Human Hepatocellular Cancer Research Articles

The Antiproliferative Activity of Oxypeucedanin via Induction of G2/M Phase Cell Cycle Arrest and p53-Dependent MDM2/p21 Expression in Human Hepatoma Cells.

Oxypeucedanin (OPD), a furocoumarin compound from Angelica dahurica (Umbelliferae), exhibits potential antiproliferative activities in human cancer cells. However, the underlying molecular mechanisms of OPD as an anticancer agent in human hepatocellular cancer cells have not been fully elucidated. Therefore, the present study investigated the antiproliferative effect of OPD in SK-Hep-1 human hepatoma cells. OPD effectively inhibited the growth of SK-Hep-1 cells. Flow cytometric analysis revealed that OPD was able to induce G2/M phase cell cycle arrest in cells. The G2/M phase cell cycle arrest by OPD was associated with the downregulation of the checkpoint proteins cyclin B1, cyclin E, cdc2, and cdc25c, and the up-regulation of p-chk1 (Ser345) expression. The growth-inhibitory activity of OPD against hepatoma cells was found to be p53-dependent. The p53-expressing cells (SK-Hep-1 and HepG2) were sensitive, but p53-null cells (Hep3B) were insensitive to the antiproliferative activity of OPD. OPD also activated the expression of p53, and thus leading to the induction of MDM2 and p21, which indicates that the antiproliferative activity of OPD is in part correlated with the modulation of p53 in cancer cells. In addition, the combination of OPD with gemcitabine showed synergistic growth-inhibitory activity in SK-Hep-1 cells. These findings suggest that the anti-proliferative activity of OPD may be highly associated with the induction of G2/M phase cell cycle arrest and upregulation of the p53/MDM2/p21 axis in SK-HEP-1 hepatoma cells.

Simultaneous and ultrasensitive detection of multiple microRNAs by single-molecule fluorescence imaging.

Cell status changes are typically accompanied by the simultaneous changes of multiple microRNA (miRNA) levels. Thus, simultaneous and ultrasensitive detection of multiple miRNA biomarkers shows great promise in early cancer diagnosis. Herein, a facile single-molecule fluorescence imaging assay was proposed for the simultaneous and ultrasensitive detection of multiple miRNAs using only one capture anti-DNA/RNA antibody (S9.6 antibody). Two complementary DNAs (cDNAs) designed to hybridize with miRNA-21 and miRNA-122 were labelled with Cy3 (cDNA1) and Cy5 (cDNA2) dyes at their 5′-ends, respectively. After hybridization, both miRNA-21/cDNA1 and miRNA-122/cDNA2 complexes were captured by S9.6 antibodies pre-modified on a coverslip surface. Subsequently, the Cy3 and Cy5 dyes on the coverslip surface were imaged by the single-molecule fluorescence setup. The amount of miRNA-21 and miRNA-122 was quantified by counting the image spots from the Cy3 and Cy5 dye molecules in the green and red channels, respectively. The proposed assay displayed high specificity and sensitivity for singlet miRNA detection both with a detection limit of 5 fM and for multiple miRNA detection both with a detection limit of 20 fM. Moreover, it was also demonstrated that the assay could be used to detect multiple miRNAs simultaneously in human hepatocellular cancer cells (HepG2 cells). The proposed assay provides a novel biosensing platform for the ultrasensitive and simple detection of multiple miRNA expressions and shows great prospects for early cancer diagnosis.

