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Objective: To explore the key deubiquitinating enzymes that maintain the stemness of liver cancer stem cells and provide new ideas for targeted liver cancer therapy. Methods: The high-throughput CRISPR screening technology was used to screen the deubiquitinating enzymes that maintain the stemness of liver cancer stem cells. RT-qPCR and Western blot were used to analyze gene expression levels. Stemness of liver cancer cells was detected by spheroid-formation and soft agar colony formation assays. Tumor growth in nude mice was detected by subcutaneous tumor-bearing experiments. Bioinformatics and clinical samples were examined for the clinical significance of target genes. Results: MINDY1 was highly expressed in liver cancer stem cells. The expression of stem markers, the self-renewal ability of cells, and the growth of transplanted tumors were significantly reduced and inhibited after knocking out MINDY1, and its mechanism of action may be related to the regulation of the Wnt signaling pathway. The expression level of MINDY1 was higher in liver cancer tissues than that in adjacent tumors, which was closely related to tumor progression, and its high expression was an independent risk factor for a poor prognosis of liver cancer. Conclusion: The deubiquitinating enzyme MINDY1 promotes stemness in liver cancer cells and is one of the independent predictors of poor prognosis in liver cancer.
Subject(s)
Animals , Mice , Cell Line, Tumor , Mice, Nude , Liver Neoplasms/pathology , Prognosis , Deubiquitinating Enzymes/metabolism , Neoplastic Stem Cells/pathology , Gene Expression Regulation, NeoplasticABSTRACT
Objective To observe the effects of amarogentinon liver cancer stem cells (LCSCs) after insufficient thermal ablation and its mechanism. Methods A insufficient thermal ablation model of HepG2 cells was established by water bath method.The percentage of CD133-positive LCSCs and the mRNA and protein levels of CD133 were detected by flow cytometry, qRT-PCR and Western blot.The insufficient thermal ablation model of HepG2 cells was treated with variable doses of amarogentin for 24 h; the percentage of CD133-positive LCSCs, the proliferation and apoptosis of liver cancer cells, and the mRNA and protein levels of CD133, TBC1D15, and p53were detected by flow cytometry, qRT-PCR and Western blot. Results The percentage of CD133-positive HepG2 cells and the mRNA and protein levels of CD133 and TBC1D15in the insufficient thermal ablation model were significantly higher than those in the normal HepG2 cells.Amarogentin then markedly decreased the percentage of CD133-positive LCSCs, the proliferation rate of HepG2 cells, and the mRNA and protein levels of CD133 and TBC1D15 in the insufficient thermal ablationresidual model (all P < 0.05);inversely, the apoptosis rate of HepG2 cells and the phosphorylated levels of p53 in the insufficient thermal ablation model were significantly increased (all P < 0.05). Conclusion Amarogentin could reduce the proportion of LCSCs after insufficient thermal ablation, inhibit the proliferation, and promote the apoptosis of LCSCs, which maybe associated with increasing the phosphorylation of p53 and inhibiting the expression of TBC1D15.
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Primary liver cancer (PLC) is an aggressive tumor and prone to metastasize and recur. According to pathological features, PLC are mainly categorized into hepatocellular carcinoma, intrahepatic cholangiocarcinoma, mixed hepatocellular cholangiocarcinoma, and fibrolamelic hepatocellular carcinoma, etc. At present, surgical resection, radiotherapy and chemotherapy are still the main treatments for PLC, but the specificities are poor and the clinical effects are limited with a 5-year overall survival rate of 18%. Liver cancer stem cells (LCSCs) are a specific cell subset existing in liver cancer tissues. They harbor the capabilities of self-renewal and strong tumorigenicity, driving tumor initiation, metastasis, drug resistance and recurrence of PLC. Therefore, the identification of molecular markers and the illustration of mechanisms for stemness maintenance of LCSCs can not only reveal the molecular mechanisms of PLC tumorigenesis, but also lay a theoretical foundation for the molecular classification, prognosis evaluation and targeted therapy of PLC. The latest research showed that the combination of 5-fluorouracil and CD13 inhibitors could inhibit the proliferation of CD13+ LCSCs, thereby reducing overall tumor burden. Taken together, LCSCs could be the promising therapeutic targets of PLC in the future. This review summarizes the latest progress in molecular markers, mechanisms for stemness maintenance and targeted therapies of LCSCs.
