Esophageal Squamous Cell Proliferation Research Articles

PABPC1-induced stabilization of IFI27 mRNA promotes angiogenesis and malignant progression in esophageal squamous cell carcinoma through exosomal miRNA-21-5p

BackgroundEmerging evidence has demonstrated that RNA-binding protein dysregulation is involved in esophageal squamous cell carcinoma (ESCC) progression. However, the role of poly (A) binding protein cytoplasmic 1 (PABPC1) in ESCC is unclear. We therefore aimed to explore the functions and potential mechanisms of PABPC1 in ESCC progression.MethodsPABPC1 expression was characterized using immunohistochemistry and qRT-PCR in ESCC tissues and cell lines. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were used to detect histone acetylation in the promoter region of PABPC1. A series of in vitro and in vivo assays were further applied to elucidate the functions and underlying molecular mechanisms of PABPC1 in ESCC angiogenesis and malignant procession.ResultsPABPC1 expression was upregulated in ESCC tissues compared with in normal esophageal epithelial tissues. Elevated PABPC1 expression was correlated with tumor cell differentiation and poor prognosis in patients. Sp1 and p300 cooperated to increase the level of H2K37ac in the PABPC1 promoter. Functionally, PABPC1 overexpression enhanced esophageal squamous cell proliferation and invasion by activating the IFN/IFI27 signaling pathway. PABPC1 interacted with eIF4G to increase the stability of IFI27 mRNA by competing with RNA exosomes in ESCC. Furthermore, PABPC1/IFI27 could increase miR-21-5p expression to enable exosomal delivery of miR-21-5p to human umbilical vein endothelial cells to increase angiogenesis via inhibiting CXCL10.ConclusionPABPC1 plays a critical role in ESCC malignant progression by interacting with eIF4G to regulate IFI27 mRNA stability and promote angiogenesis via exosomal miR-21-5p/CXCL10. Taken together, our results suggest that PABPC1 is a promising therapeutic target for ESCC.

Effects of dietary zinc deficiency on esophageal squamous cell proliferation and the mechanisms involved.

BACKGROUNDDietary zinc deficiency has been shown to be associated with the development of esophageal cancer in humans, but the exact mechanism of action is not knownAIMTo observe the effects of dietary zinc deficiency on esophageal squamous cell proliferation. METHODSThirty C57BL/6 mice were randomly divided into three groups: A zinc-sufficient (ZS) group, zinc-deficient (ZD) group, and zinc-replenished (ZR) group. For weeks 1–10, zinc levels in the mice diets were 30.66–30.89 mg/kg in the ZS group and 0.66–0.89 mg/kg in the ZD and ZR groups. During weeks 10–12, the ZR group was switched to the ZS diet; the other two groups had no changes in their diets. Changes in body weight, serum, and esophageal tissue zinc concentrations were assessed as well as differences in the expression of proliferating cell nuclear antigen (PCNA), mitogen-activated protein kinase p38 (p38MAPK), nuclear factor kappa B (NF-κB) p105, NF-κB p65, and cyclooxygenase (COX)-2 proteins in the esophageal mucosa.RESULTSThe body weight and zinc concentration in the serum and esophageal mucosa were significantly lower in the ZD and ZR groups than in the ZS group (P < 0.05). In ZD mice, there was a marked proliferation of basal cells in the esophageal mucosa, resulting in a disturbance in the arrangement of basal cells in layers 2–4, a thickening of the squamous layer, and a significant increase in the expression of the above-mentioned five proteins involved in proliferation and inflammation in the esophageal mucosa. Two weeks after switching to the ZS diet, the serum zinc concentration in the ZR group increased, and the expression of PCNA, NF-κB p105, and COX-2 decreased, but the concentration of zinc in the esophageal mucosa and the structure of the esophageal mucosa did not display any significant changesCONCLUSIONThe ZD diet decreased the growth rate and promoted the proliferation of esophageal squamous cells in mice. The mechanism of proliferation was related to the induced overexpression of COX-2, P38, PCNA, and NF-κB (p105 and p65), and the ZR diet reduced the expression of PCNA, NF-κB p105, and COX-2, thereby reversing this process.​

Effect of Dietary Zinc Deficiency on Proliferation of Esophageal Squamous Cells in Mice and Related Mechanism

Effect of Dietary Zinc Deficiency on Proliferation of Esophageal Squamous Cells in Mice and Related Mechanism

MicroRNA (MiR)-301a-3p regulates the proliferation of esophageal squamous cells via targeting PTEN

ABSTRACT Human esophageal carcinoma (EC) is a common cancer, which leads to many deaths worldwide every year. Our study aimed to explore the mechanism of miR-301a-3p regulating the proliferation of esophageal squamous cell carcinoma (ESCC) cells. We had collected ESCC tissues and adjacent normal esophageal tissues from 47 patients. The relative levels of miR-301a-3p/U6 in ESCC tissues and cells were analyzed by real-time PCR. And we measured the relative protein levels of PTEN, BCL-2, BAX, and p-AKT/AKT by Western blot. Eca-109 cell proliferation was detected by MTT assay and colony formation. Compared with adjacent normal esophageal tissues, the relative level of miR-301a-3p/U6 was elevated in ESCC tissues. MiR-301a-3p could facilitate ESCC cell proliferation. And miR-301a-3p directly bind to PTEN 3ʹ-UTR and negatively regulated PTEN protein expression. Moreover, silencing PTEN could reversed inhibited proliferation of Eca-109 cells induced by miR-301a-3p inhibitor, while overexpression PTEN could reversed enhanced proliferation of Eca-109 cells induced by miR-301a-3p mimic. Taken together, miR-301a-3p promoted ESCC cell proliferation by supressing PTEN.