Metoclopramide increased cell proliferation in HepG2 cell line and sorafenib attenuated the effect

Aim: Metoclopramide is an antiemetic drug used for treating postoperative or chemotherapy-induced emesis. Sorafenib is a kinase inhibitor drug and is approved for the treatment of advanced primary liver cancer, renal cell carcinoma, thyroid cancer, and acute myeloid leukemia. Hepatocellular cancer is a common cause of cancer-related death and its treatment may require coadministration of an antiemetic medication. The study aims to investigate the effect of metoclopramide on hepatocellular cancer cell proliferation alone or in combination with sorafenib. Material and Methods: Metoclopramide doses of 0.17 µM to 25 µM alone or in combination with sorafenib were administered to human hepatocellular cancer cell line, HepG2. Cell viability and proliferation test was used to determine the possible effects on proliferation. Hematoxylin and Eosin staining was performed to visualize the morphological effects of the treatments. Results: Metoclopramide doses of 0.58 µM, 25µM increased cell proliferation when compared to the control group. Metoclopramide combination groups with 9.9 µM sorafenib were compared with control and sorafenib groups. Each combination group was comparable with the control group. Metoclopramide increased proliferation in certain doses. Safety concerns about its use in hepatocellular cancer should be confirmed in clinical trials Each combination group was comparable with the control group. Conclusion: Metoclopramide increased proliferation in certain doses. Sorafenib inhibited the effect. Safety concerns about its use in hepatocellular cancer should be addressed in clinical trials.

The effect of inhibitors of phosphatidylinositol 3-kinase-related kinases on dibenzo[def,p]chrysene genotoxicity measured by γH2AX levels and neutral comet assay in HepG2 human hepatocellular cancer cells

In the study the modulating effect of inhibition of phosphatidylinositol 3-kinase-related kinases (PIKK): ATM (Ataxia Telangiectasia Mutated), ATR (Ataxia Telangiectasia and Rad3 Related) and DNA-PK (DNA-dependent protein kinase) on genotoxicity of dibenzo[def,p]chrysene (DBC) in HepG2 human hepatocellular cancer cells was investigated. The cytotoxicity of DBC was determined, also in combination with PIKK inhibitors, using the MTT reduction assay. The high cytotoxicity of DBC was observed after 72 h incubation (IC50 = 0.06 μM). The PIKK inhibitors applied at non-cytotoxic concentrations: caffeine (1 mM) and KU55933 (2.5 μM) had no significant influence on the DBC cytotoxicity, however NU7026 (5 μM) caused significant increase in the cell viability by about 25%. The combinations of the inhibitors (double or triple) where NU7026 was present also caused increase in the cell viability (i.e. cytoprotective effect) compared to the effect of DBC. The level of damage to the genetic material (DNA double strand breaks, DSB) was assessed by measuring levels of phosphorylated form of H2A histone (γH2AX) and neutral comet assay. DBC induced DSB in a concentration and time-dependent manner. NU7026 considerably reduced the level of DSB level measured by γH2AX and comet assay.The obtained results confirm that DBC is cytotoxic and causes damage to the genetic material including DSB. The DNA-PK inhibitor NU7026 increases cell viability after exposure to DBC and reduces DNA damage, what indicates an important role of the sensor kinase in mediating the effect.

Clinical significance and biological function of co-stimulation molecule B7 homolog 6 expression in human hepatocellular carcinoma

