Squamous Carcinoma Cell Line Eca109 Research Articles

Bortezomib sensitizes esophageal squamous cancer cells to radiotherapy by suppressing the expression of HIF-1α and apoptosis proteins.

Radiation therapy is a typical treatment for esophageal squamous cell carcinoma (ESCC), especially middle and upper segment esophagus, and inoperable patients. However, how to promote radiation sensitivity in radio-resistant cancer cells is a conundrum. Here, our study investigated the radiosensitizing effect of bortezomib, a specific and reversible dipeptide boronic acid analog, in ESCC cells. Human esophageal squamous carcinoma cell lines Eca109 and TE-13 were exposed to hypoxia and/or ionizing radiation (IR) with or without treatment of bortezomib. Cell proliferation assay was performed with CCK8. Cell apoptosis and cell cycle assay were performed with flow cytometry. The radiosensitization effect of was assessed by clonogenic survival and progression of tumor xenograft. The expression of HIF-1α, VEGF, and apoptosis proteins was evaluated by Western blot. Radiation-induced DNA double strand break and homologous recombination repair were assessed by immunofluorescence. Our results show that bortezomib efficiently radiosensitizes ESCC cells by decreasing the expression of HIF- 1α and VEGF, inducing apoptosis by activating caspase, and delaying DNA damage repair after radiation.

157P - Contribution of Mesenchymal Stem Cells (Mscs) on Epithelial-Mesenchymal Transition (Emt) of Esophageal Squamous Cell Carcinoma

ABSTRACT Aim: Mesenchymal stem cells (MSCs) were recently found to be one of the candidates involved in the development of tumor stroma. Emerging evidence links MSCs in the tumor microenvironment with the occurrence of epithelial-mesenchymal transition (EMT) in cancer progression. Here we report that MSCs may contribute to inducing EMT of esophageal squamous carcinoma cells. Methods: The human esophageal squamous carcinoma cell lines Eca-109 and TE-1 were cultured with supernate of human MSCs. The cultured tumor cells were analyzed for changes in cellular morphology, EMT markers, protein expression and tumor characteristics by immunofluorescence analysis, q-PCR, and Western-Blot. Invasiveness of MSC-conditioned tumor cells was also observed in a transwell system. Results: We found that MSC-conditioned esophageal squamous carcinoma cells underwent EMT and established a stable mesenchymal phenotype after prolonged culturing, as indicated by upregulation of EMT-related genes, downregulation of E-cadherin, and acquisition of mesenchymal morphology. Different concentrations of MSC-derived conditioned medium would induce similar changing of tumor cells. Furthermore, MSC-conditioned esophageal squamous carcinoma cells displayed increased invasiveness in a transwell system. Conclusions: By promoting EMT-related phenomena, MSC appear to favor the improvement of aggressiveness and invasiveness of esophageal squamous carcinoma cells. Thus, MSCs may contribute to inducing EMT of esophageal squamous carcinoma cells. Disclosure: All authors have declared no conflicts of interest.

Effect of miR-214 on invasive capacity of esophageal squamous carcinoma cell line Eca109

Objective To investigate the effect of miR-214 on the invasive capacity of esophageal squamous carcinoma cell line Eca109,and to explore the possible molecular mechanism.Methods We prepared miR-214 double-stranded mimic and transfected it into Eca109 cells with Lipofectamine 2000.Eca109 cells transfected with nonsense miRNA mimics were taken as control.The expression of mature miR-214 was determined by qPCR.The capacity of cell invasion was determined by Matrigel-coated Transwell assay.E-cadherin protein expression and the percentage of E-cadherin positive cells were examined by Western blotting analysis and flow cytometry,respectively.Results The expression level of mature miR-214 in the miR-214 mimic transfection group was significantly higher than that in the control group 48 h after transfection(P0.01).Eca109 cells transfected with miR-214 mimic showed a significantly lower cell invasive capacity compared to that of cells transfected with control miRNA mimic(P0.05).Moreover,E-cadherin protein expression and the ratio of E-cadherin positive cells in miR-214 mimic transfection group were both significantly lower than those in the control group(P0.05).Conclusion Our data suggest that miR-214 may inhibit the invasive capacity of esophageal squamous carcinoma cells by repressing the epithelial-mesenchymal transition.

