Tumor Cell-Derived Extracellular Vesicles Promote the Growth, Metastasis and Chemoresistance in Cholangiocarcinoma by Delivering microRNA-210 to Downregulate RECK.

The development of cholangiocarcinoma (CCA) can be regulated by extracellular vesicles (EVs). In this study, we intend to investigate whether tumor cell-derived EVs delivering microRNA (miR)-210 affect CCA development, involved with reversion-inducing-cysteine-rich protein with kazal motifs (RECK). In silico analysis was performed for identifying differentially expressed miRs and the downstream target genes. The CCA related microarray GSE77984 was used to verify the expression of the target genes in CCA tissue samples. Targeting relationship between miR-210 and RECK was assayed. EVs were extracted from CCA cells, followed by co-culture with CCA cells. The in vitro and in vivo roles of tumor cell-derived EVs on the growth and metastasis of CCA cells were assayed. Upregulated miR-210 and downregulated RECK were found in CCA. CCA cells could uptake tumor cell-derived EVs, and the EVs could promote their migration, invasion, and chemoresistance. RECK expression could be target and inhibited by miR-210. It was further confirmed in vivo that miR-210 shuttled by tumor cell-derived EVs could specifically inhibit RECK expression, which promotes growth, metastasis and chemoresistance of CCA cells. Our current study highlighted that tumor cell-derived EVs could deliver miR-210 to CCA cells, where miR-210 specifically decreases RECK expression, which facilitates growth, metastasis and chemoresistance in CCA.

[Retracted] miR­494 promotes cell proliferation, migration and invasion, and increased sorafenib resistance in hepatocellular carcinoma by targeting PTEN.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that the western blotting data in Figs. 3 and 6 were strikingly similar to data appearing in different form in other articles by different authors at different research institutes. Owing to the fact that the contentious data in the above article were already under consideration for publication, or had already been published, elsewhere prior to its submission to Oncology Reports, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they agreed with the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [the original article was published in Oncology Reports 34: 1003­1010, 2015; DOI: 10.3892/or.2015.4030].

Sarcomatoid intrahepatic cholangiocarcinoma in a patient with poor prognosis: a case report and literature review.

Sarcomatoid intrahepatic cholangiocarcinoma (S-iCCA) is a rare histological variant of intrahepatic cholangiocarcinoma (iCCA). The diagnosis of S-iCCA is based on histopathological and immunohistochemical examinations, and S-iCCA often has a poorer prognosis than that of ordinary iCCA. In this article, we present the case of a 64-year-old man with S-iCCA who presented with intermittent right upper abdominal pain. The aim of this case report and literature review is to strengthen the understanding of S-iCCA among clinicians and reduce the incidence of missed clinical diagnoses.

WITHDRAWN: Novel long-acting BF-30 conjugate corrects pancreatic carcinoma via cytoplasmic membrane permeabilization and DNA-binding in tumor-bearing mice.

This article has been withdrawn at the request of the editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/our-business/policies/article-withdrawal.

Downregulation of CRABP2 Inhibit the Tumorigenesis of Hepatocellular Carcinoma In Vivo and In Vitro.

Cellular retinoic acid-binding protein 2 (CRABP2) binds retinoic acid (RA) in the cytoplasm and transports it into the nucleus, allowing for the regulation of specific downstream signal pathway. Abnormal expression of CRABP2 has been detected in the development of several tumors. However, the role of CRABP2 in hepatocellular carcinoma (HCC) has never been revealed. The current study aimed to investigate the role of CRABP2 in HCC and illuminate the potential molecular mechanisms. The expression of CRABP2 in HCC tissues and cell lines was detected by western blotting and immunohistochemistry assays. Our results demonstrated that the expression levels of CRABP2 in HCC tissues were elevated with the tumor stage development, and it was also elevated in HCC cell lines. To evaluate the function of CRABP2, shRNA-knockdown strategy was used in HCC cells. Cell proliferation, metastasis, and apoptosis were analyzed by CCK-8, EdU staining, transwell, and flow cytometry assays, respectively. Based on our results, knockdown of CRABP2 by shRNA resulted in the inhibition of tumor proliferation, migration, and invasion in vitro, followed by increased tumor apoptosis-related protein expression and decreased ERK/VEGF pathway-related proteins expression. CRABP2 silencing in HCC cells also resulted in the failure to develop tumors in vivo. These results provide important insights into the role of CRABP2 in the development and development of HCC. Based on our findings, CRABP2 may be used as a novel diagnostic biomarker, and regulation of CRABP2 in HCC may provide a potential molecular target for the therapy of HCC.