Objective To investigate the clinical significance and biological function of co-stimulatory molecule B7 homolog 6 (B7-H6) in human hepatocellular cancer. Methods The immunohistochemistry assay and the tissue micro-array were used to examine the B7-H6 expression in hepatocellular cancer tissues. The correlation between the expression level of B7-H6 and patients’ clinic pathological parameters was evaluated by using chi-square test. Down-regulation of B7-H6 was performed by using RNA interference (RNAi) method to investigate the role of B7-H6 in regulation of biological behaviors in human hepatocellular cancer cell lines HepG2 and SMMC-7721. The cell counting kit-8 (CCK-8) assay, wound healing assay, and Transwell assay were used to examine the cellular function after B7-H6 knockdown of these cells. Results The B7-H6 expression level in hepatocellular cancer tissues was significantly correlated with patients’ age (χ2=5.943, P<0.05) and tumor size (χ2=4.519, P<0.05). Next, to investigate the functions of B7-H6 in human hepatocellular cancer, we successfully constructed B7-H6 knockdown expression human hepatocellular cell lines using the RNA interference technology. CCK-8 assay showed that decreased B7-H6 expression in human hepatocellular cancer lines HepG2 and SMMC-7721 significantly attenuated cell proliferation (HepG2, 48 h: 0.413±0.020 vs. 0.572 ±0.025, t=4.951, P<0.05; 72 h: 0.503±0.014 vs. 0.691±0.018, t=8.372, P<0.01; SMMC-7721, 48 h: 0.534± 0.001 vs. 0.656±0.021, t=5.869, P<0.05; 72 h: 0.650± 0.044 vs. 0.856±0.025, t=4.069, P<0.05). The wound healing assay showed that decreased B7-H6 expression in human hepatocellular cancer lines HepG2 and SMMC-7721 significantly attenuated cell migration (HepG2, 24 h: t=4.581, P<0.05, 36 h: t=5.040, P<0.01; SMMC-7721, 36 h: t=5.118, P<0.01). The Transwell assay showed that decreased B7-H6 expression in human hepatocellular cancer lines HepG2 and SMMC-7721 significantly attenuated cell invasion (HepG2, 24 h: t=6.824, P<0.01, 36 h: t=11.030, P<0.001; SMMC-7721, 24 h: t=10.180, P<0.001, 36 h: t=10.340, P<0.001). Conclusion B7-H6 had important effect in initiation and development of human hepatocellular cancer. Key words: B7 homolog 6; Liver cancer; Immunohistochemistry; RNA interference; Tissue microarray

Protein kinase C inhibitors override ZEB1-induced chemoresistance in HCC

Epithelial–mesenchymal transition (EMT) is a process by which tumour cells lose epithelial characteristics, become mesenchymal and highly motile. EMT pathways also induce stem cell features and resistance to apoptosis. Identifying and targeting this pool of tumour cells is a major challenge. Protein kinase C (PKC) inhibition has been shown to eliminate breast cancer stem cells but has never been assessed in hepatocellular cancer (HCC). We investigated ZEB family of EMT inducer expression as a biomarker for metastatic HCC and evaluated the efficacy of PKC inhibitors for HCC treatment. We showed that ZEB1 positivity predicted patient survival in multiple cohorts and also validated as an independent biomarker of HCC metastasis. ZEB1-expressing HCC cell lines became resistant to conventional chemotherapeutic agents and were enriched in CD44high/CD24low cell population. ZEB1- or TGFβ-induced EMT increased PKCα abundance. Probing public databases ascertained a positive association of ZEB1 and PKCα expression in human HCC tumours. Inhibition of PKCα activity by small molecule inhibitors or by PKCA knockdown reduced viability of mesenchymal HCC cells in vitro and in vivo. Our results suggest that ZEB1 expression predicts survival and metastatic potential of HCC. Chemoresistant/mesenchymal HCC cells become addicted to PKC pathway and display sensitivity to PKC inhibitors such as UCN-01. Stratifying patients according to ZEB1 and combining UCN-01 with conventional chemotherapy may be an advantageous chemotherapeutic strategy.

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis.

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Regulation of apoptosis and autophagy by luteolin in human hepatocellular cancer Hep3B cells

In several cancer cells, luteolin (3′,4′,5,7-tetrahydroxyflavone) exerts anticancer effects by upregulation of oxidative stress and endoplasmic reticulum (ER) stress, which are shown to activate p53-dependent cell death. Since luteolin-mediated ER stress regulation has not been investigated in hepatocellular carcinoma (HCC) cells, we investigated the role of ER stress in anti-carcinogenic effects using p53-wild type and p53-null HCC cells treated with luteolin. Trypan blue exclusion test was implemented to determine cell viability. Western blot was applied to compare the difference of autophagy, apoptosis, and proliferation event between cell lines. ER stress and p53 activation were determined by RT-PCR. Our results showed that luteolin at 5–10 μmol/L induced higher cytotoxicity in p53-null Hep3B cells than in p53-wild type HepG2 cells. Cytotoxicity was not observed in normal liver cells. Apoptosis activation and proliferation inhibition occurred in only p53-null Hep3B cells in response to luteolin treatment. Also, luteolin induced oxidative stress and ER stress in p53-null Hep3B cells. Although we observed the induction of p21 in Hep3B cells, the concomitant increases in mRNA levels of p53 family members, including TAp63 and TAp73, were not observed. Furthermore, luteolin induced autophagy in Hep3B cells only, which enhanced cell viability. Taken together, this study suggests that luteolin-induced ER stress may exert anticancer effects in a p53-independent manner.

