Identification of a Novel Variant of PDGFC Associated with Nonsyndromic Cleft Lip and Palate in a Chinese Family.

Nonsyndromic cleft lip with or without cleft palate (NSCL/P) accounts for 70% of the total number of patients with cleft lip with or without cleft palate (CL/P) and is the most common type of congenital deformity of the craniomaxillofacial region. In this study, whole exome sequencing (WES) and Sanger sequencing were performed on affected members of a Han Chinese family, and a missense variant in the platelet-derived growth factor C (PDGFC) gene (NM_016205: c.G93T: p.Q31H) was identified to be associated with NSCL/P. Bioinformatic studies demonstrated that the amino acid corresponding to this variation is highly conserved in many mammals and leads to a glutamine-to-histidine substitution in an evolutionarily conserved DNA-binding domain. It was found that the expression of PDGFC was significantly decreased in the dental pulp stem cells (DPSCs) of NSCL/P cases, compared to the controls, and that the variant (NM_016205: c.G93T) reduced the expression of PDGFC. In addition, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that Pdgfc deficiency disrupted NSCL/P-related signaling pathways such as the MAPK signaling pathway and cell adhesion molecules. In conclusion, our study identified a missense variant (NM_016205: c.G93T) in exon 1 of PDGFC potentially associated with susceptibility to NSCL/P.

[Evaluation of the effect of invisible correction and SGTB in the treatment of mandibular retrusion].

PURPOSE: To compare the clinical effects of invisible correction and SGTB in two-stage treatment of mandibular retrusion. METHODS: Eighty-five patients with bony mandibular regression who did not pass the peak of growth and development were selected. Among them, 40 cases were guided by invisible correction and 45 cases were guided by SGTB functional correction. Lateral head X-rays before and after treatment were measured, and the effect before and after treatment was compared by SPSS 16.0 software package. RESULTS: After treatment, the SNB angle and NP-FH (face angle) increased and the ANB angle decreased in both groups, all of which had significant changes(P＜0.001). At T1, the changes of SNB angle, ANB angle and NP-FH angle in the invisible group were smaller than those in the SGTB group(P＜0.05). The difference between MP-FH angle and MP-SN angle before and after treatment was greater in the SGTB group than in the invisible group(P＜0.001). CONCLUSIONS: In patients with receding mandible before the peak of growth and development, the mandible was significantly moved forward after orthodontics. Both invisible orthodontics and SGTB promote changes in the position of the mandible, improves coordination between the upper and lower jaws. In the control of the posterior teeth in the vertical direction, the invisible group is stronger than the SGTB group.

Three-year observations on the effect of different cusp inclinations on the restoration of short maxillary first molar implants: A randomized controlled trial.

Objective: To investigate the effect of different cusp inclination on short implant prosthesis of maxillary first molar after 3 years of weight-bearing in biology and mechanics. Methods: The clinical patients were randomly selected from the database and divided into four groups A, B, C, and D according to the cusp inclination of the maxillary first molar short implant restoration (4.8 mm × 8 mm, Dentium). 20 cases in each group. The cusp inclination was 10 degrees-15 degrees, 15 degrees-20 degrees, 20 degrees-25 degrees, 25 degrees-30 degrees. After 3 years of weight-bearing, cone beam computed tomography (CBCT) and Florida probe were used to measure and observe the height of alveolar bone (H), periodontal probing depth (PD) and modified sulcus bleeding index (MBI). Visual analogue scale (VAS) was used to evaluate the overall satisfaction of patients, and the mechanical complications of each group within 3 years of implant weight-bearing were counted. Results: The H and PD of group D were 1.09 ± 0.23 and 2.19 ± 0.11 respectively, which were significantly higher than those of group A, B and C (p < 0.05). There was no significant difference in MBI between groups A-D (p > 0.05). The VAS scores of group B and group C were 88.36 ± 5.12 and 88.70 ± 4.52 respectively, which were higher than those of group A and group D (p < 0.05). The incidence of food impaction, porcelain collapse and abutment loosening in group D were 40.0%, 25.0% and 15.0% respectively, which were higher than those in group B and C (p < 0.05). Conclusion: The risk of biological and mechanical complications increases after long-term weight-bearing of maxillary first molar short implant prostheses with high cusp inclination. The cusp inclination of short implant prostheses should be designed as low as 25 degrees.

