Esophageal Cancer Cell Lines TE-1 Research Articles

Study on Correlation and Mechanism of Procollagen Lysine 2 Oxoglutarate 5 Dioxygenase on Stem Cell Characteristics and Vasculogenic Mimicry Formation in Esophageal Squamous Cell Carcinoma

In this study, we investigated the significance of the stem cell transcription factor PLOD2 in esophageal cancer to enhance our understanding of its occurrence, development, recurrence, metastasis, and potential treatment. We conducted the following experiments: (1) Immunohistochemistry revealed elevated PLOD2 protein levels in esophageal carcinoma compared to adjacent tissues, with higher levels associated with advanced disease stages and lower differentiation. (2) Immunofluorescence demonstrated increased PLOD2 expression in esophageal cancer cell lines TE-1 and Eca- 109, suggesting a connection to cell differentiation. (3) We successfully transfected fluorescently labeled PLOD2 siRNA into cells, achieving a transfection rate of 67.57% and silencing efficiency exceeding 80%. (4) Following transfection, we observed a decreased proliferation rate in PLOD2 siRNA-treated cells, indicating that PLOD2 reduction can slow esophageal cancer growth. These findings emphasize that PLOD2 plays a crucial role in esophageal cancer pathogenesis, particularly in low cell differentiation maintenance. It could serve as a promising target for inducing differentiation in esophageal cancer and evaluating its malignancy. The high PLOD2 expression in esophageal cancer cell lines suggests the presence of tumor stem cells. Moreover, reducing PLOD2 through RNAi technology slows cell proliferation, suggesting that inhibiting PLOD2 may offer a potential therapeutic approach for esophageal cancer treatment.

Alkylation repair homolog 3-regulated esophageal squamous cell carcinoma associated long non-coding RNA 1 is required for maintaining the stemness of esophageal cancer

N1-methyladenosine (m1A) RNA modification represents one of the essential post-transcriptional modifications in gene expression regulation. Long non-coding RNAs (lncRNAs) are involved in the development of malignant tumors, including esophageal cancer (ESCA). However, whether m1A can regulate that lncRNA in cancer cells remains unclear. ESCA cell lines TE1 and KYSE70 were used for functional experiments. The mRNA and protein levels were detected by quantitative reverse transcription polymerase chain reaction and Western blot, respectively. Colony formation and tumor sphere formation assays were used for evaluating ESCA stemness. The m1A modification on esophageal squamous cell carcinoma associated long non-coding RNA 1 (ESCCAL-1) transcript was examined by methylated RNA immunoprecipitation. In this study, we report that RNA m1A demethylase alkylation repair homolog 3 (ALKBH3)-mediated ESCCAL-1 is implicated in maintaining stem cell-like properties of ESCA. Clinically, ESCCAL-1 was up-regulated in ESCA and positively correlated with tumor stage. In addition, patients with higher ESCCAL-1 expression in tumors had shorter median survival. Functionally, the knockdown of ESCCAL-1 attenuated the stemness of ESCA cells as indicated by decreased sphere formation and colony formation capacities, while overexpression of ESCCAL-1 elicits the opposite biological effects. Moreover, ESCCAL-1 manipulation positively regulated both mRNA and protein levels of KLF4 and CD44, two stemness-related markers. Mechanistically, ALKBH3 upregulated ESCCAL-1 expression in an m1A demethylation-dependent manner. Notably, the downregulation of ALKBH3 mimicked the effects of ESCCAL-1 deficiency on ESCA stemness, and this phenomenon is significantly reversed by the enforced expression of ESCCAL-1. Our results revealed the role of m1A-mediated ESCCAL-1 in ESCA self-renewal, which expands the understanding of lncRNA post-transcriptional modification in cancer development.