Suberin/trans-Cinnamaldehyde Oil Nanoparticles with Antimicrobial Activity and Anticancer Properties When Loaded with Paclitaxel.

Suberin is a biopolyester that can be extracted from bioresources such as cork trees and may find many applications in food packaging, sealing, and biomedical fields. In this work, we describe for the first time the preparation of suberin nanoparticles using the solvent/antisolvent method. Three different suberin extraction methods from cork trees were used to reveal any differences in the production and properties of the nanoparticles. trans-Cinnamaldehyde, a component of cinnamon essential oil, was added to suberin nanoparticles due to its antimicrobial and/or anticancer properties, and its synergistic effect was studied with the inherent biological properties of suberin. Interestingly, both suberin and suberin/trans-cinnamaldehyde nanoparticles showed very good antimicrobial properties against Staphylococcus aureus and Pseudomonas aeruginosa bacteria, and also against Candida albicans (yeast). Suberin nanoparticles containing trans-cinnamaldehyde loaded with 0.1% w/w paclitaxel provided significant anticancer properties against human hepatocellular carcinoma HepG2 cancer cell lines. This study can be used to explore novel biobased polymers as efficient carriers of essential oils and drugs for infection and cancer therapy, by using minimum amount of drugs and therefore reducing their cytotoxicity and side effects.

Abstract 4321: Epigenetic regulation of TP73 expression in HCC and GI cancer

Abstract Tumorigenesis results from an accumulation of mutational and epigenetic changes that alter normal cell growth and survival pathways. TP73 is the homologue of the master tumor suppressor TP53 involved in cellular responses to stress and development. The TP73 gene encodes two different protein isoforms, TAp73 and ΔNp73, and maps to a region on chromosome 1p36 that is frequently deleted in neuroblastoma and other tumors, and is thought to contain multiple tumor suppressor genes. However, the analysis of p73-knockout mice yielded conflicting results with respect to tumor suppression. World-wide efforts in sequencing the TP73 gene in patient tumor samples have not provided evidence for genetic alterations as a common cause of TP73 inactivation in human cancer. The role of TP73 in tumorigenesis has remained elusive to date. In this study, we isolated two stem cell lines from normal young and old human liver tissues to determine TP73 expression in normal human liver stem cell and hepatocellular cancer (HCC) cell lines (HepG2, SNU398, SNU449 &amp; SNU475), gastrointestinal cancer (GI) cell lines (Caco2 &amp; HCT116) and a normal skin fibroblast cell line (HS27). The results show that TP73 only expresses in cancer cell lines. Further immunohistochemistry studies on human normal liver tissues as well as adjacent normal and cancerous liver tissues from HCC patients confirm that TP73 only expresses in the cancerous tissues. Moreover, methylation-specific PCR and bisulfite sequencing studies revealed that TP73 promoter is activated only in cancer cell lines by DNA methylation. Furthermore, ChIP assay results demonstrated that CTCF (a chromosomal networking protein CCCTC binding factor) binds to TP73 promoter and regulates TP73 expression. Our observation may prove significant for the development of future therapeutic and diagnostic applications. Citation Format: Zhixing Yao, Cristina Di Poto, Habtom W. Ressom, Zaki A. Sherif. Epigenetic regulation of TP73 expression in HCC and GI cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4321.