[Changes of the upper airway in children with Class â ¡ mandibular retrusion and snoring during night before and after functional treatment by sagittal-guidance Twin-block appliance].

PURPOSE: To study the changes of dimension and morphology of upper airway in children with ClassⅡ mandibular retrusion after functional treatment by sagittal-guidance Twin-block appliance. METHODS: Cone-beam CT (CBCT) data of upper airway of the subjects were measured by Dolphin 11.5 software and Mimics 17.0 software , and the changes of dimension and morphology of upper airway before and after functional treatment with sagittal-guidance Twin-block(SGTB) appliance were compared. SPSS 16.0 software package was used for data processing. RESULTS: After functional treatment，the volume of total upper airway，nasopharynx airway, oropharynx airway, the sectional area of tip of the epiglotti(TE), the lateral diameter of TE, the base of the epiglottis(EB) significantly increased (P<0.05) in children with SGTB appliance. CONCLUSIONS: SGTB functional treatment is effective in the treatment of skeletal ClassⅡ mandibular retrusion of children by increase of the upper airway and improvement of respiration．.

A three-phase trans-ethnic study reveals B7-H3 expression is a significant and independent biomarker associated with colon cancer overall survival.

BACKGROUND: There have been inconsistent results and conflicting conclusions among the existing prognostic studies of B7-H3 expression in colon cancer patients. Therefore, the association between B7-H3 expression and colon cancer survival has remained largely unclear. METHODS: We performed a three-phase and trans-ethnic prognostic study of B7-H3 expression in colon cancer patients involving perhaps the largest population to date. In the discovery phase, we utilized a Cox proportional hazards model adjusted for covariates to test the association between B7-H3 expression and colon cancer overall survival (OS) time in a European population from The Cancer Genome Atlas (TCGA) cohort (n=433). In the validation phase I, the association was replicated in a European population from Gene Expression Omnibus (GEO) cohort (n=811). In the validation phase II, we again confirmed the significant association in an Asian population from Fujian Medical University Union Hospital (UNION) cohort (n=179). Furthermore, a series of Kaplan-Meier analysis, bioinformatics analysis of tumor immune microenvironment (TIME), and immune checkpoint prognostic prediction analysis, as well as sensitivity analysis, were also conducted. RESULTS: Highly expressed B7-H3 was a significant and robust biomarker to colon cancer survival, with a large hazard ratio (HR) [HRTCGA =4.60, 95% confidence interval (CI): 2.15 to 9.83, P=8.37×10-05; HRGEO =1.47, 95% CI: 1.12 to 1.94, P=0.0056; HRUNION =1.63, 95% CI: 1.36 to 1.95, P=7.91×10-08]. We detected an involvement of B7-H3 in the tumor immune microenvironment (TIME). Meanwhile, B7-H3 was significantly and weakly correlated with 6 out of 27 well-recognized immune checkpoint genes. Even after adjusting for effects of other immune checkpoint genes, B7-H3 still exhibited a harmful effect on colon cancer survival using samples from TCGA and GEO cohorts (HR =1.47, 95% CI: 1.07 to 2.02, P=0.0184), indicating that it was an independent prognostic factor of colon cancer. We also proposed an immune checkpoint prognostic risk score which possessed the capability to identify colon cancers with high risk of mortality. CONCLUSIONS: The expression of B7-H3 is a significant, robust, and independent prognostic factor to colon cancer OS.

Nuclear receptor FXR impairs SK-Hep-1 cell migration and invasion by inhibiting the Wnt/ß-catenin signaling pathway.

Recently, the nuclear receptor farnesoid X receptor (FXR) has been considered to be a liver tumor suppressor. However, the role of FXR in liver cancer invasion and metastasis remains unclear. The results of the current study demonstrated that FXR suppressed the migratory and invasive capacities of SK-Hep-1 cells in vitro and that FXR overexpression inhibited local invasion and lung metastasis of SK-Hep-1 ×enografts in vivo. Bioinformatics analysis of the gene expression profile of SK-Hep-1 cells with different FXR levels indicated that FXR may regulate the Wnt/ß-catenin pathway. Compared with controls, FXR-overexpressing SK-Hep-1 cells exhibited decreased expression of ß-catenin target genes and reduced nuclear translocation of ß-catenin proteins in vitro and in vivo. In conclusion, these results indicated that FXR may suppress SK-Hep-1 cell invasion and metastasis by suppressing the Wnt/ß-catenin signaling pathway. The current study provided novel insight into the diagnosis and treatment of liver cancer.