Poly(A)-Specific Ribonuclease (PARN) Promotes the Proliferation, Migration and Metastasis of Esophageal Cancer

Background: Esophageal cancer leads to the majority of cancer deaths in the world. Surgery, radiotherapy and chemotherapy are the main treatment methods, but due to its high recurrence and metastasis rate, the overall prognosis is poor. Poly(A)-specific ribonuclease (PARN) is a multifunctional enzyme molecule, which is involved in the occurrence and development of tumors. This study explores the relationship between PARN and the proliferation, metastasis and prognosis of esophageal cancer. Methods: The esophageal cancer tissues and adjacent normal tissues were collected from 91 patients with esophageal cancer after surgery. The expression level of PARN was detected by immunohistochemistry (IHC) using tissue microarray (TMA). PARN expression and its relationships with clinicopathological characteristics and prognosis was detected. In addition, stable PARN knockdown esophageal cancer cell lines TE-1 and Eca-109 were used to study its effect on the phenotype and behavior of tumor cells. Meanwhile, the effect of PARN silencing on tumor growth in vivo was verified by xenotransplantation model in nude mice. Finally, we examined possible downstream signaling mechanisms of PARN in esophageal cancer. Findings: PARN showed high expression in esophageal carcinoma tissues and its expression was significantly correlated with lymphatic metastasis and prognosis of patients. In the culture of PARN knockdown esophageal cancer cell lines, the growth and proliferation were significantly inhibited, and the invasion and migration ability of tumor cells were also significantly decreased. In addition, inhibition of PARN suppressed tumor growth and increased survival time in xenograft murine models. In the PARN knockdown group, BIM, IGFBP-5 and p21 were significantly increased, while the expression levels of anti-apoptotic proteins Survivin and STNF-R1 were significantly down-regulated. In addition, the expression levels of Akt, p-Akt, PIK3CA and CCND1 downstream of the signaling pathway regulating esophageal cancer progression were significantly down-regulated. Interpretation: These results suggest a previously undetected role of PARN as a suppressor of tumor development and its associations of prognosis in esophageal cancer through anti-apoptotic effect. Funding Information: None. Declaration of Interests: The authors declare no conflicts of interests. Ethics Approval Statement: Ethical approval for the study was approved by the Research Ethics Committee of the Second Affiliated Hospital of Guangxi Medical University and Zhujiang Hospital Affiliated to Southern Medical University, and written patient consent was obtained from all patients. Nude mice xenograft model - This study was approved by the Guangxi Medical University Ethics Committee.

MiR-502 mediates esophageal cancer cell TE1 proliferation by promoting AKT phosphorylation

Esophageal cancer is one of the most common cancers in the world and esophageal squamous cell carcinoma is one of the two main types in esophageal cancer. MicroRNA is a small non-coding RNA molecule functions in many different cancers including esophageal cancer. We found miR-502 was up-regulated in esophageal tissues, which indicated miRNA-502 may play important roles in esophageal cancer. In this study, we used esophageal cancer cell line TE1 as an in vitro model for investigating the role of miR-502 in promoting the proliferation of the cancer cells. We found that overexpressing miR-502 in TE1 cells promoted the proliferation and inhibited the apoptosis induced by dox. Down-regulating miR-502 made the opposite phenomenon. Furthermore, western blot showed that miR-502 enhanced the phosphorylation levels of AKT pathways, which may be the mechanism of the overgrowth for esophageal cancer cell. Our data provide the evidence of a role for miR-502 in the regulation the proliferation of esophageal cancer cell through promoting the phosphorylation of AKT signaling. Due to its ability to promote the overgrowth of esophageal cancer cell, miR-502 may be a novel target for esophageal cancer therapeutic.

Effects of diphyllin as a novel V-ATPase inhibitor on TE-1 and ECA-109 cells.

Diphyllin is a natural component of traditional Chinese medicine, which effectively inhibits V-ATPase activity and affects the progression of cancer. However, few studies have been conducted on esophageal cancer, and the mechanisms remain to be elucidated. The present study revealedthat diphyllin inhibited proliferation and induced S arrest in esophageal cancer cell lines TE-1 and ECA-109. Further experiments revealed that diphyllin inhibited V-ATPase activity and decreased the mRNA expression of mammalian target of rapamycin complex 1 (mTORC1), hypoxia-inducible factor-1α (HIF-1α), and vascular endothelial growth factor (VEGF). The present study also revealed that diphyllin inhibited proliferation and reduced the formation of new blood vessels. Diphyllin inhibited blood metastasis by regulating the mTORC1/HIF-1α-/VEGF pathway, therefore it could be considered as a new V-ATPase inhibitor to treat esophageal cancer.