Luteolin Induced-growth Inhibition and Apoptosis of Human Esophageal Squamous Carcinoma Cell Line Eca109 Cells in vitro

Luteolin is a plant flavonoid which exhibits anti-oxidative, anti-inflammatory and anti-tumor effects. However, the antiproliferative potential of luteolin is not fully understood. In this study, we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells. MTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an IC50 of 70.7±1.72 μM at 24 h. Luteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM. Luteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining. Moreover, luteolin downregulated the expression of cyclin D1, survivin and c-myc, and it also upregulated the expression of p53, in line with the fact that luteolin was able to inhibit Eca109 cell proliferation.

Cyclic bisdesmosides from Actinostemma lobatum MAXIM (Cucurbitaceae) and their in vitro cytotoxicity

Two new cyclic bisdesmosides elucidated as lobatoside L ( 1) and lobatoside M ( 2) and four known cyclic bisdesmosides ( 3– 6) were isolated from Actinostemma lobatum MAXIM (Cucurbitaceae). Their structures were determined on the basis of spectroscopic analyses including IR, HR-FAB-MS, TOCSY, 1D- and 2D-NMR experiments and chemical reactions. The inhibitory effects of the six compounds on human cancer cell growth (including esophageal squamous carcinoma cell line ECA109, lung cancer cell line A549 and gastric cancer cell line MGC-803) were determined using the MTT assay. The results revealed that the six compounds exhibited cytotoxicity against all the cell lines tested, and compounds 1, 3 and 5 showed significant activities in a dose dependent manner against all the cell lines, especially for compound 1 and 5, the IC 50 values for ECA-109 cells were 8.25 μM and 3.71 μM. The results demonstrated that compounds 1 and 5's activities are 2- and 4-fold that of cisplatin.

Effects of YC-1 targeting hypoxia-inducible factor 1 alpha in oesophageal squamous carcinoma cell line Eca109 cells

HIF-1α (hypoxia-inducible factor 1 alpha) is believed to promote oesophageal squamous tumour growth. Thus, an HIF-1α inhibitor is viewed as a therapeutic target in treating oesophageal cancer. Recently, YC-1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole] has been widely used as a potential HIF-1α inhibitor and is being developed as a novel anticancer drug. However, little is known about the effects of YC-1 in human oesophageal cancer. In the present study, we aimed to investigate these effects in an esophageal squamous cancer cell line; i.e. Eca109 cells. We found that YC-1 abolished the hypoxia-induced up-regulation of HIF-1α. YC-1 arrested cell growth and inhibited cell migration activities in Eca109 cells. These results suggest that YC-1 may be a chemotherapy candidate against oesophageal squamous cancers.

Effect and mechanism of 5-aminolevulinic acid-mediated photodynamic therapy in esophageal cancer

5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) provides a novel and promising treatment for esophageal cancer. However, its specific mechanism has not been fully elucidated and its efficacy is remarkably varied. This study investigated the effect of ALA-PDT on esophageal squamous carcinoma cell line Eca-109 in vitro and vivo to explore optimal parameters, and evaluated the significance of cell apoptosis, cell cycle, ALA-protoporphyrin IX (ALA-PpIX) subcellular localization, and expression of Bcl-2 and Bax mRNA in cells to understand the mechanism of ALA-PDT for esophageal cancer. How ALA concentration, incubation time, and laser irradiation dose influenced the cell proliferation was determined by MTT assay. ALA-PpIX subcellular localization was analyzed by confocal microscopy. The mRNA changes were detected by quantitative real-time polymerase chain reaction (QRT-PCR). Tumor models transplanted with Eca-109 cells in nude mice were established (n = 10) and killed (n = 4) at 24 h post-PDT for malondialdehyde (MDA) detection and histological study. The remaining mice were measured the tumor size for 3 weeks after treatment. Our data show that ALA-PDT significantly inhibits cell proliferation (p < 0.05), the PDT efficacy depends on the saturation of ALA concentration, incubation time, and laser irradiation dose, and the best effect in tumor destruction is at 7-14 days post-PDT. ALA-PpIX is localized in mitochondria and cytoplasm. ALA-PDT induces cell apoptosis and arrests cell cycle at G0/G1 phase. Bcl-2 is significantly down-regulated while Bax is up-regulated (p < 0.05). The results of this study provide references in choosing clinical optimal PDT parameters and help in better understanding the PDT mechanism for esophageal cancer.

