Brachyury promotes proliferation and migration of hepatocellular carcinoma via facilitating the transcription of NCAPG2.

Hepatocellular carcinoma (HCC) has a poor prognosis because of its limited drug responses in clinical trials. Therefore, it is crucial to clarify the molecular mechanisms of HCC progression to identify new diagnostic markers and therapeutic targets. Here, we report that brachyury, which regulates the gene encoding the non-SMC condensin II complex subunit G2 (NCAPG2), promotes tumorigenesis in HCC. Knockdown of brachyury led to inhibition of cancer progression in vitro and in vivo. Chromatin immunoprecipitation-sequencing data indicated that the oncogene NCAPG2 is a direct target of brachyury. Furthermore, NCAPG2 knockdown inhibited the proliferation and migration of HCC cells and attenuated brachyury-induced tumorigenesis. Overexpression and decreased DNA methylation of NCAPG2 were associated with a poor prognosis, and NCAPG2 was positively correlated with various immune cell infiltrates, cancer-associated fibroblasts, and immune checkpoint molecule expression levels in the tumor microenvironment. Moreover, the effectiveness of immune checkpoint blockade was decreased in the high NCAPG2 expression group. Together, these findings demonstrated a coregulatory effect of the brachyury/NCAPG2 axis during HCC progression.

Long Non-coding RNA RP11-395G23.3 Acts as a Competing Endogenous RNA of miR-124-3p to Regulate ROR1 in Anaplastic Thyroid Carcinoma.

Anaplastic thyroid carcinoma (ATC) is one of the most aggressive human malignancies with poor prognosis. However, the underlying mechanisms of ATC remain to be elucidated. Recently, increasing studies have focused on competitive endogenous RNA (ceRNA) to discover valuable biomarkers for the diagnosis of ATC. The present study identified 705 differentially expressed mRNAs and 47 differentially expressed lncRNAs. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were also conducted. Additionally, an lncRNA/miRNA/mRNA network was constructed which included 1103 regulatory relations. The upregulation of RP11-395G23.3 in ATC cells was confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). In the loss of function assays, results suggested silencing of RP11-395G23.3 inhibited cell proliferation and induced cell apoptosis. Mechanically, RP11-395G23.3 could increase ROR1 via sponging miR-124-3p as a ceRNA. Moreover, ROR1 expression was decreased with the downregulation of RP11-395G23.3, but was rescued by the co-transfection of the miR-124-3p inhibitor in ATC cells. Our research suggested that the RP11-395G23.3/miR-124-3p/ROR1 axis potentially acted as a potential target for the diagnosis of ATC.

Papillary Thyroid Microcarcinoma: A Nomogram Based on Clinical and Ultrasound Features to Improve the Prediction of Lymph Node Metastases in the Central Compartment.

Background: Accurate preoperative identification of central lymph node metastasis (CLNM) is essential for surgical protocol establishment for patients with papillary thyroid microcarcinoma (PTMC). We aimed to develop a clinical and ultrasound characteristics-based nomogram for predicting CLNM. Methods: Our study included 399 patients who were pathologically diagnosed with PTMC between January 2011 and June 2018. Clinical and ultrasound features were collected for univariate and multivariate analyses to determine risk factors of CLNM. A nomogram comprising the prognostic model to predict the CLNM was established, and internal validation in the cohort was performed. The Cox regression model was used to determine the risk factors for recurrence-free survival (RFS) and cumulative hazard was calculated to predict prognosis. Results: Three variables of clinical and US features as potential predictors including sex (odd ratio [OR] = 1.888, 95% confidence interval [CI], 1.160-3.075; P =0.011), tumor size (OR = 1.933, 95% CI, 1.250-2.990; P =0.003) and ETE (OR = 6.829, 95% CI, 3.250-14.350; P <0.001) were taken into account. The predictive nomogram was established by involving all the factors above used for preoperative prediction of CLNM in patients with PTMC. The nomogram showed excellent calibration in predicting CLNM, with area under curves (AUC) of 0.684 (95% CI, 0.635 to 0.774). Furthermore, tumor size, multifocality, presence of ETE, vascular invasion, and CLNM were the significant factors related to the RFS. Conclusion: Through this easy-to-use nomogram by combining clinical and US risk factor, the possibility of CLNM can be objectively quantified preoperatively. This prediction model may serve as a useful clinical tool to help clinicians determine an individual's risk of CLNM in PTMC, thus make individualized treatment plans accordingly.