Activation of NLRP3 inflammasome in hepatocytes after exposure to cobalt nanoparticles: The role of oxidative stress.

With the increased use of nanomaterials and increased exposure of humans to various nanomaterials, the potential health effects of nanomaterials cannot be ignored. The hepatotoxicity of cobalt nanoparticles (Nano-Co) is largely unknown and the underlying mechanisms remain obscure. The purpose of this study was to exam the hepatotoxicity induced by Nano-Co and its potential mechanisms. Our results showed that exposure of human fetal hepatocytes L02 to Nano-Co caused a dose- and a time-dependent cytotoxicity. Besides the generation of reactive oxygen species (ROS) and mitochondrial reactive oxygen species (mtROS), exposure to Nano-Co also caused activation of NOD-like receptor protein 3 (NLRP3) inflammasome in hepatocytes. After silencing NLRP3, one component of NLRP3 inflammasome, expression by siRNA strategy, we found that upregulation of NLRP3-related proteins was abolished in hepatocytes exposed to Nano-Co. Using antioxidants to scavenge ROS and mtROS, we demonstrated that Nano-Co-induced mtROS generation was related to Nano-Co-induced NLRP3 inflammasome activation. Our findings demonstrated that Nano-Co exposure may promote intracellular oxidative stress damage, and mtROS may mediate the activation of NLRP3 inflammasome in hepatocytes exposed to Nano-Co, suggesting an important role of ROS/NLRP3 pathway in Nano-Co-induced hepatotoxicity. These results provide scientific insights into the hepatotoxicity of Nano-Co and a basis for the prevention and treatment of Nano-Co-induced cytotoxicity.

Determination of Phosphorylated Histone H2AX in Nanoparticle-Induced Genotoxic Studies.

DNA double-strand breaks (DSBs), one of the most severe lesions of DNA damage triggered by various genotoxic insults, can lead to chromosome change, genomic instability, and even tumorigenesis if not repaired efficiently. In response to DNA damage, histone H2AX molecules are rapidly phosphorylated at serine 139 near the site of DNA DSBs and form γ-H2AX foci. As an early important cellular event linked to DNA damage and repair, γ-H2AX is a highly sensitive biomarker for "monitoring" DNA damage and consequently is a useful tool in genetic toxicology screen. We and other researchers have used γ-H2AX as a marker to assess the potential genotoxic effects of some nanoparticles in vitro and in vivo. In this chapter, we describe several useful methods for γ-H2AX detection, which can be used to evaluate the potential genotoxic effects of nanoparticles.

[Analysis of the biocompatibility of a new PLGA/fish skin collagen conjugated electrostatic spinning film].

PURPOSE: To study the biocompatibility of a new polylactic acid-glycolic acid(PLGA)/fish skin collagen conjugated electrostatic spinning film. METHODS: PLGA and medical-grade fish skin-I collagen were used as raw materials to prepare nanometer fiber films by conjugated electrostatic spinning technique. Mouse fibroblast L929 was used as a model to evaluate its cell and histocompatibility, which provided experimental basis for application in guided bone regeneration (GBR). RESULTS: Fibroblasts showed good adhesion and growth on the surface of fiber membrane; there were a large number of deep pseudopodias, which were arranged along the orientation of fibers. CONCLUSIONS: PLGA/fish skin collagen conjugated electrostatic spinning membrane is beneficial to fibroblast growth and is expected to be used for GBR.

[Morphological characteristics of upper alveolar bone around central incisor in Class II division 1 and division 2 malocclusion on cone-beam CT].

PURPOSE: During orthodontic treatment, the incisors in patients with Class II division 1 and division 2 malocclusion showed different clinical features, which determine the different ways to move the teeth to the suitable position. This study analyzed the morphology of alveolar bone around upper central incisor with cone-beam CT (CBCT). METHODS: The sample consisted of 20 normal malocclusion and 40 patients with Class II malocclusion （division 1 20 cases and division 2 20 cases）. CBCT images before orthodontic treatment were reconstructed by INVIVO 5.0 image processing software. Paired t test was used for statistical analysis with SPSS 19.0 software package. RESULTS: At the upper central incisors, the alveolar thickness between normal malocclusion and Class II division 1 and division 2 malocclusion had significant difference except the palate thickness between normal malocclusion and Class II division 2 malocclusion (P<0.05). CONCLUSIONS: Orthodontist should consider the relationship between the teeth position and alveolar bone shape comprehensively, choose the correct plan in order to avoid root desorption, alveolar bone loss, fenestration and other adverse reactions.