Subject(s)
Humans , Carcinoma, Hepatocellular/genetics , Liver Neoplasms/genetics , Neoplastic Stem Cells , PrognosisABSTRACT
With the in-depth study of alpha-fetoprotein (AFP) and lymphocyte immune regulation, tumor suppressor protein factor and stem cell factor, it is found that AFP is involved in tumorigenesis, growth, metastasis and maintenance of liver cancer cell stem cell function. AFP inhibits natural killer cell activity by inhibiting lymphatic system to protect the liver cancer cells from immune surveillance; AFP can also inhibit the activity of tumor suppressor genes such as caspase-3, pten, p53 and tumor suppressor proteins; AFP can promote the formation of liver cancer stem cells and maintains stability through activating the expressions of other cancer stem cells factors. This paper reviews the biological functions of AFP by regulating hepatocellular carcinoma cells to evade immune surveillance, inhibiting apoptosis and maintaining the generation of liver cancer stem cells.
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Objective To determine the effect of COMMD7 inhibition on invasion and migration in liver cancer stem cells (LCSCs),and investigate the possible mechanism.Methods After LCSCs were infected by shRNA lentiviral vectors of COMMD7,adhesion assay and Transwell assay were used to detect the invasion and migration,and phalloidin staining was employed to observe the morphological changes.Western blotting was adopted to measure the expression of E-cadherin,N-cadherin and Vimentin.Results COMMD7 knockdown significantly inhibited the invasion and migration of LCSCs.The relative cell quantity of adhesion was 1.00 ± 0.12 and 2.35 ± 0.20 respectively in control cells and infected cells,suggesting there were significantly more adhesive cells in the infected group (P ＜ 0.05).The relative cell quantity per visual field of migration was 1.00 ±0.04 and 0.24±0.03,and that of invasion was 1.00 ±0.05 and 0.24 ±0.04 respectively in the control cells and infected cells,and there were significantly less invasive and migrated cells in the infected group (P ＜0.05).What's more,COMMD7 knockdown also induced some morphological changes of cells corresponding to the weakened abilities of migration and invasion.All the changes above were associated with up-regulation of E-cadherin (P ＜ 0.05) and down-regulation of N-cadherin and Vimentin (P ＜0.05),the molecules related to mesenchymal-epithelial transition (MET).Conclusion COMMD7 knockdown inhibits the invasion and migration in LCSCs,which may be through its regulation on the MET course.
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Over several decades, a hierarchical cancer stem cell (CSC) model has been established in development of solid cancers, including hepatocellular carcinoma(HCC). In terms of this concept, HCCs originate from liver CSCs. Clinically HCCs show a wide range of manifestations from slow growth to very aggressive metastasis. One of the reasons may be that liver CSCs originate from different cells. This review describes the basic concept of CSCs and the cellular origin of liver CSCs.
Subject(s)
Carcinoma, Hepatocellular , Liver , Liver Neoplasms , Neoplasm Metastasis , Neoplastic Stem Cells , Stem CellsABSTRACT
Liver cancer stem cells (LCSC)play the critical role in hepatocellular carcinoma development and maintenance. They are generally dormant or slowly cycling tumor cells that have the ability to reconstitute tumors. LCSC associate closely with tumor resistance to chemo/radiation therapy , tumor relapse and metastasis, and can be identified and separated with some special surface markers from hepatocellular carcinoma, such as CD133, CD90, CD44, CD24 and EpCAM to investigate the biological behaviors of them. Early studies showed that these markers can be regarded as special surface markers of liver cancer stem cells. Recent studies found that aminopeptidase N (APN,CD13+)cells in hepatocellular carcinoma have biological characteristics of stem cells and demonstrated that CD13 is a marker for semiquiescent CSC in human liver cancer cell lines and clinical samples and that targeting these cells might provide a way to treat this disease. In this review,we introduce the structure and the main function of CD13,liver cancer stem cells source and identification,CD13 + CSC in hepatocellular carcinoma and combination therapy in the treatment of liver cancer.
