HBV preS2 promotes the expression of TAZ via miRNA-338-3p to enhance the tumorigenesis of hepatocellular carcinoma

Transactivators encoded by HBV, including HBx and preS2, play critical role in hepatocellular carcinoma (HCC). YAP, a downstream effector of the Hippo pathway, is involved in hepatocarcinogenesis mediated by HBx. Here, we investigated whether preS2, another transactivator encoded by HBV, regulates the Hippo pathway to promote HCC. We found that preS2 overexpression upregulated TAZ, a downstream effector of the Hippo pathway, at protein level but not at mRNA level. preS2 suppressed miRNA-338-3p expression in HCC cell lines. miRNA-338-3p mimics downregulated TAZ, while miRNA-338-3p inhibitor restored the expression of TAZ, suggesting that TAZ is a direct target of miRNA-338-3p. TAZ overexpression stimulated growth of HCC cell lines. Knockdown of TAZ dampened preS2-promoted HCC proliferation and migration. Thus, preS2 upregulates TAZ expression by repressing miRNA-338-3p. TAZ is necessary for preS2-promoted HCC proliferation and migration     

miR-9-3p plays a tumour-suppressor role by targeting TAZ (WWTR1) in hepatocellular carcinoma cells

Background:

The inactivation of the Hippo pathway lead to TAZ (PDZ-binding motif)/YAP (yes-associated protein) overexpression, and is associated with worse prognostic outcomes in various cancers including hepatocellular carcinoma (HCC). Although there are several reports of microRNA (miR) targeting for YAP, miR targeting for TAZ remains unclear. The aim of this study is to identify the miR targeting TAZ expression in HCC.

Methods:

MicroRNA expression was analysed using the Human miFinder 384HC miScript miR PCR array, and was compared between low and high TAZ expression cell lines. Then, we extracted miR-9-3p as a tumour-suppressor miR targeting TAZ. We examined the functional role of miR-9-3p using miR-9-3p mimic and inhibitor in HCC cell lines).

Results:

In HCC cell lines and HCC clinical samples, there was the inverse correlation between miR-9-3p and TAZ expressions. TAZ expression was induced by treatment of miR-9-3p inhibitor and was downregulated by treatment of miR-9-3p mimic. Treatment of miR-9-3p mimic inhibited cell proliferative ability with downregulated phosphorylations of Erk1/2, AKT, and β-catenin in HLF. Inversely, treatment of miR-9-3p inhibitor accelerated cell growth compared with control in HuH1.

Conclusions:

MicroRNA-9-3p was identified as the tumour-suppressor miR targetting TAZ expression in HCC cells.     

Antimicrobial prophylaxis for transurethral resection of bladder tumor: A retrospective comparison of preoperative single-dose administration of piperacillin and tazobactam/piperacillin

We aimed to clarify prophylactic antimicrobial effects of single-dose piperacillin (PIPC) for perioperative infections in the transurethral resection of bladder tumor (TURBT) in comparison with those of single-dose tazobactam/piperacillin (TAZ/PIPC) through a retrospective analysis. We analyzed data from 192 TURBT patients treated with single-dose (4 g) intravenous PIPC (P group) between April 2015 and April 2017. For comparison, we analyzed data from 50 TURBT patients treated with single-dose (4.5 g) intravenous TAZ/PIPC (T/P group) between June 2013 and April 2014. We compared the perioperative incidences of fever (≥38 °C) and bacteriuria in the two groups. The number of febrile patients was four (2.1%) in the P group and one (2.0%) in the T/P group, without significant difference (p = 0.970). Among these febrile patients, urine and blood samples of two patients in the P group tested positive for bacterial cultures of Citrobacter koseri and Enterococcus faecalis, respectively. None of the patients in the T/P group tested positive for urine culture, postoperatively. However, 22 patients (18.2%) in the P group tested positive for urine culture, and Staphylococcus epidermidis (six patients), E. faecalis (three patients), Escherichia coli (three patients), Streptococcus agalactiae (two patients), Staphylococcus aureus (two patients), and C. koseri (one patient) were isolated. There was no significant difference in the incidence of bacteriuria in these two groups (p = 0.055). Based on these results, single-dose PIPC administration for the prevention of perioperative infections in TURBT was as effective as TAZ/PIPC.     