Management of Clinically Solitary Papillary Thyroid Carcinoma Patients According to Risk-Scoring Model for Contralateral Occult Carcinoma.

Objective: The aim of this study was to investigate risk factors of occult carcinoma in clinically solitary papillary thyroid carcinoma (PTC) patients, and to put emphasis on the predictive value of risk-scoring model to determine the optimal scope of surgery. Methods: A total of 573 clinically solitary PTC patients who underwent total thyroidectomy (TT) from two hospitals were retrospectively analyzed. Clinicopathological features were collected, univariate and multivariate analyses were performed to determine risk factors of occult carcinoma. The Cox proportional hazards model was used to analyze the risk factors of recurrence. A scoring model was constructed according to independent risk factors of contralateral occult carcinoma. Results: 19.2% of clinically solitary PTC patients had occult carcinoma, among which 3.7% patients had ipsilateral occult carcinoma and 15.5% patients had contralateral occult carcinoma. Factors such as male, the presence of benign nodule, and vascular invasion increase the risk of ipsilateral occult carcinoma. Tumor size >1 cm, the presence of benign nodule, extrathyroidal extension, central lymph node metastasis, lateral lymph node metastasis are independent predictors of contralateral occult carcinoma. Contralateral occult carcinoma is the independent predictor of recurrence. A 10-point risk-scoring model was established to predict the contralateral occult carcinoma in clinically solitary PTC patients. Conclusion: Lobectomy is sufficient for clinically solitary PTC patients with risk factors of ipsilateral occult carcinoma. For clinically solitary PTC patients with score ≥4, careful preoperative evaluations are required to rule out the contralateral occult carcinoma. Even if contralateral occult carcinoma is not detected preoperatively, TT is recommended for high-risk patients.

The therapeutic effect of the BRD4-degrading PROTAC A1874 in human colon cancer cells.

A1874 is a novel BRD4-degrading proteolysis targeting chimera (PROTAC). In primary colon cancer cells and established HCT116 cells, A1874 potently inhibited cell viability, proliferation, cell cycle progression, as well as cell migration and invasion. The BRD4-degrading PROTAC was able to induce caspase and apoptosis activation in colon cancer cells. Furthermore, A1874-induced degradation of BRD4 protein and downregulated BRD-dependent genes (c-Myc, Bcl-2, and cyclin D1) in colon cancer cells. Significantly, A1874-induced anti-colon cancer cell activity was more potent than the known BRD4 inhibitors (JQ1, CPI203, and I-BET151). In BRD4-knockout colon cancer cells A1874 remained cytotoxic, indicating the existence of BRD4-independent mechanisms. In addition to BRD4 degradation, A1874 cytotoxicity in colon cancer cells was also associated with p53 protein stabilization and reactive oxygen species production. Importantly, the antioxidant N-acetyl-cysteine and the p53 inhibitor pifithrin-α attenuated A1874-induced cell death and apoptosis in colon cancer cells. In vivo, A1874 oral administration potently inhibited colon cancer xenograft growth in severe combined immuno-deficient mice. BRD4 degradation and p53 protein elevation, as well as apoptosis induction and oxidative stress were detected in A1874-treated colon cancer tissues. Together, A1874 inhibits colon cancer cell growth through both BRD4-dependent and -independent mechanisms.

Modafinil ameliorated pancreatic injury and inflammation through upregulating SNIP1.

Acute pancreatitis (AP) is the inflammatory response of the exocrine pancreas to various causes. Modafinil has significant anti-inflammation and anti-oxidation effects. No experiment has assessed the effects of modafinil on AP. Thus, the study aims to study the effects of modafinil on AP and its potential mechanism in vivo and vitro. 5% sodium taurocholate was retrograde injected into pancreatic duct to establish AP rat model. The severity of AP was detected by HE staining, serum amylase and lipase levels. The inflammation, oxidative stress and apoptosis were detected separately by ELISA, MDA and SOD kits, tunnel staining and Western blotting in rats. Besides, SNIP1 expression was analyzed by qPCR and Western blotting. In vivo, AR42J cells were stimulated by cerulein and lipopolysaccharide to establish AP cell model. Flow cytometry examined cell apoptosis. After the plasmids silencing SNIP1 were transfected into AP cells, the inhibitory effects of modafinil on inflammation, oxidative stress and apoptosis were significantly reversed. The results indicated that modafinil showed significant curative and therapeutic effects by regulating SNIP1 level.

