ABSTRACT
Triptolide, the component of traditional Chinese herb, has been used as an inflammatory medicine and reported to be anti-tumor for various cancers recently. However, the effect of triptolide on Esophageal Squamous Cell Cancer (ESCC) has not yet been elucidated. In the study, we found that triptolide significantly inhibited cell proliferation, invasion, migration and survivability of ESCC cells. Moreover, we observed that triptolide induced ESCC cell cycle arrest at the G1/S phase and apoptosis through cyclin D1-CDK4/6 regulation and caspases activation. In addition, we revealed that triptolide regulates cell apoptosis and metastasis by p53 and mitogen-activated protein kinases/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway, respectively. Meanwhile, the inhibitory effect of triptolide on ESCC was validated in mouse xenograft model. So, we propose that triptolide may be a candidate drug for ESCC.
Subject(s)
Cell Movement/drug effects , Diterpenes/pharmacology , Esophageal Neoplasms/pathology , Esophageal Squamous Cell Carcinoma/pathology , Extracellular Signal-Regulated MAP Kinases/metabolism , MAP Kinase Signaling System/drug effects , Mitogen-Activated Protein Kinases/metabolism , Phenanthrenes/pharmacology , Animals , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Epoxy Compounds/pharmacology , Humans , Mice , Neoplasm Metastasis , Xenograft Model Antitumor AssaysABSTRACT
Objective To investigate the effects of methylation inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC) on biological behavior of esophageal squamous carcinoma cell (ESCC) lines KYSE140 and KYSE150. Methods KYSE140 and KYSE150 cell lines were divided into the blank group, the control group and the experimental group. The cells in the blank group didn't do the treatment, and the cells in the control group were added to DMSO 2 μmol/L, while in the experimental group, cells were treated with different concentration (1, 2, 3 and 4 μmol/L) of 5-Aza-dC which affected respectively at different time (24, 48, 72 and 96 h). Cell proliferation was detected by using methyl thiazolyl tetrazolium (MTT) assay and the optimal drug concentration and time point were selected. Transwell assay was performed to detect the change of cell migration and invasion. Flow cytometry was used to observe the effects of drugs on cell apoptosis and cell cycle.The expression of PARP,Caspase-3,CCNB-1,and CCNE-1 were detected by Western blot. Results MTT result showed that the effective function time of 5-Aza-dC on KYSE140 and KYSE150 was 96 h at the concentration of 4 μmol/L. Under this condition, the cell ability of migration and invasion was decreased significantly. The migrated cell number of KYSE140 and KYSE150 respectively in the blank group, the control group and the experimental group was (193.3±8.6), (184.0±10.4), (61.7±7.1) and (112.0±6.4), (101.3± 7.9), (26.3±5.7). The invasive cell number was (47.3±7.3), (38.7±5.1), (8.0±3.9) and (83.3±6.8), (74.7±5.7), (21.0±2.7), respectively. The difference was statistically significant (P <0.05). Flow cytometry revealed that 5-Aza-dC increased the apoptosis of KYSE140 and KYSE150. The apoptosis rate of the blank group, the control group and the experimental group was (2.8±0.3) %, (11.2±0.7) %, (18.6±0.6) % for KYSE140 and (2.7±0.4)%,(9.8±0.4)%,(17.7±0.5)% for KYSE150.Compared with the other two groups,the cell number of G2/M phase in the experimental group was increased remarkably (P < 0.05). PARP and Caspase-3 were sheared evidently and the protein expression of CCNB-1 was up-regulated while the expression of CCNE-1 was down-regulated in the experimental group. Conclusion 5-Aza-dC can inhibit the proliferation and promote apoptosis of ESCC cells.
ABSTRACT
Objective:To analyze the expression of LETM2 in KYSE150 and ECA109 cell lines and its effect on the proliferation, migra-tion, and invasion of esophageal squamous cell carcinoma (ESCC). Methods:The expression level of the LETM2 protein in 90 paired hu-man ESCC tissues and matched adjacent normal tissues was determined through immunohistochemistry. The expression level of LETM2 in ESCC cell lines was detected by real-time PCR and Western blot. The expression levels of LETM2 in KYSE150 and ECA109 cell lines were knocked down using lentivirus. MTT assays were performed to examine the effect of LETM2 on the proliferation of ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle. The effect of LETM2 depletion on the migration and invasion of ESCC cells was determined by Transwell assay. Results:LETM2 expres-sion was frequently upregulated in the ESCC tissues than in the adjacent normal tissues. The suppressed exogenous expression of LETM2 led to the inhibition of cell proliferation and colony formation. However, cell migration and invasion were not affected. The re-sults on the cell cycle distribution revealed that LETM2 knockdown acts as a negative regulator of the cell cycle at the G1 to S phase transition. Conclusion:LETM2 acts as a tumor-driven gene in the development and progression of ESCC. This finding suggests that LETM2 can be used as an efficient prognosis biomarker and a potential therapeutic target for ESCC.
ABSTRACT
Objective:To analyze the expression of MUC15 and PI3K/Akt in gastric carcinoma and its association with clinicopathologi-cal characteristics and prognosis. Methods:The expression of MUC15 and Akt was detected in 144 cases of gastric carcinoma tissues and corresponding para-carcinoma tissues by tissue microarray and immunohistochemistry. Results:The positive expression rate of MUC15 in gastric carcinoma was 79.8%, higher than that of para-carcinoma tissues (22.2%, P<0.01). The positive expression rate of Akt protein in gastric carcinoma was 80.6%, higher than that of para-carcinoma tissues (16.7%, P<0.01). The expression of MUC15 and Akt was statistically associated with the grades of differentiation, invasion depth, lymphatic metastasis, TNM stage of tumor tissues (P<0.05), and the positive correlation between the two protein expression that appear in the gastric tumor tissue (P=0.001). Univariate survival analysis showed that the over-expression of either MUC15 or Akt was inversely correlated with the survival time (P<0.05 and P<0.01, respectively). Cox multiple regression analysis indicated that patients with over-expression of both MUC15 and Akt had the worst prognoses (HR=3.115, P<0.05). Conclusion:MUC15 may be involved in the occurrence, development, invasion, and metastasis of gastric cancer through the PI3K/Akt cell signaling pathway, and the expression of MUC15 combined with Akt is a powerful predictor for the prognosis of gastric cancer.