Relationship between expression of genes associated with retinoid interferon induced mortality-19, signal transducer and activators of transcription 3, and myeloid cell leukemia-1 with proliferation and apoptosis of esophageal squamous carcinoma cells

Objective To investigate the relationship between the expression of genes associated with retinoid interferon induced mortality-19 (GRIM-19), signal transducer and activators of transcription 3 (STAT3), and myeloid cell leukemia-1 (Mcl-1) with esophageal squamous carcinoma cells proliferation and apoptosis and the possible mechanism. Methods The protein expression of GRIM-19, STAT3, Mcl-1 and proliferation cell nuclear antigen (Ki-67) in 50 cases of esophageal squamous carcinoma tissues, 30 cases of atypical tissue adjacent to carcinoma and 50 cases of normal esophageal mucosa tissues was detected by immunohistochemical streptavidin avidin-peroxidase (SP) method. The mRNA expression of GRIM-19, STAT3, Mcl-1 and Ki-67 in 50 cases of esophageal squamous carcinoma tissues, 30 cases of atypical tissue adjacent to carcinoma and 50 cases of normal esophageal mucosa tissues was detected by in situ hybridization. TdT-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) method was applied to examine apoptosis of esophageal squamous carcinoma cells. Results The positive expression rate of GRIM-19 protein and mRNA in esophageal squamous carcinoma tissues was significantly lower than that in atypical hyperplasia tissue adjacent to carcinoma and normal esophageal mucosa (protein: χ2=12.130, P=0.000; mRNA: χ2=11.830, P=0.000). The positive expression rate of STAT3 and Mcl-1 protein and mRNA in esophageal squamous carcinoma tissues was significantly higher than that in atypical hyperplasia tissue adjacent to carcinoma and normal esophageal mucosa (STAT3 protein: χ2=4.910, P=0.000, STAT3 mRNA: χ2=5.410, P=0.000; Mcl-1 protein: χ2=4.970, P=0.000, Mcl-1 mRNA: χ2=5.370, P=0.000). The apoptosis of tumor cells in esophageal squamous carcinoma tissues was associated with the positive expression rate of GRIM-19, STAT3 and Mcl-1. The apoptosis index (AI) in the group of GRIM-19 positive expression was higher than that in the group of GRIM-19 negative expression (F=18.355, P=0.048). The AI in the group of STAT3 positive expression was lower than that in the group of STAT3 negative expression (F=19.003, P=0.036). The AI in the group of Mcl-1 positive expression was lower than that in the group of Mcl-1 negative expression (F=18.225, P=0.044). Conclusion The down-regulation of GRIM-19, and up-regulation of STAT3 and Mcl-1 may promote esophageal squamous cancer cell proliferation, and inhibit esophageal squamous cancer cell apoptosis. Key words: Genes associated with retinoid interferon induced mortality-19; Signal transducer and activators of transcription 3; Myeloid cell leukemia-1; Esophageal squamous carcinoma cells; Apoptosis

Effect of PC cell-derived growth factor RNA interference on biological behavior of esophageal squamous carcinoma cells

To investigate the effect of PC cell-derived growth factor (PCDGF) RNA interference on esophageal squamous carcinoma cells Eca-109 in vitro. The PCDGF-shRNA expression vector was transfected into the Eca-109 cells by liposome. After transfection, the mRNA and protein expressions of PCDGF were detected by RT-PCR and Western-blot respectively. Cell Counting Kit-8 (CCK-8) assay and Boyden chamber method were performed to measure the cell proliferation and invasion ability respectively. The expression levels of PCDGF mRNA and protein were both decreased in Eca-109 cells transfected with PCDGF-shRNA expression vector (transfection group). Twenty-four, 48 and 72 h after transfection, the cells proliferation in the transfection group was inhibited, and the inhibition rate was 20.4%, 21.1% and 20.9% respectively. The cell proliferation activity in the transfection group was significantly lower than that in the non-transfection group, liposome group and negative vector group (all P<0.05). The number of cell migration in the non-transfection group,negative vector group, liposome group and transfection group was 118.8±12.0, 100.8±9.0, 114.3±4.7, and 53.5±16.3 respectively. The differences were statistically significant between the transfection group and the other 3 groups (all P<0.05). PCDGF RNA interference can inhibit the proliferation and invasion abilities of esophageal squamous carcinoma cells in vitro. PCDGF gene may be the new target of gene therapy.

