Pyr3 Induces Apoptosis and Inhibits Migration in Human Glioblastoma Cells

Background/Aims: Glioblastoma, also known as glioblastoma multiforme (GBM), is a fast-growing type of tumor that is the most aggressive brain malignancy in adults. According to GEO profile analysis, patients with high transient receptor potential canonical 3 (TRPC3) expression have poor survival rates. The aim of this study is to evaluate the effects of Ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate (Pyr3), a selective TRPC3 channel blocker, on the proliferation and migration of human glioblastoma cells. Methods: We first analyzed the TRPC3 mRNA expression in Gene Expression Omnibus (GEO) database. Then, TRPC3 protein expression was analyzed by Western blotting in three human GBM cell lines. The survival rate was measured by sulforhodamine B. JC1 staining was used to analyze the mitochondria membrane potential by flow cytometric analysis. Besides, the migration and invasion were evaluated by wound healing and Transwell assays. Annexin V and 7-aminoactinomycin D staining was used to monitor the apoptosis by flow cytometric analysis. The expression of apoptotic-related and migration-related proteins after Pyr3 treatment was detected by Western blotting. In addition, an orthotropic xenograft mouse model was used to assay the effect of Pyr3 in the in vivo study. Results: Basis on the results of bioinformatics study, glioma patients with higher TRPC3 expression had a shorter survival time than those with lower TRPC3 expression. GBM cell proliferation was decreased by Pyr3 treatment. The migration and invasion abilities of glioma cells were also inhibited via focal adhesion kinase and myosin light chain dephosphorization after Pyr3 treatment. Moreover, Pyr3 induced caspase-dependent apoptosis and mitochondria membrane potential imbalance in the GBM cells. In a xenograft animal model, Pyr3 in combination with temozolomide (TMZ) inhibited GBM tumor growth. Conclusion: Pyr3 inhibited GBM tumor growth in vitro and in vivo. Pyr3-TMZ combination therapy could be used to treat glioblastoma in the future

Lymphovascular space invasion and tumor differentiation are predictors for postoperative recurrence in patients with pathological stage I nonsmall cell lung cancer

Abstract Background: We investigated factors predicting postoperative recurrence in patients with pathological Stage I nonsmall cell lung cancer (NSCLC). Methods: All patients with clinical Stage I NSCLC who underwent surgical resection at Tri-Service General Hospital in Taiwan between January 2002 and June 2006 were reviewed retrospectively. All study patients underwent standard staging workups. We reviewed the records of 261 patients with an average follow-up of 93 months; we then included 179 patients with pathological Stage I. Results: Two hundred sixty-one patients with clinical Stage I NSCLC were eligible. There were no significant differences in sex, tumor histopathology, location, and age between the two groups (recurrence and nonrecurrence), except for tumor differentiation (p ¼ 0.002), survival rate (p < 0.001), lymphovascular space invasion (LVSI; p ¼ 0.007), advanced pathology stage (p ¼ 0.022), maximum standard uptake value (SUVmax; p ¼ 0.027), tumor size (p < 0.011), and carcinoembryonic antigen (CEA) levels (p ¼ 0.013). Overall survival was significantly related to postoperative recurrence (p < 0.001) in patients with pathological Stage I, in whom recurrences developed in 11.17%. Only 179 patients with pathological Stage I NSCLC, including 20 patients with postoperative recurrences, were selected. Tumor differentiation (odds ratio 3.581, p ¼ 0.058) and LVSI (odds ratio 5.374, p ¼ 0.020) were independent factors predicting recurrence. Conclusion: Tumor differentiation and LVSI were predictors of postoperative relapse for patients with pathological stage I NSCLC. Risk factors of postoperative recurrence in patients with pathological Stage I NSCLC may enable us to optimize the patient selection for postoperative adjuvant therapies to prevent possibly occult micrometastases

Nrf2 Expressions Correlate with WHO Grades in Gliomas and Meningiomas

Background: Nuclear factor erythroid 2-related factor 2 (NFE2L2, also known as Nrf2) is associated with cellular progression and chemotherapeutic resistance in some human cancers. We tested the relationship between Nrf2 expression and survival of patients with primary brain tumors (PBTs). Methods: In order to realize Nrf2 protein expression in gliomas, Western blot analysis was performed in normal brain tissue and U87MG, LN229, GBM8401 and U118MG glioma cell lines protein lysates. Then, U87MG, LN229, and GBM8401 mRNA were applied to performed quantitative RT-PCR for detect Nrf2 gene expression in glioma cell lines. At last, immunohistochemical analysis was used to determine the expression of Nrf2 in samples from 178 PBTs and 10 non-neoplastic brain tissues. Results: In these included in vitro studies, both Nrf2 protein and mRNA expression in all human glioma cell lines were higher than normal brain tissue. Similarly, on the viewpoint of immunohistochemistry, Nrf2 expression in gliomas were positively correlated with World Health Organization (WHO) grades. Additionally, compared with the expression of Nrf2 in non-neoplastic brain tissue, expression in meningiomas was of a stronger intensity and was present in a higher percentage of cells. Furthermore, scores were significantly higher in WHO grade II than in WHO grade I meningiomas. Finally, overall survival tended to be shorter in patients whose PBTs had higher expression of Nrf2, although the correlation was not statistically significant. Conclusions: Nrf2 overexpression positively correlated with WHO grade in gliomas and meningiomas. On the other hand, Nrf2 immunohistochemical stain could help pathologists to differentiate atypical meningiomas from benign tumors. Therefore, Nrf2 expression may be a useful biomarker to predict WHO grade and cellular behavior of PBTs