Therapeutic Effects of Paclitaxel Loaded Polyethylene Glycol-Polylactic Acid-Glycolic Acid Copolymer Nanoparticles on Pancreatic Cancer in Rats.

To establish a simple and safe method for the preparation of paclitaxel PEG-PLGA nanoparticles emulsified in tpgs (PTX-pegpllga-np), for high drug loading; and to study its effect on proliferation and apoptosis of human pancreatic cancer cell line MIAPACA-2. PTX-PEG-PLGA-NP was prepared by one-step precipitation, using tpgs as emulsifier. The drug loading and particle size were used as an index to optimize the formulation, and the physical and chemical properties such as in vitro release and stability were characterized. The uptake of fluorescein coumarin 6 (C6) loaded PEG-PLGA-NP by MIAPACA-2 cells was observed by fluorescence microscope, and the growth and apoptosis of MIAPACA-2 cells after PTX-PEG-PLGA-NP were detected by MTT and flow cytometry respectively. The entrapment efficiency of the nanoparticles was 90.26%, the drug loading was 10.13%, the average particle size was 92.3±3.1 nm, and the zeta potential was 10.48±1.54 mV. The cumulative releases of nano preparation and general preparation (Taxol injection) in four hours were 25.9% and 98.5%, respectively; and the former had a strong sustained-release effect. The results of cell uptake experiments showed that the uptake of c6-PEG-PLGA-NP by MIAPACA-2 cells increased gradually with time. MTT results showed that PTX-PEG-PLGA-NP had no significant difference in the inhibition rate of MIAPACA-2 cells compared with PTX group. Flow cytometry showed that PTX-PEG-PLGAnp was superior better than PTX in inducing apoptosis in MIAPACA-2 cells. The tpgs emulsification method is simple and environment-friendly. The paclitaxel loaded nanoparticles prepared through the optimization of the formulation have large drug loading capacity and uniform particle size, which can target the pancreatic cancer MIAPACA-2 cells, and do not weaken its ability to inhibit the growth of MIAPACA-2 cells. The nanoparticles also induce apoptosis in cancer MIAPACA-2 cells, and could be used for further clinical treatment of pancreatic cancer.

miR-330-3p suppresses liver cancer cell migration by targeting MAP2K1.

MicroRNAs, considered as a promising focus for the treatment of tumors, are key regulators of a large number of genes. The aim of the present study was to investigate the biological functions of microRNA (miR)-330-3p in liver cancer as it had been identified previously that miR-330-3p was deregulated in liver cancer. In order to identify the function of miR-330-3p in liver cancer, the expression of miR-330-3p was determined in liver cancer tissues and adjacent non-tumor tissues using reverse transcription-quantitative polymerase chain reaction analysis. To elucidate the function of miR-330-3p in liver cancer, miR-330-3p was overexpressed using mimic transfection. Cell migration was inhibited by miR-330-3p in liver cancer cells. The miRNA target prediction databases were used to identify potential target genes of miR-330-3p in liver cancer. The RNA level of mitogen-activated protein kinase kinase 1 (MAP2K1) was downregulated by miR-330-3p in liver cancer cells. In conclusion, miR-330-3p suppresses cell migration by targeting MAP2K1 in liver cancer cells.

LncRNA SNHG15 promotes hepatocellular carcinoma progression by sponging miR-141-3p.

