miR-10b-5p Suppresses the Proliferation and Invasion of Primary Hepatic Carcinoma Cells by Downregulating EphA2

Objective. To analyze the function of miR-10b-5p in suppressing the invasion and proliferation of primary hepatic carcinoma cells by downregulating erythropoietin-producing hepatocellular receptor A2 (EphA2). Material and Methods. Eighty-six hepatic carcinoma (HCC) tissue specimens and 86 corresponding adjacent tissue specimens were collected, and the mRNA expression of miR-10b-5p and Ephrin type-A receptor 2 (EphA2) in the specimens was determined using a reverse transcription-polymerase chain reaction (RT-PCR) assay. Western blot was employed to quantify EphA2, B-cell chronic lymphocytic leukemia/lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-3 in the cells, and CCK8, Transwell assay, and flow cytometry were applied to evaluate the proliferation, invasion, and apoptosis of cells, respectively. Moreover, the dual luciferase reporter assay was utilized for correlation analysis between miR-10b-5p and EphA2. Results. miR-10b-5p was lowly expressed in HCC, while EphA2 was highly expressed. Cell experiments revealed that miR-10b-5p overexpression or EphA2 knockdown could reduce cell proliferation, accelerate apoptosis, strongly upregulate Bax and Caspase-3, and downregulate Bcl-2. In contrast, miR-10b-5p knockdown or EphA2 overexpression gave rise to reverse biological phenotypes. Furthermore, dual luciferase reporter assay verified that miR-10b-5p was a target of EphA2, and the rescue experiment implied that transfection of pCMV-EphA2 or Si-EphA2 could reverse EphA2 expression and cell biological functions caused by miR-10b-5p overexpression or knockdown. Conclusions. miR-10b-5p reduced HCC cell proliferation but accelerate apoptosis by regulating EphA2, suggesting it has the potential to be a clinical target for HCC.

The Association between Long Noncoding RNA over Expression and Poor Prognosis of Liver Cancer: A Meta-Analysis

Background. Long noncoding RNA (lncRNA) is considered to be a mediator of carcinogenesis, which may be associated with liver cancer survival. However, the relationship remains inconclusive. Meta-analysis was conducted to analytically review the association between the lncRNA expression level and clinicopathological characteristics and prognostic value of hepatic carcinoma. Materials and Methods. Four databases including Embase, PubMed, Web of Science, and the Cochrane Library were searched to collect studies about the relation between lncRNA overexpression and prognosis of liver cancer, dating from the earliest records of these databases to March 2021. Two researchers independently screened the data and literature to perform a stringent evaluation of the quality of material involved in the study. Meta-analysis was performed by Stata 16.0 software on 42 case-control studies with 6293 samples. Results. The outcomes of meta-analysis are presented as follows: lncRNA overexpression patients had later TNM stage (OR = 0.36, 95% CI (0.31, 0.41), P < 0.001), lower histological grade (OR = 0.56, 95%CI (0.49, 0.65), P < 0.001), more vascular invasion (OR = 2.02, 95% CI (1.74, 2.35), P < 0.001), bigger tumor size (OR = 2.28, 95% CI (2.00, 2.60), P < 0.001), more severe liver cirrhosis (OR = 1.39, 95% CI(0.1.16, 1.66), P < 0.001), more likely to metastasize (OR = 1.80, 95%CI(1.49, 2.18), P < 0.001), and more tumor numbers (OR = 0.72, 95% CI (0.62, 0.84), P < 0.05). lncRNA over expression patients had shorter OS (HR = 2.32, 95 CI% (2.08, 2.59), P < 0.01, RFS (HR = 2.19, 95 CI% (1.72, 2.78), P < 0.01), and DFS (HR = 2.01, 95 CI% (1.57, 2.57), P < 0.01). Conclusions. Overexposure of lncRNA is a poor prognostic feature for patients with hepatic carcinoma. The scope of our study was limited because of a lack of relevant research and the poor representativeness and varying quality of the studies involved in the current meta-analysis. Our conclusion still requires higher studies for further validation. This trial is clinically registered with CRD4201920620.

The relationship between serum hepatitis B core-related antigen and hepatitis B virus DNA in treatment-naïve patients with hepatitis B cirrhosis: A cross-sectional study

Aims. The relationship between hepatitis B core-related antigen (HBcrAg) and hepatitis B virus (HBV) DNA has already been adequately researched in patients with chronic hepatitis B (CHB) infection, but there are only a few researches yet on the correlations between HBcrAg and HBV DNA in treatment-naïve patients with hepatitis B cirrhosis. Here we explore the correlation between HBcrAg and HBV DNA in this population. Methods. Available data and samples of 98 untreated patients with hepatitis B cirrhosis between October 2018 and October 2019 were analysed. Statistical analyses included baseline characteristics, univariate analysis, stratification analysis, three different analytical models, and a generalized additive model. Results. After adjusting for all recorded confounders (sex, age, diagnosis of primary hepatic carcinoma, total bilirubin（TBIL）, hepatitis B surface antigen (HBsAg), hepatitis B e antigen(HBeAg), Child–Pugh class, family history of HBV infection, family history of hepatocellular carcinoma(HCC), alcohol-related liver disease (ALD), and diabetes mellitus), a linear relationship was detected between HBcrAg and HBV DNA (β=0.59, 95%CI=0.34–0.84, P<0.0001). The variational trend of HBcrAg and HBV DNA in each stratified variable (sex, age, HBeAg, family history of HBV infection, family history of HCC, diabetes mellitus, diagnosis of primary hepatic carcinoma, Child–Pugh class, and ALD) were consistent. Conclusion. There was a linear and positive correlation between HBcrAg and HBV DNA in treatment-naïve patients with hepatitis B cirrhosis.