Transport characteristics and transporter‐based drug–drug interactions of TM‐25659, a novel TAZ modulator

The in vitro metabolic stability and transport mechanism of TM‐25659, a novel TAZ modulator, was investigated in human hepatocytes and human liver microsomes (HLMs) based on the preferred hepatobiliary elimination in rats. In addition, the in vitro transport mechanism and transporter‐mediated drug–drug interactions were evaluated using oocytes and MDCKII cells overexpressing clinically important drug transporters. After a 1 h incubation in HLMs, 92.9 ± 9.5% and 95.5 ± 11.6% of the initial TM‐25659 remained in the presence of NADPH and UDPGA, respectively. Uptake of TM‐25659 readily accumulated in human hepatocytes at 37 ºC (i.e. 6.7‐fold greater than that at 4 ºC), in which drug transporters such as OATP1B1 and OATP1B3 were involved. TM‐25659 had a significantly greater basal to apical transport rate (5.9‐fold) than apical to basal transport rate in the Caco‐2 cell monolayer, suggesting the involvement of an efflux transport system. Further studies using inhibitors of efflux transporters and overexpressing cells revealed that MRP2 was involved in the transport of TM‐25659. These results, taken together, suggested that TM‐25659 can be actively influxed into hepatocytes and undergo biliary excretion without substantial metabolism. Additionally, TM‐25659 inhibited the transport activities of OATP1B1 and OATP1B3 with IC₅₀ values of 36.3 and 25.9 μm , respectively. TM‐25659 (100 μm ) increased the accumulation of the probe substrate by 160% and 213%, respectively, through the inhibition of efflux function of P‐gp and MRP2. In conclusion, OATP1B1, OATP1B3, P‐gp and MRP2 might be major transporters responsible for the pharmacokinetics and drug–drug interaction of TM‐25659, although their contribution to in vivo pharmacokinetics needs to be further investigated. Copyright © 2013 John Wiley & Sons, Ltd.     

A Novel TAZ Gene Mutation and Mosaicism in a Polish Family with Barth Syndrome

Barth syndrome (BTHS) is an X‐linked recessive disease primarily affecting males. Clinically, the disease is characterized by hypertrophic or dilated cardiomyopathy, skeletal myopathy, chronic/cyclic neutropenia, 3‐methylglutaconic aciduria, growth retardation and respiratory chain dysfunction. It is caused by mutations in the TAZ gene coding for the tafazzin protein which is responsible for cardiolipin remodeling. In this work, we present a novel pathogenic TAZ mutation c.83T>A, p.Val28Glu, found in mosaic form in almost all female members of a Polish family. Sanger sequencing of DNA from peripheral blood and from epithelial cells showed female mosaicism in three generations. This appears to be a new mechanism of inheritance and further research is required in order to understand the mechanism of this mosaicism. We conclude that BTHS genetic testing should include two or more tissues for women that appear to be noncarriers when blood DNA is initially tested. The results of our study should not only be applicable to BTHS families, but also to families with other X‐linked diseases.     

帕比司他抑制卵巢癌裸鼠移植瘤上皮-间质转化和生长机制探讨

目的　探究帕比司他（LBH589）对卵巢癌裸鼠移植瘤上皮-间质转化（EMT）、生长及具有PDZ结合域的转录共刺激因子（TAZ）/表皮生长因子受体（EGFR）通路的影响。方法　将造模成功的雌性BALB/c裸鼠50只按随机数字表法分成5组（n=10）：阳性对照组（腹腔注射5 mg/kg贝伐单抗）、模型组（腹腔注射等量生理盐水）以及LBH589低、中、高（腹腔注射10、30及50 nmol/L LBH589）浓度组,干预后每3 d测量各组瘤体积,计算抑瘤率。干预14 d后,采用Western blot法检测各组瘤组织TAZ、EGFR、上皮钙黏附蛋白E（E-Cad）、波形蛋白（Vim）表达。结果　与模型组比较,3 d时阳性对照组、LBH589高浓度组小鼠皮下移植瘤体积减小（P＜0.05）,6 d、9 d、12 d时LBH589各浓度组、阳性对照组小鼠皮下移植瘤体积均减小（P＜0.05）;与阳性对照组比较,LBH589低、中浓度组小鼠9 d、12 d时皮下移植瘤体积增大,12 d时抑瘤率降低（P＜0.05）;与LBH589低、中浓度组相比,干预9 d、12 d LBH589高浓度组小鼠皮下移植瘤体积减小,12 d LBH589高浓度组抑瘤率升高（P＜0.05）;与模型组比较,阳性对照组、LBH589各浓度组移植瘤组织中E-Cad蛋白表达升高,Vim、TAZ、EGFR蛋白表达降低（P＜0.05）;与阳性对照组比较,LBH589低、中浓度组移植瘤组织中E-Cad蛋白表达降低,Vim、TAZ、EGFR蛋白表达升高（P＜0.05）;与LBH589低、中浓度组相比,LBH589高浓度组移植瘤组织中E-Cad蛋白表达升高,Vim、TAZ、EGFR蛋白表达降低（P＜0.05）。结论　TAZ/EGFR通路在卵巢癌裸鼠移植瘤生长和EMT中有重要作用,LBH589可能通过抑制TAZ/EGFR通路激活,发挥抑制卵巢癌裸鼠移植瘤生长和EMT的作用。