Small nucleolar RNA host gene 15 (SNHG15) is a long noncoding RNA (lncRNA), which promotes progression of multiple cancers. Its specific function in hepatocellular carcinoma (HCC), however, is uncertain. The aims of our study were, therefore, to explore the role of SNHG15 in HCC. SNHG15 and miR-141-3p expression were assessed via quantitative real-time PCR (qRT-PCR) in 58 paired HCC samples and adjacent matched adjacent normal tissues. CCK-8 assay, flow cytometric examination, and wound healing/invasion assays were used to respectively assess how SNHG15 influences cell proliferation, the cell cycle, and the migratory and invasive potential of HCC cells. MicroRNA (miRNAs) that targeted SNHG15 was screened by Starbase2.0 and identified by RNA immunoprecipitation and luciferase reporter assays. SNHG15 expression was markedly increased, whereas miR-141-3p expression was substantially reduced in HCC cells and tissue samples relative to normal controls. When SNHG15 was knocked down, this resulted in a significant disruption to the proliferation, as well as the invasive and migratory ability of these HCC cells. miR-141-3p was also found to be an SNHG15 target in HCC cells. Furthermore, miR-141-3p inhibitor partially reversed the observed SNHG15 depletion-mediated reduction in HCC proliferation, migration, and invasion. By repressing miR-141-3p, SNHG15 could modulate zinc finger E-box binding homeobox 2 (ZEB2) and E2F transcription factor 3 (E2F3) expression, both of which are miR-141-3p targets. These finding suggested that SNHG15 promoted HCC progression via negative regulation of miR-141-3p, thus identifying a potential novel HCC treatment pathway.

Retracted: MicroRNA-34a inhibit hepatocellular carcinoma progression by repressing hexokinase-1.

Hepatocellular carcinoma (HCC) is known as a frequent type of primary cancer in the liver, and it is the third-most common cause of cancer-related death all over the world. However, the molecular mechanism in the progression of HCC is still unclear. The current study was designed to investigate the expression and function of microRNA-34a (miR-34a) in HCC. In HCC tissues and cells, the expression levels of miR-34a were analyzed by quantitative real-time polymerase chain reaction. The association between the level of miR-34a and hexokinase (HK)-1 was also investigated via luciferase reporter assay. Cell viability and proliferation were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry. To assess whether miR-34a can limit tumor growth in vivo, animal models and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were used for examining the role of miR-34a on the development of HCC and cell apoptosis. The expression level of miR-34a was reduced in HCC samples and cells. The expression of miR-34a was associated with the viability and proliferation capacity of HCC cells, and miR-34a could inhibit HCC cells proliferation by inhibiting HK1. In the mouse model of HCC, volumes and weight of the tumors were significantly decreased by transfection with miR-34a mimic compared with the control group. Furthermore, miR-34a mimics could induce apoptosis in a greater proportion of cells compared with the control group. Taken together, the data may provide some novel insights into the molecular mechanism of miR-34a and HK1 in the progression of HCC. Thus, miR-34a/HK1 axis might be a novel promising therapeutic target for treating HCC.

Percutaneous Transhepatic Gallbladder Drainage Combined with Laparoscopic Cholecystectomy versus Emergency Laparoscopic Cholecystectomy in Acute Complicated Cholecystitis: Comparison of Curative Efficacy.

Controversy exists on the suitability of laparoscopic cholecystectomy (LC) in acute cholecystitis, especially in patients with severe comorbidities. Recently, many nonsurgical departments have indicated a preference for percutaneous transhepatic gallbladder drainage (PTGBD), but surgeons consider LC as the final treatment option for cholecystitis. This analysis evaluated the curative efficacy of PTGBD in combination with LC as compared with emergency LC (e-LC). We retrospectively analyzed clinical data of 86 patients with acute complicated cholecystitis. Patients were divided into two groups as those who received e-LC and those who underwent PTGBD combined with LC (PTGBD+LC), and baseline characteristics, perioperative data, and operative parameters were compared to check for intergroup differences. Baseline characteristics were similar for the study groups. However, although the operating duration (P = 0.12) and postoperative hospital stay (P = 0.39) did not evidence significant differences, the PTGBD+LC group had significantly better outcomes than the e-LC group with regard to blood loss (P < 0.05), peritoneal drainage duration (P < 0.05), and time to postoperative resumption of oral intake (P < 0.05). Moreover, conversion to open surgery, complications during LC, and mortality rate were all higher in the e-LC group. PTGBD combined with LC is an effective treatment for acute complicated cholecystitis, especially in elderly patients or those with serious comorbidities. To some extent, the curative effect of this method can be considered superior to that of emergency LC.