miR-4454 Promotes Hepatic Carcinoma Progression by Targeting Vps4A and Rab27A

Hepatocellular carcinoma (HCC) has high morbidity and mortality. MicroRNAs (miRNAs), which could be regulated by cancer-derived exosomes, play critical regulatory roles in the initiation and development of cancer. However, the expressions, effects, and mechanisms of abundant miRNAs regulated by HCC cancer-derived exosomes in HCC remain largely unclear. Exosomes of HepG2 cells under heat shock, TGF-β1, doxorubicin, acid and hypoxia/reoxygenation (H/R) conditions, and exosomes were successfully identified by transmission electron microscopy and Western blot analysis. The identified exosomes were then applied to evaluate the miRNA expression profiles by RNA sequencing. Mechanically, we discovered that doxorubicin was upregulated, TGF-β1 downregulated the expressions of Vps4A, Rab27A, Alix, and Hrs in HepG2 cells and exosomes, and Vps4A and Rab27A, as target genes for miR-4454, could also be downregulated by miR-4454. Functionally, we revealed that miR-4454 inhibitor and miR-4454 inhibitor-mediated exosomes could markedly suppress proliferation, migration, invasion, and vascularization and accelerate cycle arrest, apoptosis, and ROS of HepG2 cells. This study provided many potential HCC cancer-derived exosome-mediated miRNAs in HCC under 5 different stimulus conditions. Meanwhile, we certified that miR-4454 in exosomes could provide a novel and effective mechanism for HCC function.

Counteracting Action of Curcumin on High Glucose-Induced Chemoresistance in Hepatic Carcinoma Cells

Along with direct anticancer activity, curcumin hinders the onset of chemoresistance. Among many, high glucose condition is a key driving factor for chemoresistance. However, the ability of curcumin remains unexplored against high glucose-induced chemoresistance. Moreover, chemoresistance is major hindrance in effective clinical management of liver cancer. Using hepatic carcinoma HepG2 cells, the present investigation demonstrates that high glucose induces chemoresistance, which is averted by the simultaneous presence of curcumin. Curcumin obviated the hyperglycemia-induced modulations like elevated glucose consumption, lactate production, and extracellular acidification, and diminished nitric oxide and reactive oxygen species (ROS) production. Modulated molecular regulators are suggested to play a crucial role as curcumin pretreatment also prevented the onset of chemoresistance by high glucose. High glucose instigated suppression in the intracellular accumulation of anticancer drug doxorubicin and drug-induced chromatin compactness along with declined expression of drug efflux pump MDR-1 and transcription factors and signal transducers governing the survival, aggressiveness, and apoptotic cell death (p53, HIF-1α, mTOR, MYC, STAT3). Curcumin alleviated the suppression of drug retention and nuclear condensation along with hindering the high glucose-induced alterations in transcription factors and signal transducers. High glucose-driven resistance in cancer cells was associated with elevated expression of metabolic enzymes HKII, PFK1, GAPDH, PKM2, LDH-A, IDH3A, and FASN. Metabolite transporters and receptors (GLUT-1, MCT-1, MCT-4, and HCAR-1) were also found upregulated in high glucose exposed HepG2 cells. Curcumin inhibited the elevated expression of these enzymes, transporters, and receptors in cancer cells. Curcumin also uplifted the SDH expression, which was inhibited in high glucose condition. Taken together, the findings of the present investigation first time demonstrate the ability of curcumin against high glucose-induced chemoresistance, along with its molecular mechanism. This will have implication in therapeutic management of malignancies in diabetic conditions.

Sensitive and specific detection of micrometastasis of hepatic carcinoma in peripheral blood by immunomagnetic nanobeads polymerase chain reaction

This study aims to determine the value of miR-133a-3p in diagnosing micrometastasis of hepatic carcinoma (HCC) from peripheral blood (PB) samples and dissecting the molecular mechanism (MM) of micrometastasis. We selected 70 patients with micrometastasis of HCC from our hospital between August 2014 to August 2020 for the research (Res) group and enrolled 55 patients with HCC but without micrometastasis in the control (Con) group. We collected PB samples from each participant and quantified miR-133a-3p in each sample using immunomagnetic nanobeads (IMNBs)-based PCR and RT-PCR. In contrast to the Con group, the Res group presented a significant down-regulation of miR-133a-3p in PB. The IMNBs and RT-PCR showed that miR-133a-3p exhibited the area under the curves (AUCs) of 0.882 and 0.845, specificities of 81.82% and 74.55%, and sensitivities of 85.71% and 88.57%, respectively. Due to its higher specificity and sensitivity, IMNBs were utilized. IMNBs were then used to quantify miR-133a-3p in HCC cells. Additionally, we explored the association of miR-133a-3p with PAF; miR-133a-3p was inversely correlated with AFP (r =−0.614; P <0.001). miR-133a-3p’s targeting association with MMP15 and the corresponding MM was verified and analyzed. The up-regulation of miR-133a-3p levels inhibited the migration and invasion of HCC cells, while its down-regulation yielded the opposite effect. Moreover, miR-133a-3p negatively regulated the protein level of MMP15, the migration and invasion capacity of HCC cells was greatly impacted. In short, miR-133a-3p decreased in HCC micrometastasis. The detection of miR-133a-3p by IMNBs is both specific and sensitive in diagnosing the micrometastasis. miR-133a-3p is of enormous value in diagnosing HCC micrometastasis from PB samples, and its up-regulation can inhibit MMP15 and hinder HCC metastasis.