KDELC2 Upregulates Glioblastoma Angiogenesis via Reactive Oxygen Species Activation and Tumor-Associated Macrophage Proliferation

Glioblastoma is notorious for its rapid progression and neovascularization. In this study, it was found that KDEL (Lys-Asp-Glu-Leu) containing 2 (KDELC2) stimulated vasculogenic factor expression and induced human umbilical vein endothelial cell (HUVEC) proliferation. The NLRP3 inflammasome and autophagy activation via hypoxic inducible factor 1 alpha (HIF-1α) and mitochondrial reactive oxygen species (ROS) production was also confirmed. The application of the NLRP3 inflammasome inhibitor MCC950 and autophagy inhibitor 3-methyladenine (3-MA) indicated that the above phenomenon activation correlated with an endothelial overgrowth. Furthermore, KDELC2 suppression decreased the endoplasmic reticulum (ER) stress factors’ expression. The ER stress inhibitors, such as salubrinal and GSK2606414, significantly suppressed HUVEC proliferation, indicating that ER stress promotes glioblastoma vascularization. Finally, shKDELC2 glioblastoma-conditioned medium (CM) stimulated TAM polarization and induced THP-1 cells to transform into M1 macrophages. In contrast, THP-1 cells co-cultured with compensatory overexpressed (OE)-KDELC2 glioblastoma cells increased IL-10 secretion, a biomarker of M2 macrophages. HUVECs co-cultured with shKDELC2 glioblastoma-polarized THP-1 cells were less proliferative, demonstrating that KDELC2 promotes angiogenesis. Mito-TEMPO and MCC950 increased caspase-1p20 and IL-1β expression in THP-1 macrophages, indicating that mitochondrial ROS and autophagy could also interrupt THP-1-M1 macrophage polarization. In conclusion, mitochondrial ROS, ER stress, and the TAMs resulting from OE-KDELC2 glioblastoma cells play important roles in upregulating glioblastoma angiogenesis

Proteolipid Protein 2 Overexpression Indicates Aggressive Tumor Behavior and Adverse Prognosis in Human Gliomas

Proteolipid protein 2 (PLP2), a membrane protein of the endoplasmic reticulum, is related to tumor proliferation and metastasis in some human cancers, but not in gliomas. First, we performed western-blot analysis, real-time quantitative PCR and immunohistochemical stains to detect PLP2 expression in 4 glioma cell lines and human glioma tissues. In addition, we used small interfering RNA (SiPLP2) and short hairpin RNA (shPLP2) to knockdown PLP2 expression in GBM8401 and LN229 glioma cell lines. After then, the alteration of PLP2 suppressed glioma cells behavior were examined by cell proliferation, wound healing, cell invasion, and colonies formation assays. Finally, the possible mechanism of PLP2 was analyzed by detecting the expression of the proteins related to cell-cycle checkpoints, cell-proliferative signaling factors, and cell-matrix interaction. Compared with normal brain cell lysates and mRNA, all glioma cell lines displayed PLP2 protein and mRNA overexpression. Besides, higher PLP2 IHC staining significantly correlated with more advanced tumor grades and poorer prognosis in human gliomas. Both siPLP2 transfected gliomas showed a clear inhibition of glioma cell proliferation, migration, and invasion as well as down-regulating p-p38, p-ERK, MMP-2, and MMP-9 expression. In conclusion, we successfully demonstrated that PLP2 overexpression played an oncogenic role in glioma development and aggressive tumor behavior

Hepatic cystic metastatic tumors from a locally controlled nasopharyngeal carcinoma

Liver cystic neoplasms are uncommon and vary from benign to overtly malignant. Liver cystic metastases are rare and mostly come from colon, pancreas, ovary, kidney, neuroendocrine, and prostate cancer. Nasopharyngeal carcinoma (NPC) with liver cystic metastasis has only been reported once. Here, we report a 52-year-old man with liver cystic metastasis from locally cured NPC. The patient received concurrent chemoradiotherapy for NPC 4 years ago and presented with a 6-month history of upper abdominal fullness and pain. No evidence of local recurrence of NPC was found at his regular follow-up examinations after concurrent chemoradiotherapy. Abdominal magnetic resonance imaging showed a large, well-defined, lobulated cystic lesion with poor contrast enhancement occupying both lobes of the liver. Hepatic cystic metastasis was suspected. Ultrasound-guided liver tumor biopsy was performed. Histological examinations disclosed a pattern of poorly differentiated squamous cell carcinoma with focal sarcomatoid differentiation based on the P40 immunohistochemical stain. In situ hybridization for Epstein–Barr virus early RNAs confirmed the diagnosis of metastatic NPC. It is difficult to make a diagnosis in liver cystic neoplasms, especially from a rarely reported origin. In our case, we used clinical history and Epstein–Barr virus early RNAs as a specific marker to make an accurate diagnosis