Influence of Tumor Number on Clinicopathologic Features and Outcomes of Patients With Papillary Thyroid Carcinoma.

OBJECTIVES: The purpose of this study was to investigate the significance of tumor number on clinicopathologic factors and outcomes of patients with papillary thyroid carcinoma (PTC). METHODS: We retrospectively analyzed 667 patients with PTC. We compared clinicopathologic features of patients with a different tumor number. Cox proportional hazards model was used to analyze risk factors of recurrence. RESULTS: In papillary thyroid microcarcinoma (PTMC), the increase in the number of tumor foci was related to a higher risk of minimal extrathyroidal extension (ETE) and lymphovascular invasion (P < .05). Patients with PTMC with four or more foci had a significantly higher risk of central lymph node metastasis (CLNM) and lateral lymph node metastasis (LLNM) than patients with solitary tumors (P < .05). Patients with macro-PTC with four or more foci and with three foci had a higher risk of gross ETE and lymphovascular invasion than patients with solitary tumors (P < .05). The increase in the tumor number was related to a higher risk of CLNM in macro-PTC (P < .05). The number of foci was the independent predictor of recurrence in patients with macro-PTC (P < .05). CONCLUSIONS: An increase in the number of tumors was associated with an increased risk of aggressive clinicopathologic features in PTMC and macro-PTC. The number of tumor foci could influence risk of recurrence in macro-PTC.

Significance of multifocality in papillary thyroid carcinoma.

BACKGROUND: Although multifocality is often observed in papillary thyroid carcinoma (PTC), the associations with clinicopathologic factors and the prognostic value are still controversial. We aimed to identify the risk factors for multifocality and bilaterality, and investigate the significance of multifocality on prognosis in all PTC, papillary thyroid microcarcinoma (PTMC) and non-PTMC. METHODS: Data from 635 patients who underwent lobectomy/total thyroidectomy plus cervical lymph node dissection for PTC were retrospectively analyzed. Clinicopathological factors associated with multifocal PTC and bilateral PTC were investigated by univariate analysis. Multivariate Cox regression analyses was used to identify the clinicopathological prognostic factors for recurrence-free survival. RESULTS: Multifocal and bilateral PTC were observed in 157 (24.7%) and 99 (15.6%) patients, respectively. The frequency of large diameter (>1.0 cm), extrathyroidal extension (ETE), vascular invasion and central lymph node metastases (CLNM) was higher in multifocal PTC than that of solitary PTC. Moreover, ETE, vascular invasion, CLNM and lateral lymph node metastasis (LLNM) were more frequent in patients with 3 or more tumor foci compared to those with 2 tumor foci and 1 tumor lesion. Bilateral PTC had higher rates of ETE and CLNM. Multifocality was found to be the predictor of recurrence in all PTC, PTMC, and non-PTMC. In addition, the risk of recurrence increased with increasing number of tumor foci in all multifocal PTC patients and multifocal non-PTMC patients. CONCLUSION: Although multifocality and bilaterality had more aggressive features in PTC, only multifocality was associated with the increased risk of recurrence. An increase in the number of tumors was associated with an increased risk of ETE, vascular invasion, CLNM and LLNM. The prognostic value of multifocality is particularly significant in non-PTMC patients.

Sinensetin flavone exhibits potent anticancer activity against drug-resistant human gallbladder adenocarcinoma cells by targeting PTEN/PI3K/AKT signalling pathway, induces cellular apoptosis and inhibits cell migration and invasion.