[Comparison of condylar position between Angle Class I and Class II malocclusion in teenagers].

PURPOSE: To analyze and compare the difference of condylar position between Angle Class I and Class II malocclusion patients using cone-beam CT (CBCT). METHODS: Thirty Class I patients, 30 Class II division 1 patients and 30 Class II division 2 patients were selected in this study. Each patient underwent CBCT. The images in the oblique position perpendicular to the condyloid process were reconstructed by Examvision software. The joint space was measured by Exam Vision software. The data were processed with SPSS17.0 software package. RESULTS: The upper joint space was larger in Class II, the posterior joint space was smaller in Class II patients; and in Class II division 1 patients, both of the upper and anterior joint spaces were larger than in Class II division 2 patients，the differences were significant (P<0.05). The length of condyle was longer in Class I patients than in Class II patients. CONCLUSIONS: The condylar position in Class II division 2 patients was lower and further backward. The length of condyle is shortest in Class II division 2 patients.

[The effect of core veneer thickness ratio on the flexural strength of diatomite-based dental ceramic].

PURPOSE: To evaluate the effect of different core veneer thickness ratios on the flexural strength and failure mode of bilayered diatomite-based dental ceramics. METHODS: Diatomite-based dental ceramics blocks (16 mm×5.4 mm×1 mm) were sintered with different thickness of veneer porcelains: 0 mm (group A), 0.6 mm (group B), 0.8 mm (group C) and 1.0 mm (group D). Flexural strength was detected and scanning electron microscope was used to observe the interface microstructure. Statistical analysis was performed using SPSS 17.0 software package. RESULTS: With the increase of the thickness of the veneer porcelain, flexural strength of group C showed highest flexural strength up to (277.24±5.47) MPa. CONCLUSIONS: Different core veneer thickness ratios can significantly influence the flexural strength of bilayered diatomite-based dental ceramics. Supported by Science and Technology Projects of Nantong City (HS2013010).

[A study on the thickness of buccal bone in the mandible of different vertical facial type in adults using cone-beam CT].

PURPOSE: To provide some references of using micro-implant anchorage in clinical orthodontic treatment, the thickness of buccal bone of mandible of different vertical facial type in adults with cone-beam CT (CBCT) were measured. METHODS: Initial 3-dimensional images of 45 adult patients (15 men, 30 women)were reoriented by using a standardized protocol, and divided into 3 groups by angle of mandibular plane (G1: high angle, 15 patients; G2: average angle, 15 patients; G3: low angle, 15 patients). After signing the informed consent form, three measurement points were defined at 4, 6, and 8mm from the alveolar crest in each measurement area. Statistical analysis was performed with SPSS11.0 software package. RESULTS: The thickness of buccal bone in mandible was thinner in G1 than in G2, and was thinnest in G3. The cortical bone thickness was thickest at the 8 mm level and thinnest at the 4mm level. CONCLUSIONS: The study suggested that the cortical bone thickness at 4mm level from the alveolar crest was safe for implantation. It is more stable when the distance of implants was more away from the alveolar crest. Patients with low mandibular plane angle should use self-tapping micro-implant anchorage. Supported by Nantong Science and Technology Fund (HS2013033).

[Preliminary study of bonding strength between diatomite-based dental ceramic and veneering porcelains].

PURPOSE: In order to choose the best veneering porcelain for diatomite-based dental ceramic substrate, the bonding strength between diatomite-based dental ceramics and veneering porcelains was measured, and the microstructure and elements distribution of interface were analyzed. METHODS: The coefficient of thermal expansion (CTE) of diatomite-based dental ceramics was detected by dilatometry. Three veneering porcelain materials were selected with the best CTE matching including alumina veneering porcelain (group A), titanium porcelain veneering porcelain (group B), and E-max veneering porcelain (group C). Shear bonding strength was detected. SEM and EDS were used to observe the interface microstructure and element distribution. Statistical analysis was performed using SPSS 17.0 software package. RESULTS: The CTE of diatomite-based dental ceramics at 25-500 degrees centigrade was 8.85×10-6K-1. The diatomite-based substrate ceramics combined best with group C. Shear bonding strength between group A and C and group B and C both showed significant differences(P<0.05). SEM and EDS showed that the interface of group C sintered tightly and elements permeated on both sides of the interface. CONCLUSIONS: The diatomite-based substrate ceramics combines better with E-max porcelain veneer.