Abstract 4960: Studying the impact of gut microbiome to hepatocellular cancer in an orthotopic murine model

Abstract Background and Objective: As one of the most important components of human body, gut microbiota play a vital role in human health by participating in metabolomics and maintaining immunological homeostasis. Microbiota dysbiosis is significantly associated with initiation and progression of different diseases. Due to close anatomical and functional connection between gut and liver (gut-liver axis), gut microbiota are found to impact the development of liver diseases including hepatocellular cancer (HCC) without fully understanding mechanisms. The overall objective of present study is to investigate how microbiota modulate HCC growth by regulating intrahepatic immune response. Methods: Administration of carbon tetrachloride in combination with transplant of oncogenic hepatocytes by intrasplenic injection was used to make an orthotopic mouse model of HCC. A specific antibiotics cocktail (ABX) was selected to inhibit some kinds of gut microbiota. 16s rRNA sequencing was used to characterize gut microbiota profile in mice with the different treatments. Magnetic resonance imaging (MRI) was used to monitor tumor growth. Adoptive transfer of tumor antigen-specific CD8+ T cells following the treatment was used to investigate how ABX administration impacted intrahepatic immune response. Flow cytometry and multiplex cytokine assay were used to detect the alteration of effector CD8+ T cell in phenotype, proliferation, and function. Immunohistochemistry (IHC) was used to investigate immune cell tumor-infiltration. Results: The established murine model reflects the typical features of human HCC. ABX oral administration significantly changed gut microbiota composition, particularly in abundance of P. aeruginosa and Muribaculaceae. ABX-mediated alteration of gut microbiota significantly retarded HCC progression. Interestingly, ABX treatment was found to prevent adoptively transferred tumor antigen-specific CD8+ T cell from tumor-induced immune tolerance, as higher frequency of effector CD8+ T cell was detected in ABX-treated tumor-bearing mice compared to that in control tumor-bearing mice. Correspondingly, ABX treatment resulted in the increase in expression of CD69 and enhanced production of IFN-γ in tumor antigen-specific CD8+ T cells relevant to that in control mice. Conclusion: Modulating microbiota with antibiotics is able to shape intrahepatic immune responses and therapeutically suppress tumor growth, providing a new option in HCC control. Citation Format: Xiaoqiang Qi, Ming Yang, Joseph Stenberg, Guangfu Li, Kevin Staveley-O’Carroll. Studying the impact of gut microbiome to hepatocellular cancer in an orthotopic murine model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4960.

Characterization and Inception of a Triterpenoid Astrakurkurol, as a Cytotoxic Molecule on Human Hepatocellular Carcinoma Cells, Hep3B.

Mushrooms are customary influential sources of pharmaceutically active metabolites. Usually lanostane-type triterpenoids from mushrooms had prospective for cancer disease treatments. Recently, a triterpenoid, astrakurkurol obtained from the fresh basidiocarps of the edible mushroom Astraeus hygrometricus, drew attention as a new cytotoxic therapeutic. The structural stability of this triterpenoid had been established with the amalgamation of density functional theory (DFT) calculations and study of single-crystal X-ray diffraction. To successfully manifest astrakurkurol as a potent cytotoxic therapeutics, a wide apprehension on the molecular and cellular mechanisms underlying their action is prerequisite. On this account, our study was directed to scrutinize the influence of this triterpenoid on human hepatocellular cancer cell model Hep3B. Encapsulating all experimental facts revealed that astrakurkurol had significantly decreased cell viability in a concentration-dependent manner. This effect was unveiled to be apoptosis, documented by DNA fragmentation, chromatin condensation, nuclear shrinkage, membrane blebing, and imbalance of cell cycle distribution. Astrakurkurol persuaded the expression of death receptor associated proteins (Fas), which triggered caspase-8 activation following tBid cleavage. Moreover, tBid mediated ROS generation, which triggered mitochondrial dysfunction and activated the mitochondrial apoptotic events. Astrakurkurol cytotoxicity was based on caspase-8-mediated intrinsic apoptotic pathway and was associated with inhibition at Akt and NF-κB pathway. Astrakurkurol had also inhibited the migration of Hep3B cells, indicating its antimigratory potential. These findings led us to introduce astrakurkurol as a feasible and natural source for a safer cytotoxic drug against hepatocellular carcinoma.