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Liver cancer stem cells (LCSC) play the critical role in hepatocellular carcinoma development and maintenance. They are generally dormant or slowly cycling tumor cells that have the ability to reconstitute tumors. LCSC associate closely with tumor resistance to chemo/radiation therapy, tumor relapse and metastasis, and can be identified and separated with some special surface markers from hepatocellular carcinoma, such as CD133, CD90, CD44, CD24 and EpCAM to investigate the biological behaviors of them. Early studies showed that these markers can be regarded as special surface markers of liver cancer stem cells. Recent studies found that aminopeptidase N (APN, CD13+) cells in hepatocellular carcinoma have biological characteristics of stem cells and demonstrated that CD13 is a marker for semiquiescent CSC in human liver cancer cell lines and clinical samples and that targeting these cells might provide a way to treat this disease. In this review, we introduce the structure and the main function of CD13, liver cancer stem cells source and identification, CD13+ CSC in hepatocellular carcinoma and combination therapy in the treatment of liver cancer.
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The theory of cancer stem cells proposed that recurrence and metastasis of liver cancer are closely related to liver cancer stem cells.The traditional surgical treatment of liver cancer simply kills those rapidly proliferating tunor cells instead of eliminating hepatic cancer stem cells which play decisive role in recurrence and metastasis.As far as people have concerned,there are many signaling pathways and regulatory factors modulating liver cancer stem cells,such as the Wnt/β-catenin pathway,TGF-β pathway,Notch pathway,Hedgehog pathway,hepatitis B virus,as well as epigenetics.Further studies on regulatory mechanisms of liver cancer stem cells are of great significance,in hope of providing new evidence for treatment and prevention of recurrence and metastasis of liver cancer.
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Objective To study aminopeptidase N/CD13 expression in hepatocellular carcinoma and its relationship with clinical data and proguosis in patients with hepatocellular carcinoma.Methods The immunohistochemical SP method was used to detect the CD13 monoclonal antibody in 40 cases of hepatocellular carcinoma,10 cases of corresponding para-carcinoma and 10 cases of cavernous hemangioma.Results Forty cases ( 100％ ) hepatocellular carcinoma tissues were seen varying degrees of CD13 expression,3 (30％) corresponding para-carcinoma tissues were weakly positive,and 10 cases of cavernous hemangioma with no expression.The expression rate of CD13 was not significantly correlated with the patient gender,age,serum AFP value,HbsAg,differentiation,CHILD grade and TNM stage (P＞0.05).But the expression of CD13 was closely related with the patient serum AFP value,HbsAg,differentiation ( P ＜ 0.05 ).By the survival function graph we could find the expression rate was negativly correlated with survival in patients,but the expression was not significantly correlated with tumor relapse.Conclusion CD13 can be used as the surface marker of liver cancer stem cells,and it is expected to become an effective indicator of prognosis in patients with hepatocellular carcinoma.
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The hypothesis that tumor comes from stem cells has been demonstrated in various human tumors.Cancer is not only a genetic disease but also a stem cell disease. It is a key to regeneration, mutations and recurrence of tumors that gene mutations affect stem cells, and then stem cells mutate to cancer stem cells. In an effort to review the evidence that liver cancer stem cells exist, two fundamental questions must be addressed. First, do hepatocellular carcinomas(HCC)arise from liver stem cells? Second, do HCCs contain cells that possess properties of cancer stem cells?More recently, there is a hypothesis that HCC arise from maturation arrest of liver stem cells. Analysis of the cells in HCC supports the presence of cells with stem-cell properties(ie, immortality, transplantability, and resistance to therapy). However, definitive markers for these putative cancer stem cells have not yet been found and a liver cancer stem cell has not been isolated.
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Delta-like ligand 4(Dll4), one ligand of Notch receptors, is highly expressed in tumour vessels and is negative regulator in tumour angiogenesis. Blocking Dll4/Notch sinaling could promote tumour angiogenesis, whereas due to poor perrusion of newly forming vessels, tumour growth is inhibited, which is effective even to the tumour patients unresponsive to blocking vascular endothelial growth factor pathway. So, Dll4 is becoming a new promising target in anti-tumour theraties.