Lentiviral-mediated short hairpin RNA silencing of APE1 suppresses hepatocellular carcinoma proliferation and migration: A potential therapeutic target for hepatoma treatment.

Apurinic/apyrimidinic endonuclease-1 (APE1) is a protein involved in DNA repair and transcriptional regulation of gene expression. APE1 expression was reported to be correlated with poor prognosis in hepatocellular carcinoma (HCC) patients. Based on our previous study, we hypothesized that APE1 may be involved in the metastatic progression of HCC. Thus, the present study aimed to investigate the knockdown effect of APE1 using shRNA in HCC and demonstrate that silencing of APE1 in MHCC97-H cells can decrease the oncogenic transforming potential in vitro and reduce the growth of HCC tumor xenografts in vivo. Silencing of APE1 expression decreased the cell proliferation and survival, reduced the cell adhesion ability in Matrigel or fibronectin-coated plates and suppressed the cell migration and invasion in a Transwell assay of HCC cells. In the xenograft study, tumor growth was markedly inhibited in the APE1-silenced group. Silencing of APE1 in MHCC97-H cells decreased the oncogenic transforming potential in vitro and reduced the growth of HCC tumor xenografts in vivo. Inhibition of APE1 may present a novel therapeutic approach for the treatment of HCC.

[Clinical effect of premolar extraction with high torque self-locking appliances].

PURPOSE: To analyze the clinical effect of extraction correction of patients with maxillary protrusion by using Damon high torque appliances. METHODS: Forty patients with extraction of four first premolars due to orthodontic treatment were selected. They had protrusion of maxilla and were randomly divided into two groups. Both arches of Damon group were treated with Damon high torque appliances and MBT group were corrected with traditional MBT straight wire appliance; patients in two groups received sliding mechanics for adduction of upper anterior teeth. Cephalometric radiophotographs were analyzed before（T0）and after adduction of upper anterior teeth（T1）. The difference between the two groups was analyzed with SPSS15.0 software package. RESULTS: The upper incisors showed a more retruded position in the two groups at T1,although they showed a similar amount in both groups. The backward movement of upper lip was 2.31mm in Damon group, which was significantly less than that of 2.81mm in MBT group(P<0.05). No significant difference in the first molar mesialization was detected between Damon group(2.33 mm) and MBT group(2.36mm) (P>0.05). For anterior teeth retraction, nasolabial angle (NLA) in MBT and Damon group were 107.44°and 106.33°, increase of NLA in MBT group was 9.17°,which was significantly larger than that of 7.22°in Damon group (P<0.05). U1-MxP in MBT and Damon were 110.61°and 114.11°. U1-MxP decreased 12.06°in MBT group, while significant change was deteced in Damon group of 7.61°(P<0.05). CONCLUSIONS: Application of Damon high torque appliances in extraction treatment of maxillary protrusion patients yielded better control of the labial crown torque of anterior teeth, and avoided excessive lingual inclination of anterior teeth compared with MBT in adduction of upper anterior teeth.

[Morphology and microRNA expression profiles of drug-resistant cells in hepatocellular carcinoma].

OBJECTIVE: To compare morphological differences of three drug-resistant hepatocellular carcinoma (HCC) cell subclones (Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) and their parental Huh-7 cell line, to analyze differential microRNA (miRNA) expression profiles in these cells and, finally to screen for the abnormal expressed miRNAs in drug-resistant HCC cells. METHODS: Cellular morphology was observed by histology and transmission electron microscopy. MiRNA microarray was used to analyze the differential miRNA expression profiles in these cells (Huh-7, Huh-7/ADM, Huh-7/CBP, Huh-7/MMC) followed by real time quantitative PCR validation. RESULTS: The drug-resistant cells had more intracytoplasmic organelles and were larger in size along with increased cytological pleomorphism than the parental Huh-7 cells. Compared with the parental Huh-7 cells, 32 simultaneously up-regulated and 22 down-regulated miRNAs were found in three drug-resistant cells. Up-regulation of miR-15a, miR-16, miR-27b, miR-30b, miR-146a, miR-146b-5p, miR-181a, miR-181d and miR-194 was verified by RT-qPCR. CONCLUSION: Drug-resistant HCC cells have abnormal expressed miRNAs, which may be explored to further investigate the association of miRNA expressions with multidrugs resistance in HCC.