PURPOSE: The main focus of the current research work was to unveil the anticancer activity of the naturally occurring Sinensetin flavone against aggressive gall bladder cancer adenocarcinoma (GBAC) TJ-GBC2 cell line. Its effect of inducing apoptosis mediated via targeting PTEN/PI3K/AKT signalling pathway were also examined along with cell migration and invasion. METHODS: Cell proliferation was tested by MTT cell viability assay. Fluorescence microscopy was utilized to carry out apoptosis related studies via DAPI staining along with flow cytometry using annexin V/propidium iodide (PI) assay. Further, western blotting analysis was carried out to examine the effects of Sinensetin on the expressions of apoptosis-related proteins and Bax Bcl-2 along with PTEN/PI3K/AKT signalling pathway. The impact of the test molecule on cell migration and invasion was studied through wound healing assay and transwell cell invasion assay respectively. RESULTS: The results showed that Sinensetin treatment caused a significant retardation in cell viability, in a dose-dependent fashion. DAPI staining assay and annexin V/PI assay revealed that the cell viability of GBC cells was retarded due to induction of apoptosis. It was also associated with downregulation of Bcl-2 and upregulation of Bax levels. Further, wound healing assay and transwell cell invasion assay revealed that cell migration as well as cell invasion of cancer gallbladder cells was decreased in a concentration-dependent fashion. It was further seen that Sinensetin treatment resulted in inhibition of matrix metalloproteinase (MMP)-2 and enhancement of MMP-9 protein expressions. Results also showed that the tested molecule had the potential to inhibit PTEN/PI3K/AKT signalling pathway. CONCLUSION: In conclusion, the current study indicated that Sinensetin flavone has the potential to be developed as a candidate drug against gallbladder adenocarcinoma provided more toxicological and in vivo studies are carried out.

Axin1 inhibits proliferation, invasion, migration and EMT of hepatocellular carcinoma by targeting miR-650.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors with a high mortality rate and low survival rate. This study was designed to explore a novel molecular with high sensitivity and specificity, which can be applied in early diagnosis and therapeutic evaluation of HCC. The current study aims to investigate the effect and important role of Axin1 on cell proliferation, invasion, migration and epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma. qRT-PCR results showed lower Axin1 expression level and higher miR-650 expression level in HCC. Luciferase reporter assay was carried out to verify the negative correlation between Axin1 and miR-650 mRNA levels. CCK-8 assay results showed that the cell proliferation ability was significantly suppressed by Axin1 overexpression in SK-HEP-1 cells. The results in wound healing assay uncovered that cell migration ability was markedly suppressed by Axin1 overexpression. The results in trans-well invasion assay showed that Axin1 overexpression caused decreased invasive ability in SK-HEP-1 cells. The WB results showed that the protein level of E-cad was significantly increased and the protein levels of N-cad, vimentin and snail were obviously reduced following Axin1 overexpression. Whereas, the suppressive effects on cell proliferation, migration, invasion and EMT caused by Axin1 overexpression were abolished by miR-650 mimic. All the results in the current study confirmed the truth that Axin1 overexpression could suppress cell proliferation, migration, invasion and EMT by downregulating miR-650 expression.

Role of miR-18a and miR-25 disruption and its mechanistic pattern in progression of liver cancer.

This study examined the molecular mechanisms underlying the roles of the microRNAs miR-18a and miR-25 in the progression of human liver cancer. Liver cancer biopsies obtained from early-stage liver cancer patients were examined by qRT-PCR and Northern blotting to examine the expression of miR-18a and miR-25. Both microRNAs were overexpressed in mouse primary hepatocytes following transfection of the cells with vectors encoding the microRNAs. An analysis of biopsy samples from liver cancer patients indicated that both miR-18a and miR-25 were overexpressed during the early stages of liver cancer. Further, qRT-PCR and Northern blotting confirmed that both of these microRNAs play crucial roles in the progression of liver cancer. Our findings clearly indicate that miR-18a and miR-25 can be used as prognostic biomarkers for early-stage liver cancer. Hence, miR-18a and miR-25 may have value as prognostic indicators and may facilitate the development of novel therapeutics for liver cancer.

Predictive Factors for Lateral Lymph Node Metastasis and Skip Metastasis in Papillary Thyroid Carcinoma.