Inhibitory effect of recombinant LIGHT-Fc gene on the proliferation of human esophageal squamous carcinoma cell line Eca109

Objective To investigate the inhibitory effect of recombinant LIGHT-Fc gene on the proliferation of human esophageal carcinoma cell line Eca109. Methods LIGHT-Fc expression vector was transfected into human esophageal squamous carcinoma cell line Eca109 by using DOTAP liposomal transfection reagents. The effects of LIGHT-Fc gene on the proliferation of esophageal carcinoma cell line Eca109 in vitro were detected by cell growth curve and MTT assay, Forty-five nude mice were equally divided into Eca109/Wt group, Eca109/neo group and Eca109/LIGHT group. Carcinogenesis and pathological expression of the esophageal carcinoma tissues were observed. Results The expressions of LIGHT receptors were detected in Eca109 cells. The proliferation of Eca109 cells was inhibited after trasfecting LIGHT-Fc gene into Eca109 cells. The numbers of tumors generated in Eca109/Wt group, Eca109/neo group and Eca109/LIGHT group were 12, 11 and 5, with statistical significance between Eca109/LIGHT group and the other two groups （ X^2 = 6. 652, 4. 821, P 〈 0.05 ）. The result of histopathological examination indicated that the tissue necrosis appeared significantly in tumors derived from Eca109/LIGHT cells. Conclusions The growth of esophageal squamous carcinoma cell line Eca109 can be suppressed by LIGHT-Fc gene whether in vitro or in vivo. Key words: Recombinant gene ; Transfection ; Esophageal carcinoma

The flavonoid component isorhamnetin in vitro inhibits proliferation and induces apoptosis in Eca-109 cells

Isorhamnetin is one member of flavonoid components which has been used in the treatment of heart disease. Recently the in vitro anti-cancer effect of isorhamnetin on human esophageal squamous carcinoma cell line Eca-109 was investigated in our lab. When Eca-109 cells were in vitro exposed to the graded doses of isorhamnetin (0–80 μg/ml) for 48 h, respectively, isorhamnetin exhibited cytostatic effect on the treated cells, with an IC 50 of 40 ± 0.08 μg/ml as estimated by MTT assay. Inhibition on proliferation by isorhamnetin was detected by trypan blue exclusion assay, clone formation test, immunocytochemical assay of PCNA and 3H-thymidin uptake analysis. Cell cycle distribution was measured by FCM. It was found that the viability of Eca-109 cells was significantly hampered by isorhamnetin. Compared with the negative control group, the treated group which was exposed to isorhamnetin had increased population in G 0/G 1 phase from 74.6 to 84 while had a significant reduction in G 2/M phase from 11.9 to 5.8. In addition to its cytostatic effect, isorhamnetin also showed stimulatory effect on apoptosis. Typical apoptotic morphology such as condensation and fragmentation of nuclei and blebbing membrane of the apoptotic cells could be observed through transmission electron microscope. Moreover, the sharp increase in apoptosis rate between the control and treated group were detected by FCM from 6.3 to 16.3. To explore the possible molecular mechanisms that underlie the growth inhibition and apoptosis stimulatory effects of isorhamnetin, the expressions of six proliferation- and death-related genes were detected by FCM. Expressions of bcl-2, c-myc and H-ras were downregulated whereas Bax, c-fos and p53 were upregulated. However, the in vivo experiments were required to further confirm the anti-cancer effects of isorhamnetin. In conclusion, isorhamnetin appears to be a potent drug against esophageal cancer due to its in vitro potential to not only inhibit proliferation but also induce apoptosis of Eca-109 cells.