Potential therapeutic value of dendritic cells loaded with NY­ESO­1 protein for the immunotherapy of advanced hepatocellular carcinoma.

NY­ESO­1 is one of the most immunogenic cancer-testis (CT) antigens. Cancer vaccine trials based on NY­ESO­1 are currently ongoing. Dendritic cells (DCs) are the most potent antigen-presenting cells. The immune functions of DCs in a number of tumors have been identified; however, the potential therapeutic value of DCs pulsed with NY­ESO­1 in hepatocellular carcinoma (HCC) has not been extensively investigated. The objectives of the present study were to evaluate T cell response following stimulation with DCs pulsed with the recombinant NY­ESO­1 protein (rESO) and to establish a correlation between NY­ESO­1 expression and clinicopathological features in HCC patients. DCs were generated with granulocyte/macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL­4) from human peripheral blood mononuclear cells. A mixed T cell reaction with DCs loaded with recombinant NY­ESO­1 protein (rESO-DCs) was evaluated by MTT assay. T cell responses against HCC cell lines were analyzed by measuring lactate dehydrogenase (LDH) activity. The protein levels of NY­ESO­1 were detected by immunohistochemistry (IHC) in a tissue microarray (TMA) containing 190 HCC samples. NY­ESO­1 transcript abundance was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) in 54 out of the 190 HCC samples. The results revealed that mature DCs were induced and that rESO­DCs significantly stimulated T cell proliferation. The specific lysis of T cells stimulated with rESO­DCs was significantly higher in the NY­ESO­1-positive HCC cells compared with the NY­ESO­1-negative cells and the other controls (p<0.01). NY­ESO­1 was expressed in 15.8% (30/190)of the HCC samples, as shown by IHC and in 24.1% (13/54) of the samples, as shown by RT-PCR. The frequency of NY­ESO­1 expression was significantly higher in HCC patients with portal vein tumor thrombosis (24.6%) compared with those without thrombosis (11.2%, p=0.013). Our data suggest that DCs loaded with NY­ESO­1 protein stimulate antigen-specific T cell responses against HCC cells in vitro. NY­ESO­1 may thus be used as a potential target for immunotherapy in advanced HCC.

Differential miRNA expression profiles in hepatocellular carcinoma cells and drug-resistant sublines.

Altered expression of miRNAs may contribute to multidrug resistance (MDR) in human cancers. This study investigated the association between miRNAs and MDR in five different drug-resistant hepatocellular carcinoma (HCC) cell sublines. The HCC Huh-7 cell line was treated with adramycin (ADM), cisplatin (DDP), carboplatin (CBP), mitomycin C (MMC) or vincristine (VCR) at increasing concentrations to develop drug-resistant sublines. The cell viability MTT assay was used to detect drug resistance. Five different drug-resistant HCC sublines, Huh-7/ADM, Huh-7/CBP, Huh-7/DDP, Huh-7/MMC and Huh-7/VCR, were established. Cells that were resistant to one drug were also found to be resistant to the other drugs. miRNA microarrays were analyzed to identify differential miRNA expression profiles in these cell lines, and real-time PCR was used to validate miRNA microarray data. miRNA microarray analysis showed that there were 53 upregulated miRNAs in Huh-7/ADM, 56 in Huh-7/CBP, 58 in Huh-7/DDP, 58 in Huh-7/MMC and 49 in Huh-7/VCR, whereas there were 52 downregulated miRNAs in Huh-7/ADM, 50 in Huh-7/CBP, 41 in Huh-7/DDP, 55 in Huh-7/MMC and 56 in Huh-7/VCR. Moreover, 26 simultaneously upregulated and 25 simultaneously downregulated miRNAs were noted in the Huh-7/ADM, Huh-7/CBP, Huh-7/DDP and Huh-7/MMC sublines compared to the parental Huh-7 cell line. In contrast, among these 51 upregulated and downregulated miRNAs, 12 miRNAs were upregulated and 13 miRNAs were downregulated in Huh-7/VCR. Upregulation of miR-27b, miR-181a, miR-146b-5p, miR-181d and miR-146a expression was verified using real-time RT-PCR in the parental and the five drug-resistant cell lines. In conclusion, the present study demonstrates that the differentially expressed miRNA profiles in these five drug-resistant HCC sublines could be useful to further investigate the association of miRNA expression with drug resistance in HCC.