In papillary thyroid cancer (PTC) patients, cervical lymph node metastases are common, which disseminate sequentially from the central neck to the lateral neck. However, there is also a chaotic pattern of lymph node metastasis occasionally. In this study, we summarized 653 PTC patients who underwent thyroidectomy and central lymph node dissection with or without lateral lymph node dissection from two hospitals to investigate the pattern and risk factors of lateral lymph node metastasis (LLNM) and skip metastasis. LLNM was significantly associated tumor size > 1 cm, presence of extrathyroidal extension, tumors in the upper-lateral pole, and the number of metastatic lymph nodes in the central compartment. The frequency of skip metastasis was 22.5% (20 of 89 patients). Multivariate analyses showed tumor size ≤ 1 cm, and tumors in the upper-lateral pole were separately and independently associated with the risk of skip metastasis. Presence of LLNM affected the recurrence-free survival (RFS). RFS did not show the significantly difference between patients with LLNM and skip metastasis. Despite the low incidence of skip metastasis, attention should be paid to the possibility of LLNM even in the absence of central lymph node metastases. Besides, for patients with risk factors of LLNM or skip metastasis, detailed preoperative examination for the lateral compartment, especially the level III, is essential.

miR-1470 regulates cell proliferation and apoptosis by targeting ALX4 in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. MicroRNAs (miRNAs) have been proven to play essential roles in different cancers, including HCC. The current study was mainly focused on the role of miR-1470 in HCC progression. METHODS: Quantitative real-time PCR (qRT-PCR) was performed to detect the expression levels of miR-1470 and Aristaless-like homeobox-4 (ALX4). The CCK-8 and EdU assays were used to examine cell proliferation. Flow cytometric analysis was used to elucidate the cell cycle and cell apoptosis. A xenograft tumor assay was carried out to verify the effect of miR-1470 on tumor formation in vivo. RESULTS: According to the qRT-PCR assay, miR-1470 was proven to be overexpressed in HCC. As shown by the CCK-8 assay, EdU assay and flow cytometric analysis, miR-1470 overexpression promoted cell proliferation and inhibited cell apoptosis. ALX4 was proven via a dual luciferase reporter assay to be a downstream target gene of miR-1470. ALX4 was downregulated in HCC. The results of a rescue assay revealed that miR-1470 had an oncogenic role in HCC by regulating ALX4. CONCLUSION: miR-1470 exhibits an oncogenic role in HCC by targeting ALX4. The data from our study may provide novel insight for the identification of new biomarkers and treatment strategies for HCC.

Comparison on the Efficacy of Three Duct Closure Methods after Laparoscopic Common Bile Duct Exploration for Choledocholithiasis.

BACKGROUND Laparoscopic common bile duct exploration (LCBDE) is currently the best approach for complex cases of choledocholithiasis or the cases of endoscopic retrograde cholangiopancreatography (ERCP) failure. Nevertheless, there is no clear consensus on the optimal duct closure method after LCBDE. The purpose of this study was to evaluate the efficacy of 3 duct closure methods after LCBDE for choledocholithiasis. MATERIAL AND METHODS In this analysis, 189 patients with choledocholithiasis underwent LCBDE between June 2014 and December 2018. According to different duct closure methods, these patients were divided into T-tube drainage (TTD) group (n=66), common suture group (n=64) and barbed suture group (n=59). The operation time, suturing time, amount of intraoperative bleeding, tube-carried time, length of stay (LOS), hospitalization costs, pre- and post-operative common bile duct (CBD) diameters were all compared among the 3 groups. Six months after discharge, the incidence of complications and recurrent stones was observed. RESULTS The operation time, suturing time, and amount of intraoperative bleeding in barbed suture group were both significantly less than those in the common suture group and the TTD group (P<0.01). When compared with the TTD group, the suturing time, tube-carried time, and LOS were decreased markedly in the common suture group and the barbed suture group (P<0.01). The post-operative CBD diameters in the 3 groups were all significantly larger than the pre-operative CBD diameters (P<0.01). There was no statistical significance among the 3 groups regarding the incidence of complications and recurrent stones (P>0.05). CONCLUSIONS Barbed suture shortened the suturing time, operation time, tube-carried time, and LOS, and lessened the amount of intraoperative bleeding in patients with choledocholithiasis after LCBDE. It was more effective than the common suture and TTD.

OCIAD1 promoted pancreatic ductal adenocarcinoma migration by regulating ATM.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive neoplastic disease, characterized with poor outcomes and a 5-year survival rate less than 5%. Dysregulation or dysfunction of immune response factors contribute to cancer development. In this study, we found that OCIAD1 is high expressed in pancreatic cancer gene chip, and verified OCIAD1 associating with cancer malignancy in specimens from patients with PDAC. OCIAD1 down-regulation inhibited PDAC cell lines migration and vice versa. Further analysis of pancreatic cancer gene chip found OCIAD1 high expression was associating with low ATM expression. Then we proved that OCIAD1 regulated ATM to affect the migration of PDAC. Thus we concluded that high OCIAD1 levels in PDAC promoted tumor cells migration. OCIAD1 exerted its effects by regulating ATM.