Effects of siRNA-Mediated Knockdown of HDAC1 on the Biological Behavior of Esophageal Carcinoma Cell Lines.

BackgroundHDAC1 has been shown to be closely associated with the occurrence of tumors. We aimed to investigate the effects of siRNA-mediated HDAC1 knockdown on the biological behavior of esophageal carcinoma cell lines.Material/MethodsHDAC1 expression in esophageal cancer cell lines TE-1, Eca109, and EC9706 was compared by Western blot analysis. These cells were transfected with siRNA-HDAC1 and cell proliferation was evaluated by MTT assay to select the optimum cell line for subsequent experiments. The effects of siRNA-HDAC1 on the migration and invasion of the selected cell line were assessed by transwell assay. The expression of cell cycle-related proteins cyclinD1, p21 and p27, and epithelial-mesenchymal transition (EMT)-related protein zonula occludens-1 (ZO-1), E-cadherin and vimentin was determined by Western blot analysis.ResultsHDAC1 expression in TE-1, Eca109 and EC9706 cells was significantly higher compared with normal esophageal cell line HEEC (P<0.01). MTT assay, Western blot and RT-PCR analyses demonstrated that the inhibitory effects of siRNA on HDAC1 expression and cell viability in TE-1 cells were the highest among all cell lines, which was therefore used in subsequent experiments. After TE-1 cells were transfected with siRNA-HDAC1, their migration and invasion were significantly lower compared with the controls (P<0.01). CyclinD1 and vimentin expression was significantly lower compared with the controls (P<0.01), whereas the expression of p21, p27, ZO-1 and E-cadherin was significantly higher (P<0.01).ConclusionsThe siRNA-mediated HDAC1 knockdown significantly inhibited the proliferation, migration and invasion of TE-1 cells probably by regulating the expression of cell cycle- and EMT-related proteins.

Raf kinase inhibitor protein inhibits esophageal cancer cell invasion through downregulation of matrix metalloproteinase expression

Esophageal cancer is the eighth most common malignant tumor in the world and is a common cause of tumor-related death. The development of esophageal cancer is a complex process involving many pathogenetic factors, multiple stages and accumulation of multiple gene mutations and interactions. This study aimed to investigate the effects of Raf kinase inhibitor protein (RKIP) on the proliferation, apoptosis and invasion of TE-1 esophageal cancer cells. Surgical specimens from esophageal cancer patients were classified into esophageal cancer tissues, tumor-adjacent tissues and normal esophageal tissues. The tissues were fixed in 4% paraformaldehyde solution for hematoxylin and eosin and immunohistochemical staining. RKIP expression in esophageal tissues was detected by immunohistochemical staining. The esophageal cancer cell line TE-1 was exposed to four different viruses: RKIP-RNAi-AD, NC-RNAi-GFP-AD, RKIP-AD and GFP-AD. Cell proliferation was detected by MTT assay and cell apoptosis was detected by flow cytometry. Cell invasion was determined by a Transwell coated with Matrigel. RKIP, phospho-RKIP, Raf-1, phospho-Raf-1, ERK1/2, phospho-ERK1/2, GRK-2 and GAPDH expression was assayed by western blotting. LIN28 and MMP-14 mRNA was assayed by qPCR. The results showed that RKIP expression was reduced in esophageal cancer tissues in comparison with expression in normal esophageal epithelium tissues and tumor-adjacent tissues. Reduced RKIP expression was associated with lymph node or distant metastasis in esophageal cancer. RKIP inhibited the invasive and metastatic abilities of esophageal cancer cell line TE-1 by downregulating mRNA expression of LIN28 and MMP-14. RKIP had no effect on the MAPK signaling pathway in the esophageal cancer cell line TE-1, but was involved in the G protein-coupled signaling pathway. Our findings clearly demonstrate that RKIP inhibits esophageal cancer cell invasion by downregulating the expression of GRK-2, LIN28 and MMP-14.