A multifunctional supramolecular vesicle based on complex of cystamine dihydrochloride capped pillar[5]arene and galactose derivative for targeted drug delivery.

Background: Supramolecular vesicles are a novel class of nanocarriers that have great potential in biomedicine.Methods: A multifunctional supramolecular vesicle (CAAP5G) based on the complex of CAAP5 and galactose derivative (G) assembled via host-guest interaction was constructed.Results: Using Human embryonic kidney T (293T) cells as experimental models, the cytotoxic effects of CAAP5G was investigated to 0–50 µmol/L for 24 h. Notably, the CAAP5G vesicles revealed low-toxicity to 293T cells, it was critical to designing drug nano-carriers. Simultaneously, we have evaluated doxorubicin hydrochloride (DOX)-loaded CAAP5G vesicles anticancer efficiency, where DOX-loaded CAAP5G vesicles and free DOX incubated with Human hepatocellular carcinoma cancer cell (HpeG2 cells) and 293T cells for 24 h, 48 h, 72 h. It turned out that CAAP5G vesicles encapsulated anticancer drug (DOX) could decrease DOX side-effect on 293T cells and increase DOX anticancer efficiency. More importantly, the cysteamine as an adjuvant chemotherapy drug was released from CAAP5G vesicles in HepG2 cells where a higher GSH concentration exists. The adjuvant chemotherapy efficiency was evaluated, where free DOX and DOX-loaded CAAP5G vesicles incubated with DOX-resistance HepG2 cells (HepG2-ADR cells) for 24, 48, 72 h, respectively.Conclusion: The results revealed that the DOX encapsulated by CAAP5G vesicles could enhance the cytotoxicity of DOX and provide insights for designing advanced nano-carriers toward adjuvant chemotherapies.

Tankyrase Promotes Aerobic Glycolysis and Proliferation of Ovarian Cancer through Activation of Wnt/β-Catenin Signaling

Tankyrase (TNKS) plays important roles in the malignancy of several cancers such as human lung tumor, breast cancer, and hepatocellular cancer. However, its exact functions and molecular mechanisms in ovarian cancer remain unclear. In this study, we found that TNKS was aberrantly overexpressed in human ovarian cancer tissues and associated with poor patient prognosis. TNKS inhibition or knockdown not only reduced ovarian cancer cell proliferation, colony formation, migration, invasion, and tumorigenic potential in nude mice but also enhanced the drug susceptibility of ovarian cancer cells through arresting cell cycle and inducing apoptosis. These phenotypic changes correlated with downregulation of targets (Cyclin D1, MDR, and MMP-9) of Wnt/β-catenin signaling. Furthermore, downregulation of TNKS suppressed the glucose uptake, lactate excretion, and cellular ATP levels and increased cellular O2 consumption rates. Molecular mechanism studies revealed that TNKS promoted aerobic glycolysis at least in part due to upregulation of pyruvate carboxylase (PC) via activation of Wnt/β-catenin/snail signaling. In agreement with these findings, expression of TNKS is positively associated with snail and PC in clinical ovarian cancer samples. Our findings identified TNKS as an oncogenic regulator of ovarian cancer cells proliferation that promotes aerobic glycolysis via activation of Wnt/β-catenin signaling, indicating that the TNKS might serve as a potential molecular target for clinical therapy of Wnt/β-catenin dependent ovarian cancer.