Dual PI3K-BRD4 Inhibitor SF1126 Inhibits Colorectal Cancer Cell Growth in Vitro and in Vivo.

BACKGROUND/AIMS: Bromodomain-containing protein 4 (BRD4) and phosphatidylinositol 3-kinase (PI3K) are key oncogenic cascades in colorectal cancer (CRC). SF1126 is a novel and potent PI3K-BRD4 dual inhibitor. METHODS: CRC cells and human colon epithelial cells were treated with SF1126. Cell survival was tested by MTT and soft agar colony formation assays. Cell proliferation was tested by BrdU ELISA method. Cell apoptosis was tested by a TUNEL staining method and Histone DNA ELISA. Western blotting was utilized to test the signaling proteins. A HT-29 xenograft mice model was established to study the anti-tumor activity of SF1126 in vivo. RESULTS: SF1126 potently inhibited the survival, proliferation, and progression of the cell cycle in an established CRC cell line (HT-29) and primary human colon cancer cells. Significant activation of apoptosis was detected in SF1126-treated CRC cells. In CRC cells, SF1126 blocked Akt-mammalian target of rapamycin (mTOR) complex1/2 signaling and downregulated BRD4 target proteins (Myc and cyclin D1). Further studies showed that SF1126 activated p38 signaling in CRC cells. In contrast, the p38 inhibitors or p38 short hairpin RNA inhibited SF1126-induced cytotoxicity and apoptosis in CRC cells. In vivo, subcutaneous administration of SF1126 significantly inhibited HT-29 xenograft tumor growth in nude mice. CONCLUSION: SF1126 inhibits CRC cell growth possibly by targeting PI3K-Akt-mTOR, BRD4, and p38 signaling.

MicroRNA-7 inhibits colorectal cancer cell proliferation, migration and invasion via TYRO3 and phosphoinositide 3-kinase/protein B kinase/mammalian target of rapamycin pathway suppression.

MicroRNA­7 (miR­7) is a non­coding RNA that inhibits colorectal cancer (CRC) cell proliferation, migration and invasion. miR­7 effectively silences TYRO3 expression, and the close association between TYRO3 and CRC has previously been reported. Therefore, the present study aimed to assess the roles and molecular mechanisms of TYRO3 and miR­7 in the development of CRC. The expression levels of miR­7 and TYRO3 in CRC tissues and cell lines were detected by reverse transcription­quantitative polymerase chain reaction. A dual­luciferase reporter assay was also performed to confirm whether TYRO3 was a target of miR­7. The effect of miR­7 and TYRO3 on cell proliferation, migration and invasion was detected in vitro with MTT, wound healing and cell invasion assays, respectively. The expression of proteins associated with the phosphoinositide 3­kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway was detected by western blot analysis. The results indicated that the expression of miR­7 was downregulated and the levels of TYRO3 were gradually increased in CRC tissues and cell lines. TYRO3 was identified as a functional target of miR­7, and its expression was negatively regulated by miR­7 in CRC cell lines. The functional studies demonstrated that miR­7 knockdown promoted the proliferation, migration and invasion of CRC cells, while knockdown of TYRO3 repressed these biological processes. In addition, pathway analyses revealed that the oncogenic effect of TYRO3 was associated with PI3K/AKT/mTOR pathway inhibition. In conclusion, the data suggested that miR­7 promoted the development of CRC by targeting oncogenic TYRO3, which may be mediated by inhibition of the PI3K/AKT/mTOR signaling pathway. Thus, miR­7 may serve as an independent prognostic biomarker in patients with CRC.

Overexpression of RAB34 correlates with poor prognosis and tumor progression in hepatocellular carcinoma.