Evaluation of the Safety and Efficacy of Percutaneous Transhepatic Gallbladder Drainage Combined with Laparoscopic Cholecystectomy for Treating Acute Complicated Cholecystitis.

The rate of acute cholecystitis in patients with severe underlying diseases is currently increasing. Several studies have reported percutaneous transhepatic gallbladder drainage (PTGBD) combined with laparoscopic cholecystectomy (LC) as a safe and reliable therapeutic option in such patients. This study aimed to elucidate the optimal time interval between PTGBD and LC. In total, 65 patients with acute complicated cholecystitis from our hospital were divided into two groups, short-term LC (sLC) and postponed LC (pLC) group according to whether the procedure was performed within 5 days of gallbladder drainage or after 5 days, respectively. The complications after PTGBD, rate of conversion to open surgery, and complications and mortality after LC were compared between the groups. The sLC group showed significantly lesser operating time, blood loss, postoperative peritoneal drainage time, postoperative oral intake time, and complications compared to the pLC group (P < 0.05). Other factors such as the length of hospital stay (LOS), conversion to open cholecystectomy, and mortality were not statistically significant between the groups. Combined treatment with PTGBC and sLC showed superior outcomes compared to PTGBC and pLC for acute cholecystitis in severely ill patients, thus constituting a feasible and secure treatment option in specialized centers.

Ligands of the peroxisome proliferator-activated receptor γ inhibit hepatoce llular carcinoma cell proliferation.

This study was designed to investigate the regulatory role of the peroxisome proliferator-activated receptor γ (PPARγ) in the growth of hepatocellular carcinoma cells under the hypothesis that the levels of the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) mRNA and the phosphorylated Akt (pAkt) protein would be affected by the presence of different receptor ligand concentrations. SMMC-7721 hepatocellular carcinoma cells were cultured in the presence of different concentrations of either 15-deoxyprostaglandin J2 (15-d-PGJ2) or pioglitazone and experiments were conducted in order to determine cell growth changes and measure levels of PTEN mRNA and pAkt protein. Our results after treatment with MTT showed the addition of ligands to the cultured cells inhibited their proliferation in a time- and dose-dependent manner. Also, flow cytometry after PI treatment showed the presence of ligands in the growth media could increase the proportion of G0/G1 phase cells, and decrease the proportion of S phase cells. Finally, the same cells exhibited increased levels of the PTEN mRNA by RT-PCR and pAkt protein by western blot analysis. Taken together, our results support the notion that PPARγ ligands can inhibit the proliferation of hepatocellular carcinoma cells in a time- and dose-dependent manner, and that this is at least in part due to the resulting upregulation of PTEN expression.

Duel-stage treatment for biliary cysts with cholangitis during pregnancy.

BACKGROUND & OBJECTIVE: Biliary cysts in pregnant women are a complex medical issue, especially when complicated with cholangitis. It is a serious and life-threatening diagnosis that can seriously endanger both the expectant mother and the fetus. However, during pregnancy, surgical treatment would lead to further complications and higher fetal mortality. Here, we propose a novel therapeutic approach that would be safe for both mother and child during pregnancy, with a definitive treatment postponed until after delivery. METHODS: In this retrospective study we have summarized the clinical course of six adult patients diagnosed with choledochal cysts during pregnancy. Treatment was conducted in two stages, firstly, percutaneous cholecystostomy under ultrasound guidance and sustained negative pressure suction until delivery, and secondly, selective choledochal cyst excision when the patients recovered from delivery. RESULTS: All the six patients gave birth to healthy babies. Four patients had Type-I choledochal cysts, and underwent Roux-en-Y hepaticojejunostomy surgery. Two patients had a Type-IV choledochal cyst. The first patient with Type-IV choledochal cyst underwent anastomosis between the secondary hepatic bile duct and jejunum and the second patient underwent laparoscopic cyst internal drainage. No serious complications were recorded after gallbladder drainage or during the perioperative period. CONCLUSIONS: Based on our single-centre experience we can conclude that treatment of choledochal cyst with cholangitis during pregnancy can be conducted safely and efficiently through the two stages strategy that we proposed in this paper. The first stage should be percutaneous cholecystostomy followed by elective surgical treatment following delivery.

