significant (P < 0.01), correlating with alpha fetoprotein (AFP), HBsAg, cancer plugs in the portal vein, and clinical stage of HCCs(P < 0.05); however, there were no links with sex, age, and tumour size (P > 0.05). Differences in cell membrane ß-catenin expression were also statistically significant (P < 0.01), again correlated with AFP, HBsAg, cancer plugs in the portal vein, and clinical stage in HCCs (P < 0.05) but not with sex, age, and tumour size (P > 0.05). Axin expression levels in tissues with reduced membrane ß-catenin were low (P < 0.05), also being low with nuclear ß-catenin expression (P < 0.05). Axin and ß-catenin may play an important role in the genesis and progression of HCC via the Wnt signal transmission pathway. Simultaneous determination of axin, ß-catenin, AFP, and HBsAg may be useful for early diagnosis, and metastatic and clinical staging of HCCs.

DLC-1 expression levels in breast cancer assessed by qRT- PCR are negatively associated with malignancy.

OBJECTIVE: The aim of this study was to explore the expression of DLC-l in breast carcinoma and any association with tumor metastasis. METHODS: 51 surgical specimens of human breast carcinoma, divided into high invasive and low invasive groups according to their clinicopathological features, 30 cases of adjacent normal tissue and 28 benign breast lesions were examined by qRT-PCR for expression of DLC-1. RESULTS: Expression level of DLC-1 in adjacent normal tissue and benign breast lesion specimens was higher than that in breast carcinoma (P<0.0001); the values in the high invasive group with synchronous metastases were also lower than in the low invasive group (P=0.0275). The correlation between DLC-1 expression level and tumor progression and metastasis of breast cancer was negative. CONCLUSION: As an anti-oncogene, DLC-1 could play an important part in breast carcinoma occurrence, progression, invasiveness and metastasis. Detecting the changes of the expression of DLC-1 in the breast carcinoma may contribute to earlier auxiliary diagnosis of invasiveness, metastasis and recrudescence.