Expression of NDRG2 in human lung cancer and its correlation with prognosis

We had reported that N-myc downstream–regulated gene (NDRG2) regulates colorectal cancer, breast cancer, clear cell renal cell carcinoma, pancreatic cancer, thyroid cancer and esophageal squamous cell proliferation, development, and apoptosis. The goal of this study was to determine the expression pattern of NDRG2 in human lung cancer and its correlation with prognosis. Immunohistochemistry, RT-PCR and western blot were used to explore the expression of NDRG2 in 185 human lung cancer patients. The correlation of NDRG2 expression with patients’ survival rate was assessed by Kaplan–Meier and Cox regression. Results showed that the expression level of NDRG2 was decreased in human lung cancer tissues, and NDRG2 was positively correlated with depth of invasion (P = 0.038), vascular invasion (P = 0.036), tumor grade (P = 0.039), and tumor size (P = 0.026). Both RT-PCR and Western blots demonstrated that NDRG2 mRNA and protein levels were lower in lung cancer compared to the adjacent normal tissue from the same individual, and NDRG2 level was negatively correlated with UICC stage. Additionally, survival time of lung cancer patients with high expression of NDRG2 was longer than those with low expression during the 5-year follow-up period (P = 0.001). Meanwhile, COX regression analysis indicated that low expression of NDRG2, ≥pT3, pM1, ≥pN1 and vascular invasion were independent, poor prognostic factors of lung cancer patients. These data showed that NDRG2 may play an important role in human lung cancer tumourigenesis, and NDRG2 might serve as a novel prognostic marker in human lung cancer.

Effects of EZH2-siRNA on proliferation and invasion of human esophageal squamous carcinoma cells

Objective To investigate the influence of EZH2 gene down-regulation by RNA interference (RNAi) on the proliferation and invasion of human esophageal squamous carcinoma cell line Ecal09.Methods Two small interfering RNAs (siRNA) targeting EZH2 were transfected into Eca109 cells by using lipofectamine 2000.The proliferation capcity of Eca109 cells was determined by calculating the cell population doubling time,the cell cycle,the cell cloning and the percentage of apoptotic cells.The invasive ability of Eca109 cells was detemined by Transwell assay.Results In vitro experiment showed that Eca109 cells transfected with EZH2-siRNA had significantly longer cell population doubling time [ ( 26.7 ± 0.9) h vs ( 22.6 ± 0.8 ) h,P ＜ 0.01 ],lower percentage of cell cloning [ ( 39.1 ± 2.3 ) ％ vs(65.1±3.0)％,P＜0.01],higher cell apoptotic rate [(9.51±0.19)％ vs (1.11 ±0.08)％,P＜0.01 ],and lower cell invasion rate [ (35.0 ± 0.4) ％ vs (51.8 ± 0.7) ％,P ＜ 0.01 ] than control groups.However,there was no significant diference in the cell cycle at S phase between EZH-2-siRNA group and control group [(32.1 ±0.7)％ vs (31.9 ±0.4)％,P＞0.05].Conclusion EZH2 siRNA can reduce the proliferation and invasion of human esophageal squamous carcinoma cells,which suggests that EZH2 siRNA might be a potential gene therapertic target of human esophageal squamous carcinoma. Key words: Esophageal squamous carcinoma; EZH2; RNA interference; Proliferation; Invasion

Telomere Dysfunction and DNA Damage Checkpoints in Diseases and Cancer of the Gastrointestinal Tract

Telomere Dysfunction and DNA Damage Checkpoints in Diseases and Cancer of the Gastrointestinal Tract

Small interfering RNA suppression of transducer and activator of transcription 3 (STAT3) signaling pathway: inhibitory effect on proliferation of human esophageal squamous carcinoma cells

To investigate the efficiency of blockage of constitutively activated STAT3 signaling by small interfering RNA (siRNA), and to explore the inhibitory effects on the proliferation of human esophageal squamous carcinoma cells (EC9706 and Eca109). EC9706 and Eca109 were transfected with chemical synthesized STAT3 siRNA (100 nmol/L). RT-PCR and Western blot were used to detect STAT3 mRNA and protein expression, including phosphorylated-STAT3 (p-STAT3) before and after the transfection respectively. The changes of DNA-binding activity and cell proliferation were evaluated by electrophoretic mobility gel shift assay and MTT, respectively. Stages of cell cycle were determined by flow cytometry. Expression levels of STAT3 mRNA and STAT3, p-STAT3 proteins were progressively inhibited by STAT3 siRNA at various time points after transfection. STAT3-DNA-binding activity was suppressed after transfection evidenced by electrophoretic mobility gel shift assay. The cell cycle was arrested at G(0)/G(1) phase along with a significant inhibition of cell proliferation after STAT3 siRNA treatment. STAT3 siRNA specifically and efficiently blocks the constitutively activated STAT3 signaling pathway in human esophageal squamous carcinoma cells, resulting in cell cycle arrest and proliferation inhibition.