Expression of hUTP14a in non-small cell lung cancer

Human U three protein 14a (hUTP14a) facilitates tumorigenesis through promoting p53 and Rb degradation as well as enhancing c-Myc oncogenic activity. Moreover, hUTP14a expression is up-regulated in human hepatocellular cancer and colorectal cancer tissues. In this study, the expression of hUTP14a in non-small cell lung cancer (NSCLC) tissues was evaluated by immunohistochemistry staining (IHC). The relationship between hUTP14a expression levels and the clinical characteristics of the NSCLC patients were analyzed. Lung cancer tissues and the adjacent non-cancerous tissues were collected from 123 cases of NSCLC patients including 53 cases of squamous cell carcinoma (SCC) and 70 cases of adenocarcinoma (ADC), who had accepted surgical resection at Peking University Third Hospital from May 2003 to April 2006. The expression level of hUTP14a was determined by IHC in human NSCLC tissues and the adjacent non-cancerous tissues. The associations between hUTP14a expression and the clinical pathological variables including gender, age, tumor size, histological type, differentiation degree and clinical pathological stage were analyzed using the Pearson's χ2 test. The expression rate of hUTP14a in NSCLC tissues was significantly higher than that in the non-cancerous tissues (37.4% vs. 0, P<0.001). The expressions of hUTP14a in lung ADC and SCC were 48.6% and 20.6%, respectively. The expression rate of hUTP14a in both lung ADC and SCC was significantly higher than that in the adjacent non-cancerous tissues (P<0.001). In addition, the expression rate of hUTP14a in lung ADC was significantly higher than that in SCC (χ2=8.66, P=0.003). Furthermore, the expression rate of hUTP14a in the late pTNM stage of SCC was significantly higher than that in the early pTNM stage of SCC while hUTP14a expression level was not associated with pTNM stage of ADC. No correlation was found between hUTP14a expression and the other clinical pathologic features of the patients. Expression of hUTP14a was up-regulated in NSCLC tissues and was correlated with pTNM stage of SCC, suggesting that hUTP14a might possess a potential as a candidate marker for the early diagnosis screening of NSCLC.

Overexpression of Opa interacting protein5 increases the progression of liver cancer via BMPR2/JUN/CHEK1/RAC1 dysregulation.

Opa interacting protein 5 (OIP5) overexpression is associated with human carcinoma. However, its biological function, underlying mechanism and clinical significance in liver cancer remain unknown. In the present study, the effects of OIP5 expression on liver cancer, and the mechanisms regulating these effects, were investigated. OIP5 expression was measured in human hepatocellular carcinoma (HCC) tissues and liver cancer cell lines. The effect of OIP5 knockdown on tumorigenesis was also detected in nude mice, and differentially-expressed genes (DEGs) were identified and their biological functions were identified. The results indicated that OIP5 expression was significantly upregulated in HCC tissues and four liver cancer cell lines (P<0.01). Increased OIP5 protein expression significantly predicted reduced survival rate of patients with HCC (P<0.01). OIP5 knockdown resulted in the suppression of proliferation and colony forming abilities, cell cycle arrest at the G0/G1 or G2/M phases, and promotion of cell apoptosis. A total of 628 DEGs, including 87 upregulated and 541 downregulated genes, were identified following OIP5 knockdown. Functional enrichment analysis indicated that DEGs were involved in ‘RNA Post-Transcriptional Modification, Cancer and Organismal Injury and Abnormalities’. Finally, OIP5 knockdown in Huh7 cells dysregulated bone morphogenetic protein receptor type 2/JUN/checkpoint kinase 1/Rac family small GTPase 1 expression. In conclusion, the overall results demonstrated the involvement of OIP5 in the progression of liver cancer and its mechanism of action.

Antitumor activities of new iso(thio)cyanates and their nitrogen and sulphur heterocyclic phosphorus derivatives