Effect of Nitric Oxide on Esophageal Cancer Cell Line TE-1

To investigate the radiosensitizing effect of nitric oxide (NO) combined with radiation on esophageal cancer cell line TE-1. Methyl thiazolyl tetrazolium (MTT) assay was used to assess the effects of NO and radiation on TE-1 cells regarding inhibition of cell proliferation. Flow cytometry was used to examine the effect of NO and radiation on cell apoptosis and cycle. Reverse transcription polymerase chine reaction and Western blot were used to evaluete the effect of NO on mRNA and protein expression of manganese superoxide dismutase (MnSOD). NO inhibited the proliferation of TE-1 cells while significantly enhancing their radiosensitivity. The application of NO combined with radiation significantly increased the apoptosis rate and G2/M phase proportion of TE-1 cells, with substantial decreases in the MnSOD mRNA and protein expression levels. NO reduces the MnSOD mRNA and protein expression levels by affecting TE-1 cell cycle, further inhibiting the apoptosis of esophageal cancer cells and enhancing the killing effect of radiation on esophageal cancer cells.

Study on effect of BSO on esophageal cancer cell line TE-1

Objective The aim of the study was to investigate the sensitizing effect of buthionine sulfoximine (BSO) and radiation on esophageal cancer cell line TE-1.

The Combination of Olaparib and Camptothecin Effectively Sensitize Radiation

anticancer therapy, and radiation-induced autophagy contributes to resistance of cancer cells to ionizing radiation (IR). Our study is to examine whether autophagy inhibitor chloroquine (CQ) can sensitize esophageal cancer cell line TE-1 to IR. Materials/Methods: Cell viability of TE-1 cells treated with CQ was measured by using MTT assay. Formation and volume of acidic vesicular organelles (AVOs) in living cells after IR alone or IR combined with CQ were monitored by fluorescence staining with Lyso-Tracker Red DND-99/ Hoechst 33258 according to the cytoplasmic fractional volume occupied by AVO. The expression of autophagy-related protein LC3-I and LC3-II were determined by SDS-PAGE and Western blotting. The radiosensitivity of CQ at its IC10 concentration on TE-1 cells and survival TE-1 cells preirradiated with 2 Gy in the treatment with CQ before or after exposure to Co g-radiation was evaluated by clonogenic forming assay, and the radiation biology parameters were estimated from the curve fitted by the single-hit multi-target model [SF Z 1-(-e)]. Results: CQ showed dose-dependent inhibition of TE-1 cell growth and the values of IC50 and IC10 were 72.3 mmol/L and 15.4 mmol/L, respectively. Irradiation markedly induced the expression of LC3 and conversion of LC3-I to LC3-II by irradiated TE-1 cells. Compared with radiation alone and CQ pre-treatment, post-irradiation with addition of CQ significantly reduced accumulation and fluorescence intensity of AVO in surviving progenies of irradiated cells, and clonogenic survival of TE-1 cells and pre-irradiated TE-1 cells and their corresponding D0 (Gy), Dq (Gy) and SERD0 were 1.39, 0.20, 1.44 and 1.15, 0.31, 2.39, respectively. Likewise, pretreatment of pre-irradiated TE-1 cells with CQ significantly reduced clonogenic survival of irradiated cells and their corresponding D0 (Gy), Dq (Gy) and SERD0 were 1.78, 0.24, 1.54, respectively. Conclusions: Our findings suggest that blocking of autophagy would make esophageal cancer cells more sensitive to ionizing radiation and application of CQ could be used as an adjuvant for improving radiation therapy when radio-resistance develops as a result of autophagy response. Author Disclosure: X. Yuan: None. J. Huang: None. X. Zha: None. B. Chen: None. B. Deng: None. T. Li: None.