RAB34, a protein belonging to the RAB family, is involved in protein transport, repositioning of lysosomes and activation of micropinocytosis. However, few studies have reported its function in human epithelial cancers. Immunohistochemistry (IHC) and western blotting were used to detect expression of RAB34 at the tissue and cell levels. Cell Counting Kit-8 (CCK-8), EDU assay and flow cytometry were used for analyzing cell proliferation. Transwell and scratch wound healing assays were used for assessing cell migration ability. Western blotting was used for detecting expression of E-cadherin and N-cadherin. In the present study, we found that both DNA copy and protein level of RAB34 were upregulating in human hepatocellular carcinoma (HCC) tissues when compared with that in adjacent tissues. Analysis of the correlation between RAB34 expression and clinicopathological features showed that patients with overexpression of RAB34 consistently had large tumor size, vessel invasion and poor tumor grade. Furthermore, overall survival analysis showed that patients with upregulated expression of RAB34 were associated with poor prognosis. Moreover, cell function experiments showed that suppression of RAB34 led to a lower proliferation rate and migration ability. In addition, this phenomenon may be attributed to cell cycle phase G1 arrest and mesenchymal-epithelial transition under condition of RAB34 suppression. The present study demonstrated that RAB34 plays an important role in the initiation and progression of HCC. Our results suggest a new therapeutic target for the clinical treatment of HCC.

TSG101 Silencing Suppresses Hepatocellular Carcinoma Cell Growth by Inducing Cell Cycle Arrest and Autophagic Cell Death.

BACKGROUND: The tumor susceptibility gene 101 (TSG101) was originally identified as a tumor-suppressor gene that mediates many molecular and biological processes, such as ubiquitination, endosomal trafficking, cell survival, and virus budding, but its role in hepatocellular carcinoma (HCC) is currently unknown. MATERIAL AND METHODS: We assessed the expression of TSG101 in HCC and paracancerous tissues using qPCR. Then, we used the TSG101-specific siRNA mix to disrupt the expression of TSG101 to investigate the subsequent effect on human hepatoma-7 (Huh7) cells. Western blot was used to detect the protein expression of TSG101 and other molecules. Cell growth assay was performed using CCK8. Transwell assay was used to investigate the migration and invasion ability of Huh7 cells after transfection with of TSG101 siRNA. Flow cytometry was used to estimate the effect of TSG101 knockdown on cell cycle and apoptosis. Confocal laser scanning microscopy was used to observe the actin filaments change and the formation of autophagy. RESULTS: TSG101 was over-expressed in HCC tissues. TSG101 silence was able to suppress Huh7 cell proliferation, migration, and invasion. Furthermore, silencing of TSG101 could induce cell cycle arrest at G1 phase and inhibit the expression of cyclin A and cyclin D, while up-regulating the expression of CDK2. The mechanism might be induction of autophagic cell death and inactivation of Akt and ERK1/2. CONCLUSIONS: TSG101 plays an important role in the development of HCC and may be a target for molecular therapy.

Inhibition of STAT3/cyclinD1 pathway promotes chemotherapeutic sensitivity of colorectal caner.

BACKGROUND: Chemotherapeutic resistance indicated the poor prognosis of colorectal cancer. OBJECTIVE: Our study aimed to investigate the role of STAT3/cyclinD1 pathway in the chemotherapeutic resistance of colorectal cancer. METHODS: We firstly measured the expression of cyclinD1 in the colorectal cancer tissues using immunohistochemistry in tissue microarray. Then cell viability and apoptosis were investigated in the HT-29 cell lines dealing with recombinant lentivirus and shRNA to increase or decrease cyclinD1 expression. Furthermore, luciferase and ChIP assays were applied to investigate whether STAT3 regulated cyclinD1 expression by binding to its promoter. Finally, we determined whether inhibition of STAT3 could decrease cyclinD1 and increase the chemotherapy sensitivity. RESULTS: CyclinD1 expression was significantly increased in the cancer cells and high level of cyclinD1 indicated the poor prognosis. Inhibition of cyclinD1 decreased the cell viability assessed by MTT and increased rate of apoptosis when exposed to 5-FU treatment while overexpression of cyclinD1 showed the reverse effect. ChIP assay showed that STAT3 directly bind to cyclinD1 promoter. Subclone of full promoter of cyclinD1 into pGL4 increased the luciferase activity while delete or mutation of any of STAT3 binding sites resulted in reductions of luciferase activity. Inhibition of STAT3 decreased cyclinD1 expression to decrease the cell viability and increase rate of apoptosis when exposed to 5-FU treatment. CONCLUSIONS: Inhibition of STAT3/cyclinD1 pathway increased the sensitivity of colorectal cancer cell to chemotherapy.