MiR-892a Promotes Hepatocellular Carcinoma Cells Proliferation and Invasion Through Targeting CD226.

Our study is aim to investigate the influence of miR-892a on proliferative and invasive activities of human hepatocellular carcinoma (HCC) cells through regulating CD226 expression. QRT-PCR was used to detect the expression levels of miR-892a and CD226 mRNA in HCC tissues and adjacent tissues or HCC cells and normal cells whereas Western Blot was used to detect the CD226 protein expression in tissue and cell samples. Then HuH-7 cell line was selected for following assays and respectively transfected with miR-892a mimics, miR-NC, Plenti-GIII-Ubc-CD226, and Plenti-GIII-Ubc followed by qRT-PCR assay to detect the miR-892a and CD226 expression. The luciferase reporter assay was conducted to determine if miR-892a directly targeted CD226 and then CCK-8 assay, wound healing assay, Transwell assay, and flow cytometry were used to detect cell proliferation, migration, invasion ability, cell cycle, and cell apoptosis. What's more, relationships between expression levels of miR-892a or CD226 and overall survival (OS) or disease-free survival (DFS) of HCC patients were investigated based on TCGA database. MiR-892a was high-expressed in HCC tissues or cells while CD226 was low-expressed. MiR-892a directly targeted CD226 and up-regulating miR-892a expression could promote proliferative, migrating, and invasive activities of HCC cells. Different expression levels of miR-892a and CD226 both related to the prognosis of HCC. MiR-892a promotes hepatocellular carcinoma cells proliferation and invasion through regulating CD226 expression. J. Cell. Biochem. 118: 1489-1496, 2017. © 2016 Wiley Periodicals, Inc.

miR-494 promotes cell proliferation, migration and invasion, and increased sorafenib resistance in hepatocellular carcinoma by targeting PTEN.

MicroRNA-494 (miR-494) acts as an oncomiR and is involved in tumor development, progression and metastasis, and confers resistance to chemotherapeutic drugs by targeting a number of molecules in several human cancers. However, the function and underlying molecular mechanism of miR-494 in hepatocellular carcinoma (HCC) has not been totally elucidated. In the present study, we determined the role played by miR-494 in HCC tissues and HCC cell lines using quantitative RT-PCR (RT-qPCR). The results showed that, miR-494 was significantly upregulated in HCC tissues and HCC cell lines. Additionally, a high miR-494 expression positively correlated with tumor differentiation (P<0.01), TNM stage (P<0.01) and lymph node metastasis (P<0.01). Luciferase reporter assays confirmed that miR-494 binds to the 3'-untranslated region (3'-UTR) of the phosphatase and tensin homolog (PTEN) mRNA and represses its translation. Functional analyses indicated that the upregulation of miR-494 promoted cell viability, migration and invasion, decreased cell apoptosis and cell cycle arrest at G1 stage, and conferred sorafenib resistance to HCC cell lines. Underexpression of PTEN by siRNA significantly attenuated the inhibitory effects of anti-miR-494 on the proliferation, migration and invasion of liver cancer cells. Mechanistic investigations revealed that miR-494 suppressed the expression of PTEN but increased the expression of PI3K and p-Akt, which contribute to the promotion of proliferation, migration and invasion, and increased sorafenib resistance to HCC cell lines. These findings suggested that miR-494 is a potential candidate for HCC therapeutics.