alpha-asarone and Glioma

Glioma is the most common type of primary brain tumor and has an undesirable prognosis. Autophagy plays an important role in cancer therapy, but it is effect is still not definite. P53 is an important tumor suppressor gene and protein that is closely to autophagy. Our aim was to study the effect of β-asarone on inhibiting cell proliferation in human glioma U251 cells and to detect the effect of the inhibition on autophagy through the P53 signal pathway. For cell growth, the cells were divided into four groups: the model, β-asarone, temozolomide (TMZ), and co-administration groups. For cell autoghapy and the P53 pathway, the cells were divided into six groups: the model, β-asarone, 3MA, Rapa, Pifithrin-µ, and NSC groups. The counting Kit-8 assay and flow cytometry (FCM) were then used to measure the cell proliferation and cycle. Electron microscopy was used to observe autophagosome formation. Cell immunohistochemistry/-immunofluorescence, FCM and Western blot (WB) were used to examine the expression of Beclin-1 and P53. The levels of P53 and GAPDH mRNA were detected by RT-PCR. Using WB, we determined autophagy-related proteins Beclin-1, LC3-II/I, and P62 and those of the P53 pathway-related proteins P53, Bcl-2, mTOR, P-mTOR, AMPK, P-AMPK, and GAPDH. We got the results that β-asarone changed the cellular morphology, inhibited cell proliferation, and enhanced the expression of P53, LC3-II/I, Beclin-1, AMPK, and pAMPK while inhibiting the expression of P62, Bcl-2, mTOR, and pmTOR. All the data suggested that β-asarone could reduce the cell proliferation and promote autophagy possible via the P53 pathway in U251 cells.

Topics: Allylbenzene Derivatives; AMP-Activated Protein Kinases; Anisoles; Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Autophagosomes; Autophagy; Beclin-1; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Shape; Dacarbazine; Glioma; Humans; Phosphorylation; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Temozolomide; TOR Serine-Threonine Kinases; Tumor Suppressor Protein p53

Topics: Allylbenzene Derivatives; Anisoles; Antineoplastic Agents; Brain Neoplasms; Cell Adhesion; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Survival; Dose-Response Relationship, Drug; Down-Regulation; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Glioma; Heterogeneous-Nuclear Ribonucleoproteins; Humans; Neoplasm Invasiveness

Topics: Allylbenzene Derivatives; Animals; Anisoles; Apoptosis; Brain Neoplasms; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cell Survival; Disease Models, Animal; Glioma; Heterogeneous-Nuclear Ribonucleoprotein Group A-B; Humans; Mice; Mitochondria; Proto-Oncogene Proteins c-bcl-2; RNA Splicing; Signal Transduction; Xenograft Model Antitumor Assays

Topics: Allylbenzene Derivatives; Animals; Anisoles; Antineoplastic Agents; Apoptosis; Apoptosis Regulatory Proteins; Brain Neoplasms; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Movement; Cell Proliferation; Dose-Response Relationship, Drug; Glial Fibrillary Acidic Protein; Glioma; Humans; Inhibitory Concentration 50; Neoplasm Invasiveness; Neuropilin-1; Phosphopyruvate Hydratase; Rats; Signal Transduction; Time Factors

Glioma is the most common primary brain tumor and has an undesirable prognosis due to the blood-brain barrier (BBB) and drug resistance. A thorough investigation of the changes in intracellular drug concentrations is important to observe therapeutic effects and cell resistance. P-glycoprotein (P-gp) is an essential protein of Multi-drug resistance 1 (MDR1). The over-expression of P-gp and MDR1 is associated with poor prognosis and drug-resistance in glioma. However, β-asarone can pass through the BBB easily and increase the drug concentration in the rat brain. Our aim is to study the effect of β-asarone on promoting the entry of temozolomide (TMZ) into human glioma U251 cells. The cells were divided into three groups: model group, TMZ group (300μM) and co-administration group (360μM β-asarone; 300μM TMZ). We further detected P-gp and MDR1 expression in U251 and rat glioma C6 cells in four groups: model group (U251/C6), TMZ group (U251 300μM, C6 420μM), β-asarone group (U251 360μM, C6 450μM) and co-administration group (β-asarone 360μM, TMZ 300μM for U251; β-asarone 450μM, TMZ 420μM for C6). Then, high performance liquid chromatography was used to determine the intracellular and extracellular levels of TMZ. Morphological changes in both cells were observed by the microscope. The Counting Kit-8 assay was used to measure the cell proliferation and toxicity. Cell immunohistochemistry/immunofluorescence, flowcytometry and western blot were synchronously used to examine the expression of P-gp. We also determined the levels of MDR1 mRNA by RT-PCR. The results showed that β-asarone could promote the entry of TMZ into U251 cells through the membrane. The co-administration of β-asarone and TMZ also decreased cell proliferation and the expression of P-gp and MDR1 better than single medication in U251 and C6 cells. All of the data suggest that β-asarone might contribute to treatment by promoting TMZ's entry into glioma cells, thereby contributing to anti-cancer growth and inhibiting P-gp and MDR1 expression.

Topics: Allylbenzene Derivatives; Animals; Anisoles; Antineoplastic Agents, Alkylating; ATP Binding Cassette Transporter, Subfamily B; ATP Binding Cassette Transporter, Subfamily B, Member 1; Blood-Brain Barrier; Brain Neoplasms; Cell Line, Tumor; Cell Proliferation; Dacarbazine; Drug Resistance, Neoplasm; Glioma; Humans; Rats; RNA, Messenger; Temozolomide