Reaction of nucleophilic active phosphacumulenes with iso(thio)cyanates were performed. Novel heterocyclic four-membered or six-membered compounds containing sulfur and nitrogen nucleus namely, thietane, azetidine and thiazinane were prepared. Moreover, the thiocarbamoyl derivatives were produced from the reaction of the stable phosphonium ylides with the iso(thio)cyanate derivatives. Besides the cytotoxic activity of iso(thio)cyanate derivatives and some new organophosphorus compounds were determined, using MTT assay against human HepG2 hepatocellular carcinoma and MCF-7 adenocarcinoma breast cancer cell lines in comparison with the activity of the known anticancer drug 5-flurouracil as a reference drug. All the tested compounds showed concentration dependent increase in the growth inhibition percentage against HepG2 and MCF-7 cancer cell lines. Moreover, the synthesized thietanes and thiazinanes revealed anticancer activity with IC50 (20, 8.9, 12.7, 32.5 µg/mL) respectively against HepG2 and (20, 10.3, 20 and 20 µg/mL) respectively against MCF-7 cancer cell lines. The newly synthesized azetidines showed anticancer activity with IC50 (13.5 and 32.5 µg/mL) against HepG2 and IC50 (10 and 25.9 µg/mL) against MCF-7 cancer cell lines. Moreover, azetidinedione compound exhibited more potent activity than the azetidinone with both types of cancer cell lines. In addition the cytotoxic activity of the iso(thio)cyanates , and malonamic acid methyl ester compounds were also investigated. 4-Methoxyphenylisothiocyanate and methylisothiocyanate, with IC50 (25.9, 12.3 and 40, 20 µg/mL) respectively against HepG2 and MCF-7 cancer cells were reported. 1,2-Dichloro-4-isocyanatobenzene was simillar in potency to the known anticancer drug 5-flurouracil with IC50 (5.3 µg/mL) versus 5 µg/mL for 5-flurouracil against MCF-7. While, malonamic acid methyl ester compound had no effect on both types of cancer cell lines.

New Self-assembled Supramolecular Bowls as Potent Anticancer Agents for Human Hepatocellular Carcinoma

We report herein on the design, synthesis and biological activity of Ru-based self-assembled supramolecular bowls as a potent anticancer therapeutic in human hepatocellular cancer. The potent complex induces production of reactive oxygen species (ROS) by higher fatty acid β-oxidation and down-regulation of glucose transporter-mediated pyruvate dehydrogenase kinase 1 via reduced hypoxia-inducible factor 1α. Also, overexpressed acetyl-CoA activates the tricarboxylic acid cycle and the electron transport system and induces hypergeneration of ROS. Finally, ROS overexpressed through this pathway leads to apoptosis. Furthermore, we demonstrate that the naphthalene derived molecular bowl activates classical apoptosis via crosstalk between the extrinsic and intrinsic signal pathway. Our work into the mechanism of Ru-based self-assembled supramolecular bowls can provide valuable insight into the potential for use as a promising anticancer agent.

Synthesis, Structure Elucidation, and Biological Activities of Pyrazoles Against Human Lung and Hepatocellular Cancer

Two new series of pyrazoles and bipyrazoles were synthesized, and their structures were elucidated according to all data results from spectral and elemental analyses. The pathway of the formation of the new bipyrazole derivatives was discussed. Further, all the synthesized derivatives were screened for their antitumor activity on human lung (A‐549) and hepatocellular cancer (HepG‐2) cell lines. The results of antitumor screening have exhibited that several derivatives are more potent than the reference drug.

Evaluation of The Cytotoxicity, Cell Cycle Perturbations and Apoptotic Induction in Human Normal and Cancer Liver Cell Lines Exposed to Potassium Nitrate and Sodium Benzoate

This proposal investigates in vitro cytotoxic and apoptotic effects of two food preservatives commonly consumed in daily meals; potassium nitrate and sodium benzoate. The effect of these two preservatives on cell viability was tested on two different cell lines; normal liver cell line THLE2 and human hepatocellular carcinoma cancer cell line HepG2 using MTT assay. Although the effect was more significantly realized in HepG2 cell line, the viability of both cell lines was decreased by both of the tested compounds. Flow cytometric analysis of the HepG2 treated cells has revealed an increase in G2/M phase cell cycle arrest. Measurement of expression levels of three central genes; p53, bcl-2 and bax that play key roles in cell cycle and apoptosis were carried out in HepG2 using real time-PCR and western blot. In conclusion, both of the tested compounds have decreased the cell line viability and induced both cell cycle arrest and apopototic events indicating their high potential of being cytotoxic and genotoxic materials.