Enhancement of Radiosensitivity and the Potential Mechanism on Human Esophageal Carcinoma Cells by Tetrandrine

The purposes of this study were to test the effect of tetrandrine, alone or combined with radiation, on human esophageal cancer cell line TE1 (TE1 cells) and investigate the potential antitumor mechanism. Human esophageal cancer cell line TE1 was tested by methyl thiazolyl tetrazolium assay for cell proliferation, colony-forming assay for cell radiosensitivity, flow cytometry assay for cell cycle distribution, and western blot assay for cell cycle protein expression. When treated alone, tetrandrine had a time- and concentration-dependent cytotoxic effect on TE1 cells. The dose-enhancement ratio for combined tetrandrine and radiation was markedly increased when compared with tetrandrine alone. Further, expression of cyclin B1 protein increased after addition of tetrandrine when compared with radiation only. Radiation-induced G2 arrest was abrogated with treatment of tetrandrine. In conclusion, tetrandrine can enhance the radiosensitivity of TE1 cells and this may involve relief of radiation-induced G2/M arrest in TE1 cells.

VEGF-C promotes the development of esophageal cancer via regulating CNTN-1 expression

Vascular endothelial growth factor C (VEGF-C) is a key regulator of angiogenesis and lymphangiogenesis. VEGF-C is also implicated in the development of esophageal cancer. We investigated the mRNA levels of VEGF-C and its receptors in 38 esophageal squamous cell carcinoma specimens (ESCCs) and matched adjacent normal esophageal tissues via real-time PCR. The mRNA levels of VEGF-C, VEGFR-2 and VEGFR-3 were significantly upregulated in ESCCs versus respective side normal tissues. To explore the influence of VEGF-C on esophageal cancer progression, the expression of VEGF-C was manipulated in esophageal cancer cell lines TE-1 and Eca-109. VEGF-C transcription, translation and secretion were significantly enhanced in cells stably transfected with a VEGF-C overexpression vector or attenuated in VEGF-C shRNA-transfected cell lines. In vitro, TE-1 cells stably transfected with a VEGF-C overexpression vector exhibited an increased rate of cell proliferation, migration and focus formation, whereas knockdown of VEGF-C inhibited cell proliferation, migration and focus formation. Similar results were obtained for Eca-109 cells. VEGF-C mediated biological function through transcription of CNTN-1, which is implicated in tumor invasion and metastasis. The expression of VEGF-C was correlated with that of CNTN-1 and cell proliferation and migration induced by VEGF-C were reversed by silencing of CNTN-1. In addition, nude mice inoculated with VEGF-C shRNA-transfected cells exhibited a significantly decreased tumor size in vivo via reduced VEGFR-2 and VEGFR-3 phosphorylation and microvessel formation. VEGF-C upregulation may be involved in esophageal tumor progression. Vector-based RNA interference (RNAi) targeting VEGF-C is a potential therapeutic method for human esophageal carcinoma.

Restoration of wild-type conformation and activity of a temperature-sensitive mutant of p53 (p53(V272M)) by the cytoprotective aminothiol WR1065 in the esophageal cancer cell line TE-1.

The aminothiol WR1065, the active metabolite of the cytoprotector amifostine, exerts its antimutagenic effects through free-radical scavenging and other unknown mechanisms. In an earlier report, we showed that WR1065 activates wild-type p53 in MCF-7 cells, leading to p53-dependent arrest in the G(1) phase of the cell cycle. To determine whether WR1065 activates p53 by modulating protein conformation, we analyzed its effects on p53 conformation and activity in the esophageal cancer cell line TE-1. This cell line contains a mutation in codon 272 of p53 (p53(V272M), with methionine instead of a valine), conferring temperature-sensitive properties to the p53 protein. At the nonpermissive temperature (37 degrees C), p53(V272M) adopts the mutant p53 conformation (nonreactive with the antibody PAb1620), does not bind specifically to DNA, and is not activated in response to DNA-damaging treatment. However, treatment with 0.5-4 mM WR1065 partially restored wild-type conformation at 37 degrees C, stimulated DNA binding activity, and increased the expression of p53 target genes WAF-1, GADD45, and MDM2, leading to cell-cycle arrest in G(1). These results suggest that WR1065 activates p53 through a mechanism distinct from DNA-damage signaling, which involves modulation of p53 protein conformation.